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Table of Content

    05 May 2017, Volume 37 Issue 5
    Acridine orange staining for fast identification of the spermatogenic cells during STA-PUT velocity sedimentation
    2017, 37(5):  591-595. 
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    Objective To establish a quick and accurate technique of identifying the pachytene spermatocytes,round spermatids and elongating and condensing spermatids during STA-PUT velocity sedimentation. Methods Using STA-PUT velocity sedimentation method to isolate the pachytene spermatocytes, round spermatids and elongating and condensing spermatids from mouse testes.To determine the cell populations’ distribution ,each tube of cell fraction was then partially transfered to the 96 plate well ,and each well was added with acridine orange dye.Then each well was analyzed using fluorescence microscopy. Results The 3 types of spermatogenic cells can be identified quickly and accurately by it’s specific cytoplasm/nucleus character using the acridine orange dye staining under fluorescence detection. Conclusions A rubust method to quickly and accurately determine the pachytene spermatocytes,round spermatids and elongating and condensing spermatids during STA-PUT velocity sedimentation was successfully developed.
    Endotoxin induces the apoptosis of degenerated human intervertebral disc nucleus pulposus cells
    2017, 37(5):  596-600. 
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    Objective To observe the relationship between nuclear factor kappB (NF - κB) activation and cell apoptosis,before and after LPS stimulation of human degene- rateve nucleus pulposus cells. Methods Degenerative human nucleus pulposus cells were cultured in vitro.Useing CCK-8 to observe the proliferative effect of LPS on the nucleus pulposus cells in vitro, in the concentration of 100, 200, 500 and 1 000 μg/mL and choose the most apropriate concentration. The experiment was divided into 3 groups: blank control group, LPS(500 μg/mL)groups, and NF-κB inhibitor(BAY11-7082)plus LPS(500 μg/mL)group,Annexin V-FITC/PI flow cytometry and Hoechest33258 staining was used to analyze apoptosis. The expression of cleaved caspase 3,PARP,P65,P-P65 proteins were detected by Western blot respectively. Results The results of CCK-8 assay showed that when LPS was 500 μg/mL, the cell vitality was declined obviously.Compared with the blank control group, cell apoptosis rate of the LPS group is increasing(p<0.05),and the expression of P-P65,cleaved caspase 3,cleaved PARP were also higher (p< 0.05).Compared with the LPS group, cell apoptosis rate of the NF-κB inhibitor plus LPS group is significantly lower (p<0.05) and the expression of P-P65,cleaved caspase 3,cleaved PARP were also lower (p< 0.05).Conclusions NF-κB signaling pathway may be associated with the nucleus pulposus cell apoptosis in disc degeneration,so it was worth further study.
    NF-κB inhibitor reduces the expression of zinc finger protein A20 in rabbit lumbar intervertebral disc nucleus pulposus cells induced by lipopolysaccharide
    2017, 37(5):  601-607. 
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    Objective To observe the expression of zinc finger protein A20(A20), NF-κB and related inflammatory factors before and after lipopolysaccharide (LPS) stimulates degeneration of rabbit intervertebral disc nucleus pulposus cells. Methods The normal and degenerative nucleus pulposus cells were isolated and cultured in vitro,then divided into normal group,degenerative group,LPS stimulation group and NF-κB inhibition group.HE staining observe the morphological changes of nucleus pulposus and annulus fibrosus,immunohistochemistry was used to detect the expression of A20,NF-κB/p65 and COL-Ⅱ.Real-time PCR was employed to analyze the expression of A20,IL-1β,TNF-α,NF-κB and COL-Ⅱ,Western blot was used to observe the A20 protein,p65 and COL-Ⅱexpression in the four groups,and TNF-α,IL-1β in cell supernatant was determined by ELISA. Results The number of nucleus pulposus cells was significantly decreased, aggregation occurs in the degenerative group.COL-Ⅱ was obvious lower and A20, p65 was significantly higher than normal group by immunohistochemical staining. Compared with the normal group,A20,TNF-α,IL-1β,p65 expression was significantly increased and COL-Ⅱ decreased in the mRNA and protein levels in degenerative group.Above indexes change more significant in LPS stimulation group than in degenerative group.The expression of A20, TNF-α, IL-1β, p65 in the NF-κB inhibitor group was lower than that in the LPS group, and the expression of type Ⅱ collagen increased(p<0.05). Conclusions Intervertebral disc inflammatory response is closely related to the development of intervertebral disc degeneration, A20 may play an important role.
    MCPIP1 induces cell cycle arrest in breast cancer cell line MDA-MB-231
    Mingming Liu
    2017, 37(5):  608-613. 
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    Objective To investigate the functions of Monocyte chemotactic protein-induced protein 1 (MCPIP1) in human breast cancer cell line MDA-MB-231. Methods MDA-MB-231 cells were transfected with GFP-tagged MCPIP1 by Tet-on inducing expression system. Endogenous MCPIP1 was knocked down by stable expressing shRNA. MTT assay was performed to measure the growth of MDA-MB-231 cells after overexpression or knockdown of MCPIP1. FACS method was used to analysis cell cycle in MDA-MB-231 cells. Real-time PCR was used to test the expression of cell cycle-related mRNAs expression and their half-lives. RNA-IP experiment was conducted to detect the mRNA directly enriched by MCPIP1. Luciferase assay was performed to determine whether the mRNA decay was mediated through 3’UTR. Results MCPIP1 overexpression inhibited cell proliferation significantly (P<0.05), while knockdown MCPIP1 promoted cell proliferationwith statistical significances (P<0.05). MCPIP1 induced cell cycle arrest in MDA-MB-231 with statistical significance (P<0.01). MCPIP1 overexpression reduced the half-lives of cell cycle mRNAs (CDK2,CDK6,Cyclin D1,Cyclin E1,respectively) with significance (P<0.01). In addition, cell cycle-related mRNAs were able to be pulled down by GFP-MCPIP1but not isotype IgG (P< 0.05). Compared with control vector, MCPIP1 significant suppressed luciferase activitiesof all four 3’UTR reporters (P< 0.05). Conclusion MCPIP1 functions as a tumor suppressor in human breast cancer cell line MDA-MB-231 through inducing G1 cell cycle arrest.
    Effect of PI3K/Akt signaling pathway and endoplasmic reticulum stress on apoptosis of hepatocytes in rats with hepatic fibrosis
    2017, 37(5):  614-618. 
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    Objective To observe the changes of PI3K/Akt signaling pathway and endoplasmic reticulum stress related protein GRP78 and CHOP in CCl4 induced liver fibrosis and to explore their effects on hepatic fibrosis. Methods Thirty SD rats were randomly divided into normal control group, 4 and 8 weeks liver fibrosis group (hypodermic injection of 40% CCl4). Pathological changes of liver tissue was observed by HE staining. The techniques of real-time fluorescence quantitative PCR was applied to detect mRNA of GRP78 and CHOP in liver. Detected expression of Akt1, phospho-Akt1, GRP78 and CHOP protein by western blot. Meanwhile, the cell apoptosis in liver was detected by TUNEL. Results Compared with the normal control group, GRP78 and CHOP mRNA and protein in 4 and 8 weeks liver fibrosis group was increased(P<0.05), while expression of Akt1, phospho-Akt1 in 4 and 8 weeks liver fibrosis group was lower than that in normal control group(P<0.05). Compared with the normal control group, the apoptosis of hepatocytes in 4 and 8 weeks liver fibrosis group was elevated (P<0.05). Conclusions PI3K/Akt signaling pathway and endoplasmic reticulum stress may play important roles in apoptosis of hepatocytes in rats with hepatic fibrosis.
    Establishment and application of natural Chinese herbal cytological screening method targeting to PPARγ2
    2017, 37(5):  619-624. 
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    Objective To establish a natural Chinese herbal cytological screening method through the establishment of stably transfected 3T3-L1 cell line with the luciferase reporter gene expression plasmid containing human PPARγ2 gene promoter. Methods Firstly, the luciferase reporter gene expression plasmids containing various length PPARγ2 gene promoter sequence were constructed by using pGL3-Basic-luc and pGL3-Enhancer-luc as vector. The stably transfected 3T3-L1 cell line was stablished, and twenty-eight kinds of extracts of Chinese medical herbs (ECMH)were selected. Finally the effects of these ECMH on the luciferase expression in 3T3-L1 cells were observed. Results The luciferase expression of 3T3-L1 cells stably transfected withpGL3- Enhancer -PPARγ2 625bp-Luc plasmid was the highest,about 200 times than the background. Therefore, this cell line was selected for the following screening experiment. Our results showed that the administration of safflower ECMH(10,100, and 1000μg / mL)significantly promoted the luciferase expression of 3T3-L1 cells .it was 1.63 , 1.90 , and 2.30 (p<0.05) of no drug intervention cells. Similarity,Leaves and madder ECMH (10 ~ 1000μg / mL)also notably increased the luciferase expression of 3T3-L1 cells. The highest actions were observed, at a concentration of 1000μg / mL,which were 2.03 (p<0.01) and 2.00 times of the control group (p<0.01), respectively. Conclusions Stable transfection of 3T3-L1 cell line with pGL3- Enhancer -PPARγ2 625bp-Luc plasmid was successfully established. Using this cell line safflower,leaves,madder ECMH were found to significantly promote human PPARγ2 gene promoter activities, and they may become potential weight loss medicine in the future.
    Insulin resistance and low level adiponectin co-promote cardiac remodeling in mice
    2017, 37(5):  625-629. 
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    Objective The study on the effect of cardiac remodeling induced by low level adiponectin(APN)and insulin resistance(IR). Methods 16 wild-type C57 mice were randomly divided into control and IR groups.16 APN gene knockout mice were randomly divided into APNKO and APNKO+IR groups. The IR and APNKO+IR groups were a high-fat feed to induce insulin resistance.After 12 weeks,cholesterol(TC),triglycerides(TG),fasting plasma glucose(FPG), fasting insulin(FINS) were measured.HOMA-IR,HWI and LVWI were calculated.The left ventricular myocardial were dyed by Masson staining to observe the degree of myocardial fibrosis,and the protein expression of matrix metalloproteinase-9(MMP-9) and APN in myocardial were detected with western blot. Results Low level APN and IR increased heart weight, left ventricular weight, HWI, LVWI(P<0.05).There were interaction between IR and low level APN on the TC,TG, FPG, FINS, and the index related myocardial fibrosis(P<0.05), and they were positive effect. There was also interaction effect between IR and low level APN on myocardial APN expression, but it was the negative effects (P<0.05).Conclusions Both low level adiponectin and insulin resistance could lead to cardiac remodeling synergistically.
    Vincristine reduces A20 siRNA-induced proliferation in diffuse large B cell
    2017, 37(5):  630-635. 
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    Objective To investigate the impact of MDR1-targeting small interfering RNA (siRNA) on the proliferation of OCI-LY1 cells in diffuse large B-cell lymphoma. Methods A20 gene was silenced using RNA interference. An optimal concentration and treatment duration of vincristine were selected using MTT. Before and after siRNA transfection, proliferation of OCI-LY1 cells were assayed using MTT assay, and cellular apoptosis was detected using FCM before or after the treatment of the cells with VCR. Detection of A20, NF-κB (p65) and Pgp proteins were conducted using Western blotting whereas mRNA of the A20 and MDR1 genes were examined using real time PCR. Results 1. Proliferation of OCI-LY1 cells was enhanced (P<0.001) after the transfection with siRNA-2,(P<0.05). In addition, cell proliferation curve was declined after VCR stimulation, but the decrease was slower in siRNA-transfected cells than the untransfected counterparts. 2. Apoptostic rate was lower in siRNA-transfected cells than the untransfected counterparts, and the rate was higher in the cells after treatment with the drug for 24h (P< 0.05). Increased apoptosis was more obvious in common OCI-LY1 cells than in siRNA-transfected cells after the treatment with VCR(P<0.05). 3. The expression of MDR1 mRNA and Pgp (P <0.001) was significantly increased after transfection, but the expression of MDR1 mRNA and Pgp were significantly decreased (P<0.05). The expression in VCR group was significantly lower than that in siRNA-transfected cells+VCR group (P<0.01).C onclusion A20 siRNA could effectively enhance NF-kappa B expression in OCI-LY1 cells. NF-kappa B may up regulate the expression of its downstream genes such as MDR1 and cause apoptosis, in turn enhancing the inhibition of cell proliferation. VCR can reduce MDR1 mRNA and Pgp expression in OCI-LY1cells and the effect of VCR could be attenuated by A20 siRNA.
    Curcumin inhibits the apoptosis of HUVECs induced by oxidized low density lipoprotein
    2017, 37(5):  636-642. 
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    Objective To investigate the protective effect of curcumin on apoptosis of human umbilical vein endothelial cells (HUVECs) induced by oxidized low density lipoprotein (ox-LDL) and its possiblemechanism.Methods Cultivated HUVECs were divided into six groups: control group, oxLDLgroup,oxLDL plus endoplasmic reticulum stress(ERS) inhibitor PBA group,curcumin group, oxLDLpluscurcumingroup,oxLDL plus curcumin plus PI3K inhibitor LY294002 group.Cell viabilities were evaluated by CCK-8 assays .The proportions of apoptotic cells were assessed by flow cytometry .The translocation of activating transcription factor 6 (ATF6) abserved by laser confocal microscopy.Western blot was used to determine the expression of the ERS associated proteins:glucose-regulated protein 78( GRP78)、protein kinase-like ER kinase(PERK) 、inositol-requiring kinase1(IRE-1) and the related pathways protein:LOX-1、AKT and phophorylated AKT. Results Compared with control group,increased the proportions of apoptotic cells(P<0.01),enhanced the expressions of ERS related proteins(P<0.01),promoted the transfer of ATF6 into the nucleus,as well as increased the expression of LOX-1(P<0.01)and decreased the expression of p-AKT(P<0.01) in the oxLDLgroup;Compared with oxLDLgroup,PBAinhibited oxLDL-induced HUVECs apoptosis(P<0.01),curcumininhibited oxLDL-induced the expression of ERS associated protein and LOX-1(P<0.01), the nuclear translocation of ATF6, the apoptosis of HUVECs (P<0.01), and it also increased oxLDL-induced down-regulation of p-AKT expression (P<0.01); LY294002partially attenuated the inhibitory effect of curcumin on ERstress-related protein expression induced by oxLDL(P<0.05). Conclusions Curcumin can reduce oxLDL induced apoptosis of HUVECs, its mechanism may be through the inhibition of LOX-1 expression and activation of AKT pathway to reduce ERS in cell.
    Netropsin inhibits invasion and migration ability of gastric cancer cells
    2017, 37(5):  643-647. 
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    Objective To investigate theeffect of netropsin on migration and invasion ability of gastric cancer cells and its mechanisms. Methods To determine if netropsin inhibits migration and invasion of gastric cancer cells, Transwell migration and invasion assay was performed. Then Western blot was performed to detect expression of E-cadherin and vimentin in gastric cancer cells with or without presence in medium netropsin. Finally,immunofluorescence was performed to detect changes in the cellular localization of β-catenin to validate whether Wnt/β-catenin pathway was suppressed or not.Results Netropsinwith the concentration of 25 μmol/L had minimal inhibition effect on cell proliferation and was able to suppress ability of migration and invasion by inhibitingEMT in gastric cancer cells(P<0.05).Meanwhile netropsin was able to down-regulated the expression of epithelial markers E-cadherin and up-regulated the expression of mesenchymal marker vimentin. Finally, immunofluorescence showed that netropsin was able to block translocation of β-catenin from cytoplasm to nuclear.Conclusions Netropsin can inhibit EMT thereby suppressing migration and invasion of gastric cancer cells.The mechanism is that netropsin can compete with HMGA2 for transcription factor binding site thereby suppressing the Wnt/β-catenin pathway.
    Isolation and amplification of hepatic progenitor cells from fetal mouse in vitro
    2017, 37(5):  648-652. 
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    Objective To isolate and cultivate mouse hepatic progenitor cells (mHPCs) from E14.5 mouse fetal liver in vitro and induce mHPCs differentiation into cholangiocytes. Methods Isolation of mHPCs from mouse fetal liver wasbased on the cell surface antigendelta-like protein 1/preadipocyte factor 1 (Dlk/Pref-1) by a fluorescence-activated cell sorter (FACS). Then mHPCsisolated wereco-cultured with/without mouse embryonic fibroblasts (MEFs) by using Transwell. The cellantigen alpha-fetoprotein (AFP), albumin (ALB) and cytokeratin19 (CK19) expression in fresh isolatedDLK1+cells or co-cultured for 4 days and 6 days were observed with immunocytochemical method. Results When co-cultured with MEFs, the division and proliferation were observed in most of DLK1+ cells andgrape-like aggregation was formed. Cells began to adhere to growth and began to become spindle-shaped on 4th day.TheDLK1+cells isolated freshly by FACS were expressed AFP and low levels of ALBbut not expressed CK19. But, these cells expressed CK 19and weak expression of ALB on4thday.In addition, the expression of CK19increased and the expression of ALB almost not detected on 6th day. Conclusion Most of Dlk1+ cells, isolated from E14.5 fetal livers by FACS, wereprovedto be mHPCs. Furthermore, these cells can proliferate quickly and differentiate into cholangiocytes by co-culture with MEFs.
    Expression of amyloid beta peptide and its related metabolic enzymes in the brain of mice with Alzheimer’s disease and type 2 diabetes mellitus
    2017, 37(5):  653-657. 
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    Objective To examine the changes of Aβ expression and its related metabolic enzymes in the brains of AD and T2DM mice, so as to explore the possible mechanism of type 2 diabetes mellitus combined with AD. Methods Five-month-old APP/PS1 double transgenic mice, ob/ob mice and the wild-type control mice were employed in this study. Immunohistochemical (IHC) staining, Elisa and Western blot were used to detect SP, Aβ and its related metabolic enzymes. Results A certain number of SPs were observed in the cerebral cortex and hippocampus of APP/PS1 mice; SPs were occasionally observed in the cortex of ob/ob mice, while no SP appeared in wild-type mice. Aβ40 and Aβ42 levels were significantly increased in APP/PS1 and ob/ob mice brains as compared with controls (P<0.05), thought both Aβ40 and Aβ42 levels in AD mice were significantly higher than those of ob/ob mice (P<0.05). APP expression level was highest in APP/PS1 mice among 3 groups, and its expressed higher in ob/ob mice than that of control mice (P<0.05). BACE1 expression was notably increased in APP/PS1 and ob/ob mice as compared with control(P<0.05), however, it expressed higher in APP/PS1 mice than ob/ob mice (P<0.05). The expression of Aβ degradation enzyme IDE was reduced in APP/PS1 and ob/ob mice(P<0.05), while lowest in ob/ob mice. Conclusions Abnormal production, degradation and gathering of Aβ are not only occurred in the early stage of AD, but also in the brain of T2DM. These results indicate that overexpression of Aβ may be one of main reasons for T2DM combined AD.
    Anticonvulsant effect of ceftriaxone in temporal lobe epilepsy
    jing li
    2017, 37(5):  658-662. 
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    Objective To study the effects of Ceftriaxone on the seizures and the expression of glutamate transporter (GLT-1) in kainic acid (KA) epilepsy model. Methods Firstly, a chronic spontaneous seizure mouse model was established by unilateral hippocampal injection of KA and monitored by vEEG technique to record seizures. The experimental group received intraperitoneal injection of Cef 200 mg/(kg?d) and the control group received normal saline. Several indicators such as seizure frequency, interictal spike waves and histological phenotypes were used to evaluate the function of Cef. Then we use the western blot to detect the effect of expression for GLT-1. Results Unilateral hippocampal injection of KA 200ng successfully established the mesial temporal lobe epilepsy model. Cef can reduce the seizures from 2.145 times / day to 1.597 times / day, decreased by 31.2% with a statistical significance (p<0.05). Cef treatment did not significantly enhance the expression of GLT-1which test by Western blot. Conclusion Intraperitoneal injection of Cef partially inhibit the seizures of KA model, but the expression of GLT-1 in hippocampus was not enhanced. It is suggest that ceftriaxone may inhibit seizures through other mechanisms.
    Effect of p53 with R213 mutantion on the expression of p21 in colorectal cancer cells HCT116
    2017, 37(5):  663-667. 
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    Objective To determine p53 with R213 mutation impacts on the expression of its downstream target gene p21 in HCT116-/- colorectal cancer cell line. Methods Mutations of p53 at R213Q (R/Q) and R213K (R/K) were respectively constructed and transfected into the p53 null HCT116 cells. The promoter activitydrivedby p53 was detected by dual-luciferase reporter system. EMSA assay was used to detect the ability of p53 binding to the promoter of its target gene p21.The expression of p53 downstream target genes p21 wasexamined by Real time PCR and Western blot experiments. Results The mutation of R213K disrupted the efficiency of p53 transactivation(P<0.05). The mutants were less efficient in targeting the consensus binding sequences of p53 in the regulatory region of p21 gene(P<0.05), In addition, The R/K mutant attenuated the expression of p21gene(P<0.05). The mobility of R213Q was increased.Conclusion The different mutations of Arginine 213 have different impact on p21 in colorectal cancer cells. It could exist diverse mechanism to regulate.
    IL-1β promotes glial scar formation after spinal cord injury in rats via JAK2-STAT3
    2017, 37(5):  668-675. 
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    Objective To investigate the mechanism of IL-1β in promotingglial scar formation after spinal cord injury. Methods The experimental model of SCI was created by extradural compression of the spinal cord using an aneurysm clip. Rats were randomly divided into model group, sham operation group, IL-1β inhibitor IL-1RA group, IL-1β group and IL-1β+JAK2-STAT3 inhibitor AG490 group, according to different interventions, then were given normal saline, IL-1RA, IL-1β and IL-1β+AG490 every 10μL respectively, sham group received only laminectomy. The motion function of the hindlimbs of rats was measured by Basso Beattie Bresnahan(BBB) scores and the expression of GFAP , vimentin and P-STAT3 were detected by Western blot technique, immunofluorescence assay and immunohistochemistry technique at corresponding time points(at the 8th , 12th hour, 1st, 3rd, 7th and 14th day after SCI). Results The expression trend of P-STAT3(at the 8th and 12th hour after SCI), GFAP and vimentin(at the 7th and 14th day afterSCI)was: the expressions of P-STAT3, GFAP and vimentin in the model group were significantly higher compared with the sham group(p<0.01), the expressions of P-STAT3, GFAP and vimentin in the IL-1RA group were significantly lower compared with the model group(p<0.05) whereas significantly higher compared with the sham group(p<0.05); the expressions of P-STAT3, GFAP and vimentin in the IL-1β+AG490 group were significantly lower compared with the model group(p<0.05)whereas significantly higher compared with the sham group(p<0.05), the expressions of P-STAT3, GFAP and vimentin in the IL-1β group weresignificantly higher compared with the model group(p<0.05). Conclusions IL-1β can improve glial scar formation via JAK2-STAT3 signal, inhibition of IL-1β or JAK2-STAT3 can reduce glial scar formation and promote functional recovery of spinal nerve.
    rs4715986 in DTNBP1 is associated with paranoid schizophrenia in Chinese Han population
    2017, 37(5):  676-681. 
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    Objective The present study aimed to investigate association of the DTNBP1 gene polymorphisms with paranoid schizophrenia in Han Chinese populations by a two-stage study. Methods In the discovery stage, 11 SNPs in DTNBP1 were tested in 532 patients with paranoid schizophrenia and 488 controls by Sanger sequencing. In the validation sample (1 111 cases and 1 435 controls), Taqman genotyping method was used to genotype SNP rs4715986. Results In the discovery stage, SNP rs4715986 showed disease association (χ2 = 11.02, P < 0.01) with paranoid schizophrenia for analysis of 532 patients and 488 controls; such an association survived a correction with 10,000 permutations (corrected P < 0.05). In the validation sample (1 111 cases and 1 435 controls), we confirmed the genotypic association (χ2 = 9.292, P = 0.01) of rs4715986 with paranoid schizophrenia. Conclusions The present results suggest that the DTNBP1 gene may be mainly involved in the development of paranoid schizophrenia in the Chinese Han population.
    Association between the SNPs in microRNA binding sites of ABCG5/8 and glucose and lipid metabolism during pregnancy
    2017, 37(5):  682-686. 
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    Objective The aim of the study is to explore the relationship between the SNPs in microRNA binding sites of ABCG5/8 and the glucose and lipid level during pregnancy. Methods 1925 pregnant women were recruited at Peking Union Medical College hospital from 2006 to 2011. The clinical data were collected and the total genomic DNA was extracted from whole blood samples. ABCG5/8, which was reported to be related with the glucose and lipid metabolism closely, were selected as the candidate gene and the SNPs in its microRNA binding sites with minor allele frequency > 5% in Han Chinese in Beijing were chosen. Then the genotyping was performed and analyzed. Results There was only one SNP matching the criteria, rs2278356, and it is significantly associated with LDL-C and TC level during pregnancy (LDL-C: b = 0.104 mmol/L, 95%CI 0.023-0.185 mmol/L, p<0.05;TC: b = 0.105 mmol/L, 95%CI 0.080-0.203 mmol/L, p<0.05). Conclusions We found the association between rs2278356 in 3’ UTR of ABCG5/8 and the LDL-C and TC level in pregnant Chinese Han women, which may provide the individualized treatment strategy for pregnant women with high cholesterol.
    Monocrotaline increases expression of IL-1β and IL-18in lung tissue of pulmonary hypertension rats
    2017, 37(5):  687-690. 
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    Objective To explore the change of IL-1β and IL-18 expression in pulmonary hypertension induced by monocrotaline. Methods Divide the mouses into two groups, normal control group and experimental group (n=10). Establish ratspulmonary hypertension model induced by monocrotaline. Detect the model by ultrasound, myocardial cells HE dyeing and tunnel test; ELISA was used to detect the serum biological markers’ level of NF-κB, COX2, IL-6, IL-1β, TNF-αand NO; Immunohistochemical was used to detect the expression level of IL-1β and IL-18 in the lung tissue; the protein change of NLRP3 in the lung tissue was detected by Western blot. Results Serum biological markers’ level of NF-κB, COX2, IL-6, IL-1β, TNF-α and NO are significantly increased in PAH rats (P<0.05);The expression ofIL-1β, IL-18 in the lung tissueincreased obviously (P<0.05); The NLRP3 protein expression was significantly higher in experimental group. Conclusions Changes of NLRP3 effect the increasing expression of IL-1β and IL-18 plays an important role in pulmonary hypertension induced by monocrotaline.
    Lunasin alleviates the sports articular cartilage injury of rats
    2017, 37(5):  691-695. 
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    Objective To exploer the function and underlying mechanism of Lunasin on sports articular cartilage injury. Methods Wistar rats were randomly divided into negative control group and model group; after injection molding for 3 times, the model rats were feed for 28 days. Then the model rats were divided into sham model group, 0.01, 0.05 and 0.1 mmol/L Lunasin treatment group respectively. After treatment, ELISA was used to analyze the production of IL-1β、IL-6、TNF-α、MMP-6 and MMP-8. The SOD activity and iNOS were evaluated by their ELISA kit. Western blot was used to detect the expression of NRF2, Keap1, LC-3Ⅱ, Bax, Beclin1, p-AMPK, AMPK. Results Compared with control, the dose of IL-1β、IL-6、TNF-α、MMP-6 and MMP-8 in serum of model rats were significantly increased (P < 0.05), however, after treatment with lunasin for 1 month, these inflammatory factors were obviously reduced then that of model rats (P < 0.05); Furthermore Lunasin treatment obviously increase SOD activity、up-regulate NRF2 expression and down-regulate the generation of nitric oxide synthase (iNOS) (P < 0.05); Additionally, Lunasin can raise the expression of autophagy-related protein(beclin1 and LC-3Ⅱ), reduce the expression of apoptosis protein (Bax) in damaged articular cartilage. Conclusion Lunasin benefit the repair of damaged joints by reducing the production of inflammatory mediators, activating of oxidative stress system and autophagy pathway.
    TRPC1/ORAI1 requlates Ca2+ entry and NO generation mediated by HUVEC SOC and ROC
    2017, 37(5):  696-704. 
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    Objective To study the function of TRPC1/ORAI1 in store and receptor-operated Ca2+ entry and nitric oxide generation by SOC and ROC in human umbilical vein endothelial cells. Methods HUVECs were collected and cultured to the second~third passage. We silenced the expression of their genes in HUVEC by transfection constructed TRPC1 or ORAI1 RNA interference plasmids. The interference efficiency of their protein and mRNA levels were determined by Western blotting and real-time PCR, respectively. The cells were divided into: short hairpin RNA(shRNA) group; control group and vehicle group. The cells were incubated with CaR agonist, CaR negative allosteric modulator Calhex231 and receptor-operated channels (ROC) analogue TPA, Protein kinase C (PKC) inhibitor Ro31-8220, PKCs and PKCμ inhibitor Go6967 incubate, intracellular Ca2+ concentration ([Ca2+]i) was detected using the fluorescence Ca2+ indicator Fura-2/AM, the production of NO was determined by DAF-FM of every group in HUVEC. HUVECs were transduced with shRNA-TRPC1 and shRNA-ORAI1 at same time, after cultured with CaR agonist, [Ca2+]i and the production of NO was determined. The interaction between ORAI1 and TRPC1 were examined by Co-immunoprecipitation. Results 1) Compared with control group, shRNA targeted to the TRPC1 or ORAI1 genes decreased their mRNA and protein levels, respectively (p<0.05); 2) In four different treatment under the action of factors, the [Ca2+]i and the net NO fluorescence intensity ratio values of transfection of TRPC1 or ORAI1 shRNA group were significantly reduced (p<0.05). 3) Compared with the control and single shRNA group, the[Ca2+]i and the net NO fluorescence intensity ratio values of transfection of TRPC1 and ORAI1 shRNA group were significantly reduced (p<0.05). 4) ORAI1 co-precipitates with TRPC1, indicating internation of a molecular complex had enhanced by CaR agonist. Conclusions TRPC1, ORAI1 are components of SOCE and ROCE channels in store and receptor-operated Ca2+ entry and nitric oxide generation in human umbilical vein endothelial cells.
    Relationship between the expression of CEACAM1, CD34 and the metastasis of gastric cancer
    2017, 37(5):  705-708. 
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    Objective To investigate the prognostic impact of carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) and CD34 on invasion and metastasis of gastric cancer. Methods CEACAM1 and CD34 expressions were detected by immunhistochemistry in 90 cases of gastric cancer and 30 cases of normal gastric mucosa. Then the relationship among CEACAM1, MVD marked by CD34 antibody and gastric cancer patients’ clinical pathological features and prognosis were analyzed. Results The positive rates of CEACAM1 and CD34 protein in gastric cancer were significantly higher than those in normal gastric mucosa (P<0.05). The positive degree of CEACAM1 was closely associated with histological differentiation, tumor invasion, nodal metastasis and TNM stage. MVD marked by CD34 was related with tumor size, histological differentiation, tumor invasion, nodal metastasis and TNM stage(P<0.05). Conclusions In gastric cancer, CEACAM1 and CD34 promote its capability of invasion and metastasis, and they might be used as an indicator in predicting development and prognosis.
    Relationship between polymorphism of FcγRIIIA F158V genotype and systemic lupus erythermatosus as well as lupus nephritis in Chinese Han population
    2017, 37(5):  709-713. 
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    Objective To study the frequencies of FcγRIIIA polymorphisms in Chinese people and the potential role of these polymorphisms(SNPs) in systemic lupus erythermatosus(SLE) and lupus nephritis(LN). Methods We studied 324 unrelated Chinese SLE patients and 319 controls. FcγRIIIA F158V polymorphism was genotyped by specific primer-polymerase chain reaction(PCR). Results The observed frequency of F allele of FcγRIIIA polymorphism is 60.8% in Chinese people. FcγRIIIA polymorphism revealed a significant difference of both genotype and allele distribution in SLE patients and controls. LN patients have a significant increase in carrier frequency of the FF genotype, while the VV genotype reveals a significant decreased risk of early onset of LN. Conclusion FcγRIIIA polymorphisms might be related with SLE in Chinese people. The homozygous FF genotype of FcγRIIIA might be the risk factor for LN, while the homozygous VV genotype might be the protective factor for late onset lupus nephritis.
    Increased gastroesophageal junction compliance in patients with non-erosive reflux disease
    2017, 37(5):  714-717. 
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    Objective To explore the biomechanical mechanism of non-erosive reflux disease(NERD). Methods From October 4,2014 to September 30,2015,17 NERD patients and 17 healthy controls were recruited from the outpatients department of China-Japan Friendship Hospital.The cross-sectional area(CSA)、intra-balloon pressure (IBP)and compliance((Δv/Δp)of each were measured by endoscopy in combination with EndoFlip. Results With the increase of balloon volume,the compliance of gastroesophageal junction in NERD group was significantly higher than that of control group(p<0.01) while the cross sectional area and intra-balloon pressure were similar.Conclusion Increased compliance of the gastroesophageal junction might be a factor in the pathogenesis of non-erosive reflux disease.
    Effects of forced-air prewarming combined with fluid warming on body temperature and thermal comfort in patients undergoing lobectomy
    2017, 37(5):  718-722. 
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    Objective To investigate the effects of forced-air prewarming combined with fluid warming on body temperature and thermal comfort in patients undergoing lobectomy. Methods forty six patients scheduled for Video-assisted Thoracoscopic Surgery (VATS) of lobectomy were randomized into two groups (n=23 each):warming group (group T) and the control group (group C). Forced-air prewarming combined with infusion heating was applied in the warming group,while only conventional passive warming was used in control group. The tympanic membrane temperatures were recorded before prewarming,after prewarming, during the anesthesia, the end of operation, the moment in and out of the Postanesthesia care unit (PACU). Incidence of postoperative shivering, thermal comfort and the time in the PACU were recorded. Results Compared with control group,The warming group had a lower temperature decrease at 1 hour after operation, 2 hours after operation, 3 hours after operation and end of operation( P< 0.01), warming group had significantly lower incidence of hypothermia and shivering than control group(8.7% vs 56.5%,4.3% vs 34.8%,P<0.05),and the thermal comfort score was higher in warming group than in control group(8.52±0.94 vs7.65±0.83,P<0.05). Conclusions Forced-air prewarming combined with fluid warming has significant clinical effects in helping to stabilize patients’ body temperature during operations, reduce the incidence of hypothermia and shivering and improve the thermal comfort, which provides a simple and effective temperature protection strategy for patients undergoing lobectomy.
    Hemispherotomy for hemispheric epilepsy: outcome and early follow up for complications
    2017, 37(5):  723-725. 
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    Objective To investigate the results and complications of hemispherotomy for drug resistant hemispheric epilepsy. Methods The authors reviewed 5 patients who were diagnosed as drug resistant hemispheric epilepsy and operated in the neurosurgery department of Peking Union Medical College Hospital from 2011 to 2013. All the 5 patients were underwent hemispherotomy after carefully multidisciplinary pre-operation evaluation. Results All patients tolerated the procedure well and the postoperative course was smooth. All the 5 patients were seizure free in the period of following up of 46 to 69 months. Conclusions According to literatures and the authors’ experience, hemispherotomy is as safe and efficient for hemispheric epilepsy as hemispherectomy. It is the most important modification for hemispherectomy. The procedure of hemispherotomy is complex but not very difficult, Illustrating a good prospect of application and extension.
    Gut microbiota and cardiovascular disease
    2017, 37(5):  729-733. 
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    Microbiota within the intestines and the host interact with each other. Resently,the association and causality between gut microbiota and cardiovascular diseases have been well studied. Some underlying mechanisms,especially metabolic pathways are known, including the role of trimethylamineN-oxide in the pathogenesis of atherosclerotic heart disease and heart failure and that of short chain fatty acids and their receptors in blood pressure regulation.
    Integration of judicial expertise into basic medical education improves the growing up of excellent medical doctors
    2017, 37(5):  734-737. 
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    Through the addition of discussion course associated with judicial expertise during the pre medical education, integration of true and typical forensic pathological cases into basic medical theory and experimental education, further addition of optional course of ,and guiding the medical students applying the scientifically training projects about forensic pathology, students may improve their learning interesting and clinical thought, and are made early warning and increase the abilities of preventing and dealing with the suddenly medical tangles in the future, at the same time, the medical teachers also increase their professional levels and teaching qualities. These benefit the growing up of high quality medical doctors, decrease and even prevent the happening of medical tangles.
    Improvement of comprehensive abilities of eight-year program undergraduates by complex scene PBL teaching method
    2017, 37(5):  738-741. 
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    Objective Evaluation the effect to improve the eight-year program medical students' comprehensive ability between standard of PBL teaching mode and complex scenarios PBL teaching mode .Methods To comparative improved medical teaching effect between complex scenarios PBL teaching mode and standard of the PBL teaching mode by using the final exam scores and DREEM scale. Results The complex scenarios PBL teaching mode improve the ability of problem solving score significantly higher than the standard scores of PBL teaching mode at the start of analyzing questions and help students to stimulate interest, increase self-confidence, (p < 0.05). Comparative analysis shows the comprehensive thinking ability of medical students has improved significantly. Conclusions Complex scenarios PBL teaching mode can more improve the ability of handling the students' study enthusiasm, comprehensive ability and strain capacity than that of standard PBL teaching mode. The obvious effect is worth promoting.
    Encoding of experimental instruments and equipment fixed assets property in basic medical sciences research
    2017, 37(5):  742-746. 
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    In order to standardize the management of fixed assets in basic medical research, and to solve the problem of " one equipment with more than one code ", we discussed the fixed assets coding of instrument and equipment in this paper. The existing equipment classification of the Institute of Basic Medical Sciences of the Chinese Academy of Medical Sciences was analyzed. Depending on application of the experiment and the principle of equipment, the novel fixed assets encoding dictionary of instrument and equipment was generated, which could fit in with the actual application and development of basic medical research. The managers could find the corresponding code quickly with the standardized equipment name. The new encoding dictionary could ameliorate the classification of basic medical experimental instruments, avoid multiple coding situations of equipment and improve the management work in the future.