Table of Content

05 June 2017, Volume 37 Issue 6

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ADAR2 inhibits MAVS expression by editing its 3’-UTR in cells

2017, 37(6):  747-751. 

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Objective To investigate the effect of ADAR2 on the expression of MAVS and its mechanism. Methods The RT-qRCR was used to detect the expression of ADAR gene family, the expression level of ADAR2 and MAVS in cells. The effect of ADAR2 on the 3 'UTR region of MAVS was detected by Pyrosequencing. To detect the expression of luciferase by dual luciferase reporter plasmid assay; The expression of ADAR2 and MAVS were detected by Western blot. Results In the ADAR family, the abundance of ADAR1 was the highest, followed by ADAR2, but the expression of ADAR3 was poor, which was almost impossible to detect（P < 0.05）. ADAR2 plays a critical role in RNA editing of chr20:3869744 sites on the 3 'UTR region of MAVS（P < 0.001）. On the 3 'UTR editing site of MAVS, the luciferase activity of the edited G allele was significantly lower than that of the normal A allele（P < 0.01）. At the level of transcription and translation, ADAR2 significantly inhibited the expression of MAVS（P < 0.05）. Conclusions ADAR2 by editing MAVS' 3 'UTR on the chr20:3869744 site, which makes the occurrence of A to G replacement, inhibits the expression of MAVS.

Optimization of in vitro amplification of peripheral blood γδ T cells with cytokines

2017, 37(6):  752-757. 

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Objective To optimize in vitro amplification of human γδ T cells with cytokines for tumor adoptive immunotherapy. Methods On the basis of the immobilized anti-TCR γδ antibody plus IL-2 system, other γ chain receptor family cytokines, including IL-7, IL-15 and IL-21, were tested to amplify human peripheral blood γδ T cells either alone or in diversity combination. The percentage of γδ T cells was measured by flow cytometry, and the proliferation efficiency of γδ T cells was calculated. The expression of proliferation- or cytotoxicity-related molecules on γδ T cells was examined by flow cytometry in order to explore the relevant mechanisms. The cytotoxicity of γδ T cells to Daudi cells was detected by lactate dehydrogenase. Results IL-15 alone but not IL-7 or IL-21 could increase the γδ T cell purity, amplification efficiency and cytotoxicity to reach comparable levels to those of IL-2. IL-2 plus IL-15 up-regulated the expression of CD69 on γδ T cells and significantly increased their amplification efficiency (P<0.05). IL-2 plus IL-21 enhanced the cytotoxicity of γδ T cells against Daudi cells by increasing the expression of granzyme A (P<0.001). The combination of IL-2, IL-15 and IL-21 could significantly improve the cytotoxicity of γδ T cells but reduce their amplification efficiency. In addition, when IL-21 was applied for a short time, it could also enhance the cytotoxicity of γδ T cells (P<0.05). Conclusion The combination of IL-2 and IL-15 as well as a short time addition of IL-21 is the best cytokine recipe to amplify human peripheral blood γδ T cells in vitro with immobilized anti-TCR γδ antibody, which can increase both the proliferation efficiency and the cytotoxicity to tumor cells of γδ T cells.

Vinblastine enhances the expression of tumor resistance gene abcb4 in zebrafish

2017, 37(6):  758-762. 

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Objective To explore the effects of vinblastine on abcb4 tumor resistance gene expression in early development zebrafish and lay an important foundation for multi-drug resistant antineoplastic screening in zebrafish model. Methods Zebrafish embryos of 0.5~1.5 hours after fertilization(0.5~1.5 hpf) were exposured by different concentrations of vinblastine, and then calculated the IC50 of vinblastine. Zebrafish embryos of 0.5~1.5 hpf were treated with different concentrations of vinblastine and embryo culture medium respectively, and then observed zebrafish embryo development in 24~120 hpf and recorded the number of death, hatch and malformation .Evaluating the impact of vinblastine on abcb4 gene expression in zebrafish with quantitative real-time PCR and whole-mount in situ hybridization by collecting zebrafish embryos exposed to different concentrations of vinblastine. Results Vinblastine IC50 in zebrafish embryos is 3.08 ?mol/L. The mRNA level of abcb4 gene in vinblastine treated embryos was significantly increased compared to blank control group. Moreover, abcb4 gene positive hybridization signals were found in the small intestine of zebrafish embryos after 120 hpf and also partly found in the brain and heart of zebrafish embryos by the method of whole-mount in situ hybridization. Conclusions Vinblastine can significantly increase the expression level of abcb4 tumor resistancegene in early development zebrafish embryos, which points out that zebrafish can be used as a tumor resistant drug screening model.

High pressure level in human airway enhances the expression of 3 cytokines related to airway remodeling

2017, 37(6):  763-767. 

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Objective Under the circumstance of mechanical ventilation, we designed to explore the relationship between different airway pressure levels and expression of related cytokines to airway remodeling. Methods Fourty-two chronic obstructive pulmonary disease cases (COPD group) and thirty-three without chronic airway inflammatory dieases (control group) were collected. The above cases underwent mechanical ventilation in the period of general anesthesia. According to different levels of peak inspiratory pressur e (PIP) , above two groups, randomly and respectively, were divided into high PIP (HPIP, 24 cmH2O) group, moderate PIP (MPIP, 22 cmH2O ) group, low PIP (LPIP, 20 cmH2O ) group. All positive end expiratory pressure (PEEP) was set to 5 cmH2O. Then the collection of BALF was performed before and 3 hours after applying ventilator. The related factors to airway remodeling, such as fibroblast growth factor 2 (FGF-2) , transforming growth factor β1 (TGF-β1) and matrix metalloproteinase-9 (MMP-9) , were measured by enzyme-linked immunosorbent assay (ELISA) and Western blot. Results 1) Before mechanical ventilation, the levels of FGF-2, TGF-β1 and MMP-9 in COPD group were higher than control group (P<0.01) . 2) 3 hours after mechanical ventilation, we saw significant upregulated expression of FGF-2, TGF-β1, MMP-9 in HPIP subgroup in control group (P<0.05) and the 3 factors levels of COPD group were all increased (P<0.05) . Moreover, HPIP subgroup were significantly higher than MPIP and LPIP subgroup in COPD group (P<0.05) . 3) The expression of FGF-2, TGF-β1, MMP-9 in BALF had a positive correlation with the airway pressure levels in COPD group(P<0.01). Conclusions Under mechanical ventilation, sustained high airway pressure may enhance the expression of FGF-2, TGF-β1 and MMP-9 which could result in airway remodeling by mechanosensitive cation channel in bronchial epithelial cells, especially in COPD patients.

Effects of combination of DLX1 and BMP9 on inducing osteogeninc differentiation of human osteosarcoma cells

2017, 37(6):  768-774. 

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Objective To investigate the effects of Distal-less homeobox 1 (DLX1) and bone morphogenetic protein 9 (BMP9) on inducing osteogeninc differentiation of human osteosarcoma cells. Mtethods The MG63 cells were infected with AdDLX1 and AdBMP9 alone or in combination, grouped as DLX1 group(AdDLX1+AdRFP)、DLX1+BMP9 group(AdDLX1+AdBMP9)、BMP9 group(AdBMP9+AdRFP)、RFP group(AdRFP). The expression of DLX1 and BMP9 were detected by RT-PCR and Western blot. The cell migration and invasion were analyzed by Transwell assay. Alkaline phosphatase(ALP) staining and numeration were used to detect the early osteogenic potential of MG63 cells, and the late osteogenic potential was detected by the alizarin red staining. The expression of osteopontin (OPN) protein was determined by Werstern blot assay. ResuLts The mRNA and protein expression of DLX1 and BMP9 were increased by AdDLX1 and AdBMP9（P＜0.05）. Transwell assay resμLts showed the cell migration and invasion of DLX1 group、DLX1+BMP9 group and BMP9 group were decreased, DLX1+BMP9 group was the most one (P < 0.05). The activity of ALP and calcium deposition as well as OPN protein were increased in DLX1+BMP9 group and BMP9 group, what’s more, these markers in DLX1+BMP9 group were more obvious (P < 0.05). Conclusions The combination of DLX1 and BMP9 could inhibit the malignant biological behavior and induce osteogenic differentiation of osteosarcoma cells.

Over expression of TMEM66 attenuates the neointima formation of carotid artery after balloon injury in rats

2017, 37(6):  775-780. 

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Objective To clarify the role of transmembranprotein 66(TMEM66) in neointima formation of carotid artery induced by balloon injury in rats. Methods SD rats were randomly divided into three groups (n=10), including control group, the left carotid artery balloon injury group and injury group added with lentiviruses, respectively. Accordingly, the neointima formations of carotid arterywere detected by H/E staining;the expressions of TMEM66 in carotid arteryinjuried by balloon.were determined by western blot,q-PCRand immunohistochemistry, and the migration and proliferation of VSMCs were measured by scratch test and CCK8, respectively. Results Compared with control group, the expression of TMEM66 in carotid artery were obviously decreased after balloon injury (P＜0.05). Most importantly, the neointima formation of carotid artery was significantly reversed by overexpression of TMEM66 via specific lentiviruses (P＜0.05). Meanwhile, it is obviously that upregulation of TMEM66 can alleviate the migration and proliferation of VSMCs by PDGF(P＜0.05). Conclusions TMEM66 is able to attenuate the neointima formation of carotid artery after balloon injury.

Effect of lncRNA-1700020I14Rik on the fibrosis in mouse mesangial cells under high glucose condition

2017, 37(6):  781-785. 

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Objective To construct the lncRNA-1700020I14Rikplasmid and detect its effect on the fibrosis of mice mesangial cell (MMC) cultured with high glucose medium. Methods RT-qPCR was used to measure the expression of 1700020I14Rikin MMC cultured with low glucose medium or high glucose. Total RNA was extracted from MMC and cDNA was got by RT-PCR. The whole fragment of lncRNA-1700020I14Rik amplified by PCR was constructed into plasmid pcDNA3.1(+) through PCR. Lipidosome 3000 was used to transfect the plasmid into the MMC cultured with high glucose medium and RT-qPCR was used to measure the expression level of 1700020I14Rik. Western blot was used to analysis the expressions of fibronectin, collagen Ⅳ and TGF-β1. Results 1700020I14Rik was significantly down-regulated in MMC cultured with high glucose and it was significantly up-expressed in the MMC after transfecting with pcDNA3.1(+)-1700020I14Rik. The expressions of fibronectin, collagen Ⅳ and TGF-β1 were down-regulated by 1700020I14Rik. Conclusion The plasmid pcDNA3.1(+)-1700020I14Rikis able to effectively express the lncRNA-1700020I14Rik. Over-expression of 1700020I14Rik may protect mesangial cells from fibrosis conduced under high glucose condition.

Down-regulation of osteocytic TGF-β/Smad4 inhibits the osteoblastic and osteoclastic differentiation in mouse BMSCs

2017, 37(6):  786-791. 

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Objective To determine the effect of ostecytic TGF-β/Smad4 signaling on osteoblastic and osteoclastic differentiation in bone marrow stromal cells (BMSCs). Methods Mice with osteocytic TGF-β/Smad4 conditional knock down (Smad4ot CKD) were generated as previously by crossing DMP1-8kb-Cre mice with Smad4lox(ex8)/lox(ex8) mice. The osteocytes were isolated from tibial and femoral diaphysis and co-cultured with wild-type BMSCs. ALP staining, Alizarin red staining and TRAP staining were performed to show osteoblastic and osteoclastic differentiation. Then, their marker genes were detected by qPCR and proteins measured by Western blot. Results The expression of Runx2 and Osterix were reduced in smad4 CKDot co-cultured with BMSCs compared with controls (P<0.01). Similarly, the specific markers of osteoblastic differentiation were decreased (P<0.01). Additionally, the expression of RANKL was not obviously changed in with BMSCs. However, OPG was highly expressed in control group compared with smad4 CKD ot co-cultured group (P<0.05). Thus, the radio of RANKL/OPG was significantly reduced (P<0.05). Furthermore, the expression of RANK was inhibited. Conclusions The terminally-differentiated osteocytes are the cells regulating bone metabolism, while down-regulation of osteocytic-TGF-β/Smad4 inhibits BMSC osteoblastic and osteoclastic differentiation.

Inhibitory effect of 5-FU and cisplatin mediated by miR-449b on the migration of HCC cells

2017, 37(6):  792-796. 

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Objective To investigate the impact of5-Fluorouracil (5-FU) and cisplatin on micrRNA-449b (miR-449b) expression in human hepatocellular carcinoma (HCC) and elucidate the molecular mechanism of 5-FU and cisplatin inhibiting the migration of HCC cells. Methods Real-time qPCR analysis was conducted to determine the expression of miR-449b in 50 HCC tissues. Real-time PCR assay was performed to detect the expression of miR-449b in HCC cells with 5-FU and cisplatin treatment. The migration of HCC cells with the overexpression of miR-449b was determined by wound-healing assay; Rescue assay was employed to investigate the correlation between 5-FU&cisplatin, miR-449b and the migration capacity of HCC cells; The putative targets of miR-449b were predicted and validated using target prediction programs and immunoblots. Results The expression of miR-449b decreased in HCC tissues (P<0.0001). MiR-449b expression increased in HCC cells upon the treatment of 5-FU and cisplatin (P<0.001). The overexpression of miR-449b inhibited the migration of HCC cells (P<0.001). Rescue assay revealed that inhibition of miR-449b to prevent 5-FU and cisplatin induction resulted in suppressed migration in SMMC7721 cells (P<0.05). Catenin-δwasa functional target of miR-449b. Conclusions 5-FU and cisplatin inhibit the migration of HCC cells at least partly via inducing the expression of miR-449b.

Analysis of gene mutations and clinical phenotype in 3 patients and their families with osteogenesis imperfecta

2017, 37(6):  797-801. 

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Objective To improve the diagnosis and recognition of osteogenesis imperfecta（OI）, the genes of three families with OI were screened and its clinical phenotype and gene mutations were analyzed. Methods Clinical data were collected and then blood samples of patients and their families were extracted, high-throughput sequencing, generation sequencing, analysis of results. Results Proband1, FKBP10, Exon6,c.1016G＞A,R339Q,Homozygous mutation, heterozygous mutation of the same locus of his parents. Proband2, COL1A1,EXON9,c.671G>A,p.G224D,heterozygous mutation, heterozygous mutation of the locus of her father, non-mutation of the locus of her mother.Proband3,COL1A1,EXON30,c.2010delT,p.P670fs,heterozygous mutation, heterozygous mutation of the same locus of his mother, non-mutation of the locus of his father. Conclusions The novel mutation of FKBP10 may lead to the onset of XI type of osteogenesis imperfecta(OI) and the new phenotype. The relationship of two loci mutation of COL1A1 and OI was confirmed further.

Exosome extracted from hAMSCs promotes neurological function recovery after taumatic brain injury in rats

2017, 37(6):  802-807. 

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Objective: To investigate the therapeutic effect of exosomes extracted from human adipose-derived mesenchymal stem cells on traumatic brain injury (TBI) and its possible mechanism. Methods: Mesenchymal stem cells(MSCs) were isolated from healthy human adipose tissue and the exosomeswere extracted by ultrafiltration. Rats were divided into four groups: sham group, PBS control group, MSCs treatment group and exosomes treatment group. 24 h After TBI, the treatment group was injected locally along the lesion area, 30μL of PBS, 2 ⅹ105MSC, 25μg protein of exosomes respectively, the total volume was30μL. We performed the Modified Neurological Severity Score(mNSS) and the forelimb Foot-Fault Test in all rats beforeinjury and at 1, 3, 7, 10, 13, 16, 21, and 30 days after TBI. The rats were sacrificed at 3 and 7 days after TBI respectively，total RNA was extracted from rat brain tissue. The expression of TNF-α and IL-1β were detected by quantitative PCR. The rats were also killed at 30 days after TBI for testing the neuronal apoptosis in lesion area by tunel-neun double immunofluorescence. Results:exosomes treatment significantly promoted the recovery of neurological deficits caused by TBI，and the therapeutic effect was similar to MSCs， its possible mechanism may be the inhibition of the acute inflammation and the reducing of the neurons apoptosis after TBI.

Anti-HBs level within 15 years after vaccination in adult and newborns

2017, 37(6):  808-811. 

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Objective To investigate the anti-HBs level within 15 years after vaccination in adults and newborns in Beijing and provide the suggestion for the adult hepatitis B (HB) immunization plan. Methods A serological survey was conductedin 6705 subjects aged > 1 year old by multistage randomized cluster sampling in Beijing during August 2013 to February 2014. The subjects whohad received a 3-dose recombinationHB vaccine when they were newborns or adults aged ≥15 years old and did not undergo revaccination were selected. Antibody to hepatitis B surface antigen (anti-HBs)titers and positive rates within 15 years after vaccination wereevaluated. Results A total of 129 and 493 subjects who were vaccinated in adults and newborns were enrolled in the study. Based on the self-limited rate, 30%, of HBV infection among the general population aged 15 to 59 years, anti-HBs positive rates for the subjects vaccinated in adults were estimated to be 58.6%，62.5%and 48.4%during 0 -4, 5 - 9 and 10 - 15 years after vaccination respectively. The corresponding median of anti-HBs titers were 288.8, 120.6 and 62.6mIU/mL.The anti-HBspositive rates for the subjects vaccinated in newbornsduring 0 - 4, 5 - 9 and 10 - 15 years after vaccination were 83.3%, 47.3% and 43.5%, respectively. The corresponding anti-HBs titerswere 71.8, 8.9,and 6.7mIU/mL. Conclusions The protection afforded by primary immunization with recombination vaccine in adults and newborns lasts at least 15 years.

Association of the exposure of SO2 and NO2 with diabetes: a case-crossover study

2017, 37(6):  812-816. 

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Objective To assess the association of the concentration of SO2 and NO2 with diabetes in Nanjing. Methods We collected air pollution, meteorological data and diabetes-related deaths in Nanjing from 1 January 2008 to 31 December 2011 and conducted a time-stratified, bi-directional case-crossover design to examine theassociation. Subgroup analyses were performed after stratification by age, gender and season. Results The ORs of the diabetes-related mortality associated with each10μg/m3 elevation of SO2 and NO2were approximately 1.031 (95% CI: 1.005, 1.058), 1.042 (95% CI: 1.012, 1.074). SO2 and NO2 were significantly associated with diabetes-associated deaths in the old people and in winter. Conclusions This studyprovides evidences that NO2 and SO?2 in Nanjing are significantly associated with diabetes- related deaths.

Kinetics of serum HBsAg in CHB patients with nucleos(t)ide analogues treatment

2017, 37(6):  817-820. 

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Objective To summarize and analyze the dynamic change of HBsAg levels in patients with chronic Hepatitis B (CHB) after receiving nucleos(t)ide analogues (NAs) as antiviral treatment. Methods Patients who were performed quantitative Hepatitis B surface antigen(qHBsAg) from July 30, 2012 to December 30 ,2016 in Peking Union Medical College Hospital were retrospectively enrolled. QHBsAg, HBV DNA, quantitative HBeAg were collected and analyzed at baseline and at 192-week follow-up every 24 weeks. QHBsAg and quantitative HBeAg were assessed with CMIA .HBV DNA were assessed with PCR and COBAS Amplicor. Results 60 patients were included. Patients in HBeAg-positive group had higher HBV DNA than that in HBeAg-negative group （P＜0.05）at baseline and the two groups both were under detection limit after 48 weeks.Baseline qHBsAg in HBeAg positive-group and negative-group were (3.43±0.73) log10IU/mL, (3.08±0.47) log10IU/mL respectively .QHBsAg in HBeAg-positive group was higher than that in HBeAg negative-group on all follow-ups（P＜0.05） except 48weeks.However on 168 weeks and 192 weeks , difference between the two groups was statistically significant（P＜0.05）. In HBeAg-positive group,quantitative HBeAg dropped significantly during antiviral treatment. Conclusion HBV replication can be suppressed in the process of long-term NAs treatment in CHB patients.However qHBsAg decline is not so obvious, which indicates that HBsAg loss in the cure target is difficult,and long-term NAs therapy is still essential.

Effects of sulfur lamp, heat radiation lamp, fluorescent lamp and LED lamp lighting on reproductive system in depressive male rats

2017, 37(6):  821-827. 

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Objective To investigate the effects of different lighting(sulfur lamp, heat radiation lamp, fluorescent lamp and LED lamp) on reproductive system in depressive male rats. Methods CUMS depressive model of rats in vivo was established through the stimulations of chronic unpredictable mild stress(CUMS), then they were randomly divided into 5 groups：depressive model group, sulfur lamp group, heat radiation lamp group, fluorescent lamp group and LED lamp group. After 45 d of continuous illumination, serum was collected and reproductive organs were removed from rats, then the organ index of testicle，epididymis，seminal vesicle and kidney was calculated, the concentrations of testosterone (T), estradiol (E2), prolactin (PRL) and progesterone (PROG)in the serum were detected by ELISA, the histomorphological lesion of testicle was observed under microscope with hematoxylin-eosin (HE) staining , the expression of melatonin receptors (Mel 1a) , 3β-HSD and P450scc of testicle were detected by Western blot. Results The organ index of testicle，epididymis and seminal vesicle, the concentration of T, E2, PRL and PROG, and the expression of Mel 1a, 3β-HSD and P450scc in the depressive rats were significantly lower than that of the controls(P<0.05), and the seminiferous tubules of the testis of depressive rats were atrophied. However, the level of T, E2, PRL and PROG were increased，the cell morphology of the testicle was improved, and the expression of Mel 1a, 3β-HSD and and P450scc were upregulated after the sulfur lamp lighting in the depressive male rats compared with depressive model group (P<0.05). Conclusions The testis structure and functions of depressive male rats are improved by sulfur lamp lighting.

A systematic review of protective ventilation for surgery in the prone position

2017, 37(6):  828-833. 

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Objective To evaluate the application ofprotective ventilation(PV) versus conventional ventilation(CV)for surgery in the prone position. Methods We searchedPubMed, Embase, the Cochrane Library, WanFang Dataand other Chinese databases to collect the randomized controlled trails (RCTs) on intraoperative PV in comparison with CVfor surgery in the prone position.Two authors independently identified the studies, performed data extraction and assessed the risks of bias in the included studiesaccording tothe Cochrane Handbook for Systematic Reviews of Interventions. The reviewers conducted data analyses with RevMan software. Results A total of 9 RCTs involving 449participants were included.The resultsshowed that the incidence of postoperative pulmonary complications (RR 0.30, 95% CI 0.12~0.73, P<0.01) and peak pressure (MD -3.52, 95% CI -6.93~-0.11, P<0.05) were lower in the PV group. Intraoperative PaO2/FiO2was higher (MD 37.28, 95% CI 22.67~51.89, P<0.001) and alveolar-arterial oxygen difference was lower(MD -45.50, 95% CI -61.35~-29.65, P<0.001) in the PV group. Conclusions Application of low tidal volume ventilationin combination withpositiveend-expiratory pressure(with or without recruitment maneuver)decreases postoperative pulmonary complications, reduces peak pressure and improves oxygenationfor surgery in the prone position.

Effect of Roux-en-Y gastric bypass surgery on renal gluconeogenesis in obese rats

2017, 37(6):  834-838. 

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Objective To investigate the effect of Roux-en-Y gastric bypass surgery on renal gluconeogenesis, and to explore the mechanism of improving glucose control and insulin sensitivity in obese rats. Methods Rats were randomly divided into the obese group (n=100) consumed high-fat diet (HFD) or the normal control group (NC group, n=20) consumed standardized diet. Top 30% of animals fed HFD by change in body mass were subdivided into three groups: obese control group (OB group, n=10), obese under RYGB group (RYGB group, n=10) or sham-operation (Sham group, n=10). Intraperitoneal glucose tolerance tests (IPGTT) were performed meanwhile the AUC of blood glucose concentration-time after surgery. Fasting glucose, insulin and lipid were measured respectively and HOMA-IR were calculated. The mRNA levels and protein levels of the key enzymes (G6P and PEPCK) of renal gluconeogenesis were examined with RT-PCR and Western blot respectively. Results Compared with OB group and Sham group, the level of body mass, lipid, blood glucose, insulin, HOMA-IR and AUG were significantly decreased (P<0.05), the expression of G6P and PEPCK mRNA and protein were significantly reduced (P<0.05) in RYGB group. Conclusions Roux-en-Y gastric bypass surgery can improve glucose control and insulin resistance, which may be related to decrease levels of mRNA and protein of G6P and PEPCK, weaken gluconeogenesis and glucose output in renal cortex.

Tetramethylpyrazine inhibits LPS-induced inflammation of human type II alveolar epithelial cells

2017, 37(6):  839-844. 

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Objective To study the protective effect of TMP(Tetramethylpyrazine) on LPS(Lipopolysaccharides )-induced inflammatory response of human type II alveolar epithelial cells (HAECII) and its corresponding mechanism in vitro. Methods HAECII (A549 cells derived from human lung adenocarcinoma cells) were cultured in vitro. Inflammation models were established using A549 cells after LPS stimulation. TMP and FK866 (a specific inhibitor for pre-B cell colony-enhancing factor), were added to intervene respectively. Expression levels regarding mRNA and protein of inflammatory factors, including tumor necrosis factor-α (TNF-α), interleukin-1β(IL-1β), interleukin-8 (IL-8) and PBEF(pre-B cell colony-enhancing factor) were detected by q-PCR and Western blot, respectively. The activation of NF-κB(Nuclear factor κB) was monitored using Western blot by measuring the changes in phosphorylated P65 protein levels in both nucleus and cytoplasm. Results Both the mRNA and protein levels of TNF-α, IL-1β, IL-8 and PBEF in A549 cells were significantly higher after LPS stimulation than those in the control group(P<0.001). Meanwhile, the phosphorylation of P65 protein in the nucleus and cytoplas was higher(P<0.001). The expression of the aforementioned inflammatory factors and the phosphorylation of P65 protein were significantly lower after TMP intervention than those of LPS group(P<0.05). In comparison, after FK866 was added, the expression of TNF-α, IL-1β and IL-8 and the phosphorylation of P65 protein were also decreased(P<0.01). Conclusions TMP may be involved in the reduction of PBEF expression, which therefore inhibits NF-κB activation, antagonizes alveolar epithelial cell inflammatory response.

Effect of silenced/enhanced Stat1 gene on proliferation and invasion in pancreatic cancer cells

2017, 37(6):  845-848. 

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Objective To investigate the proliferation and invasion in human pancreatic cancer after Stat1 gene silenced or enhanced and to explore the possible underlying mechanisms．Methods BxPC-3 cells were divided to three groups: Stat1 group which transfect with specific Stat1 plasmid, Si Stat1 group which transfected with specific Stat1 small interfering RNA (siRNA), and control group. Western blot was used to detect the expression of Stat1, VEGF, MMP-2 and MMP-9 in BxPC-3 cells. MTT method was used to detect the proliferation of BxPC-3 cells. Transwell was used to detect the invasion of BxPC-3 cells．Results The proliferation and invasion were significantly enhanced in Pancreatic Cancer Cells after Stat1 gene silenced, and the expression of VEGF, MMP-2 and MMP-9 were increased. The proliferation and invasion were significantly decreased in Pancreatic Cancer Cells after Stat1 gene enhanced． Conclusion Stat1 inhibits the proliferation and invasion of pancreatic cancer. VEGF，MMP-2 and MMP-9 possibly play a role in this process.

Inhibitory effect of decorin on invasion of colorectal adenocarcinoma cell HCT116 in vitro

2017, 37(6):  849-854. 

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Objective To investigate the effect and mechanism of decorin (DCN) on invasion of colorectal cancer cell HCT116 in vitro. Methods Transwell assay was employed to detect the invasion of HCT116 cells; Real-time PCR was used to detect the expression of CD133 and TIMP-2 mRNA of HCT116 cells; Western blot method was used to detect the expression of HIF-1α, CD133 and TIMP-2 protein of HCT116 cells. Results 1) When the concentrations of DCN is 0,1and3mg/L,under the conditions of normal oxygen and hypoxia, the number of invasive cells wererespectively(241±46),(168±46),(51±17)fields in each well(P<0.01) and(207±61),(213±64),(156±54),(44±17)fields in each well(P<0.01). 2) Under the normoxic conditions, the TIMP-2 mRNA and protein in HCT116 cells were increased by DCN(3mg/L)(P<0.01), but that of CD133 were not affected. 3) DCN(3mg/L) significantly decreased the expression of HIF-1α/CD133/TIMP-2 protein in HCT116 cells under hypoxia(P<0.01), but had no significant effect on the expression of CD133mRNA. Conclusions Under the conditions of hypoxia and normal oxygen,DCN may exert through different mechanisms toinhibit the invasion of colorectal adenocarcinoma cell line HCT116 in vitro.

Effects of fasted, diabetes and obese conditions on mouse hepatic SOCS2 gene expression

2017, 37(6):  855-859. 

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Objective To determine the expression levels of SOCS2 in the mouse livers under fasted, diabetes and obese conditions and to study the effect of SOCS2 on gluconeogenesis. Methods Animal were divided into 3 groups: C57BL/6J mice, the control group was fed ad libtum and the experimental group was fasted for 24 h. Diabetes model db/db and the control db/m mice were fed ad libitum. Obese model ob/ob and the control C57BL/6J mice were fed ad libitum. All the mice above were sacrificed and total RNA was isolated from mouse livers and reverse transcribed to cDNA. The expression levels of SOCS2 and gluconeogenesis genes in the mouse livers in the 3 groups above were detected by real-time quantitative PCR. SOCS2 was overexpressed in the primary C57BL/6J mouse hepatocytes by the adenovirus system. The effect of SOCS2 on glucose production was measured by glucose output assay. Results C57BL/6J mouse hepatic SOCS2 expression was suppressed by fasted status. The expression levels of SOCS2 were decreased in the livers of db/db and ob/ob mice. In contrast, the key regulators of gluconeogenesis, PGC-1α, PEPCK and G6Pase exhibited the opposite expression pattern as SOCS2 in the livers under identical fasted, diabetes and obese conditions. The protein was Mr 23 000 and glucose production was inhibited after SOCS2 was overexpressed in the primary C57BL/6J mouse hepatocytes by adenovirus system. Conclusions SOCS2 is speculated to inhibit gluconeogenesis in the C57BL/6J mouse primary hepatocytes, and SOCS2 may be a new target for the treatment of type II diabetes.

Berberine regulates gut microbiota and Th17/Treg balance in sleep deprived rats

2017, 37(6):  860-864. 

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Objective To investigate the effect of berberine on gut microbiota and T helper cell 17 (Th17), regulatory T cell (Treg) cell in sleep deprived rats. Methods SD rats were randomized into control group, model group, low-dose and high-dose group (BBR1 and BBR2, 100 mg/kg and 200 mg/kg, administrated orally). Sleep deprived rat model was established by the small-platforms-over-water method. The number of bacteria in rectum content of rat was detected. The ratio of Th17/Treg was evaluated by flow cytometry. The expression of interleukin 17 (IL-17), RAR-related orphan receptor (ROR) C, and Forkhead Box Protein P3 (Foxp3) mRNA was evaluated. Results The amount of Clostridium perfringens was elevated (P<0.05), the amount of other microbiota decreased (P<0.05), Th17/Treg ratio increased (P<0.05), IL-17 and RORC expression enhanced (P<0.05), Foxp3 expression decreased (P<0.05) in the gut of model rats. In contrast, treatment with berberine inhibited the amount of Clostridium perfringens (P<0.05), promoted the amount of other microbiota (P<0.05), dampened Th17/Treg ratio (P<0.05), regressed IL-17 and RORC expression, augmented Foxp3 expression. Conclusions Various doses of berberine can counteract gut microbiota imbalance and Th17/Treg imbalance induced by sleep deprivation.

Outcome evaluation of clinical pharmacists intervention on clinical antimicrobial agents

2017, 37(6):  865-869. 

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Objective To investigate the effects of clinical pharmacist intervention in the clinical application of antibiotics combined with the index management. Methods The data of antibiotics usage rate and intensity, the microbial test sample submission rate, and the clean incision preventive usage rate of inpatients and outpatients were analyzed by chi-square test in the same period before (2015) and after (2016) the clinical pharmacist intervention. Results After intervention, the antibiotics usage rate of inpatients and outpatients were lower than that of 2015 (50.02% and 25.72%, P<0.05). In 2016, the usage intensity of antibiotics was 50.59%, while it was 62.39% in 2015. At the same time, the microbial test sample submission rate increased from 13.96% to 27.49% (P<0.05).In 2016, the clean operation preventive usage rate of antibiotics decreased month by month. In the meantime, the reasonable rate of delivery timing, variety selection, and administration time of clean operation preventive usage of antibiotics were on the rise. Conclusion The professional intervention of clinical pharmacist is benefit for the routine, standardization, and professionalization of clinical antibiotics application.It also promotes the rational use of drugs and the progress of hospital pharmacy.

Current understanding of ABCA1 in lipid accumulation

2017, 37(6):  870-873. 

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ATP-binding cassette transporter A1 (ABCA1) is an membrane protein,using ATP for transferring substances,modulated by liver X receptors(LXRs), retinoic X receptors(RXRs), sterol regulatory element binding proteins(SREBPs), microRNAs and other upstream modulators. ABCA1 plays an important role in multiple pathways related to lipid accumulation. Upregulating ABCA1 expression can reduce lipid accumulation, hence offers a promising strategy for metabolic diseases.

Research progress of robotic surgery simulation training platform in urology

2017, 37(6):  874-879. 

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As the development of robotic surgery in urology, the simulation training platform is playing an important role in the training of urologists on robotic surgery training. Currently, the available training simulators for urological robotic surgery are composed of virtual simulators such as VR and AR, and physical (mechanical) simulators such as dry-lab and wet-lab. The reality and validity of various simulation training platforms have been validated in different degree by many researches, which demonstrated the useful and helpful effect on the training and improving of robotic surgery skills. However, we need further study to develop more useful and precise training modules, to integrate advantages of different simulators, and to establish more reasonable learning curve.

Progress in neural stem cells therapy for traumatic brain injury

Xiao ZHANG Jun-ji WEI, Ren-zhi WANG,

2017, 37(6):  880-883. 

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The neural stem cells (NSCs) can migrate into the injured areaand differentiate into neurons or oligodendrocytes. Endogenous neurogenesis could potentially be harnessed as a putative therapyfor neural injury. But the complexmicro-environment due to TBI will be one of the biggest challenges for endogenous NSCs to performneural regenerations.Exogenous NSCs have been shown to be able to survive in host tissues and regulate microenvironment via paracrine effects. Thus, transplantation of NSCs to assist neural regeneration has become an attractive option. Recently,rapid advances in the stem cell biology have raised appealing possibilities of replacing damaged or lost neural cells by transplantation of in vitro-expanded stem cells and/or their neuronal progeny.

Research progress of capecitabine combined chemotherapy with biweekly scheme

2017, 37(6):  884-888. 

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As an oral chemotherapy drug, capecitabine is safe, effective and easy to use, has been widely used in esophageal cancer, gastric cancer, colorectal cancer, breast cancer, ovarian cancer and other solid tumor. The chemotherapy scheme mainly for 3 weeks, each cycle of capecitabine for 14 days, and 7 days off. As a modification to conventional chemotherapy, the biweekly scheme with capecitabine showed low toxicity and well tolerated by patients, short hospitalization time and other characteristics, it is worth of application in clinical.

Application of multivariate generalizability theory in the study on the reliability of objective structured clinical examination

2017, 37(6):  889-893. 

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Objective To evaluate the reliability of objective structured clinical examinationbased on standardized patients in clinical skill test for medical students. Methods To analyze the scores of 8 stations OSCE by applying multivariate generalizability theory. Results Therelative G coefficient of the 8 stations OSCE was 0.886, and the absolute G coefficient was0.883. Conclusions Multivariate generalizability theory is an objectively and scientifically toll to evaluate theOSCE stations, and the analysis results are helpful to improve the practice for future OSCE.

Development and application of morphological multimedia website based on autonomous learning

2017, 37(6):  894-898. 

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Objective In view of the trend of networking development in modern high education and the characteristics of students’ strong self-learning ability, Peking Union Medical College established a multimedia morphological teaching website from 2013 including human anatomy, neuroanatomy, histology and embryology. Methods According to the teaching demand, the use of the ASP script, combined with Mysql database completed the website development, from the interface design to the curriculum, the syllabus, presentations and laboratory videos uploading. Results Through the questionnaire survey, 45% of the students use website more than 3 times a week, and “course content” column has the highest use frequency (79%). An independent learning platform effect has been achieved. Conclusions After nearly 4 years exploration and practice, multimedia website has become an important part of morphological courses, as a kind of new teaching mode, not only popular for college teachers and students, but also widely used in clinical teaching.

Comparison between Chinese and American physiology textbooks

Li YAN Jing WANG

2017, 37(6):  899-902. 

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Textbooks are the base of teaching. The physiology textbooks used in higher medical education was compared between China and the United States. Guyton and Hall Textbook of Medical Physiology was chosen as the major representative of the United States. The overall framework, the selection and arrangement of the contents, the application of tables and pictures, and the relationship between basic medicine and clinical medicine were discussed. It is hopefully that this paper would provide some reference for reform in specialized course system and teaching contents of physiology, especially for improving the constructive quality of textbooks in the future.

Effects of CBL in teaching human embryology for medical students

2017, 37(6):  903-906. 

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Object To investigate the effects of CBL in teaching human embryology for medical students. Methods After having seminar with clinical case, scoring rate of questions discussed and not discussed in final exam were compared in recent 3 years. Embryological knowledge were evaluated in 1, 2, 3 years after embryology class by online test. Results There was no difference of total scoring rate among 3 grades in final exam, while questions discussed was higher significantly than those not discussed（p<0.01）. Scoring rate of objective questions discussed was higher significantly than those not discussed. Scoring rate of online test decreased with years after embryology class, while questions discussed were better than not discussed. Conclusions Seminar with clinical case as CBL could improve the learning outcomes of embryology in short and long term memory.