Table of Content

05 May 2012, Volume 32 Issue 5

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Old Topic of Conversation and New Questions: Studies on Tumor Metablism

2012, 32(5):  461-461. 

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Role of Pyruvate Kinase M2 in tumor metabolism and development

2012, 32(5):  462-467. 

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Tumor cells always undergo glycolysis to metabolize glucose even in the presence of oxygen, which is termed aerobic glycolysis, also known as Warburg effect. A key sensor regulator of this process is pyruvate kinase M2 (PKM2), which catalyzed the PEP to pyruvate. It is particularly expressed in proliferative cells especially in tumor cells. It is always switched between a highly active tetrameric form and a nearly low active dimeric form, which determined whether the glucose carbons are converted to pyruvate under the production of energy or participated into synthetic processes. In tumor cells, the dimeric form of PKM2 is predominant. The amounts of Tumor M2-PK can serve as a biomarker to be detected in EDTA-plasma and stool samples of patients with various types of cancers. PKM2 is a pTyr-binding protein; it can be regulated by a lot of transcription factors such as HIF1α, and metabolic intermediates such as FBP. The crucial role of PKM2 in tumor glucose metabolism and proliferation makes it an interesting therapeutic target for the treatment of human cancers.

Lipopolysaccharide induces HMGB1 expression in RAW264.7 cells through p38MAPK-CBP signaling pathway

2012, 32(5):  474-480. 

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Objective To explore the signaling molecules relate HMGB1 expression in murine macrophage-like cell line RAW264.7 cells induced by lipopolysaccharide, HMGB1, p38MAPK, NF-κB and CBP were detected. Methods After stimulated by LPS, p38MAPK, NF-κB and CBP protein in cytoplasm and nucelus were observed by immunocytochemistry and laser confocal scanning microscopy. HMGB1 protein in supernatant was measured by ELISA, the expression of HMGB1 mRNA in cultured cells was determined by Real-time PCR. Cultured cells cytoplasm and nucleus HMGB1 protein were detected by Western blot. Results With LPS stimulation, the green fluorescence of p38MAPK was gradually increased and the green fluorescence of NF-κB was gradually weakened in cytoplasm, while in nucleus, the green fluorescence of both NF-κB and CBP was gradually increased, and reached the peak after 6 h. HMGB1 protein in supernatant and cytoplasm increased gradually from 12 to 48 h after stimulated by LPS. HMGB1 protein in nucleus decreased gradually from 12 to 24 h after LPS stimulated, increased gradually after 36 h(P<0.01). The expression of HMGB1 mRNA has no significantly changed from 0 to 12 h but enhanced significantly from 24 to 48 h after LPS stimulated (P<0.01). Conclusions LPS activated p38MAPK、NF-κB and CBP signaling pathway, which led to HMGB1 deacetylated and transferred into cytoplasm, finally released into excellular.

Senescence induced by ginsenoside Rg1 on human leukemia K562 cells

2012, 32(5):  481-486. 

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Abstract: Objective To observe the senescence effects and mechanisms of leukemia K562 cell line induced by ginsenoside Rg1 (Rg1). Methods The related mechanism was observed via inducing k562 cells senescence by optimal drug concentration and interaction time detected by MTT colorimetric test. Effect of Rg1 on cell cycle was analyzed using flow cytometry. The positive cells were detected by SA-β-Gal staining. The colony-formed ability were detected by Colony-Assay. The telomere lengths were detected by Southern blotting. The expression of proteins were detected by Western blotting and the ultrastructure changes were observed by transmission electron microscopy. Results Rg1 significantly inhibited the proliferation of K562 cells in vitro and arrested the cells in G2/M phase. The percentage of positive cells stained by SA-β-Gal was dramatically increased （P＜0.05）and the colony-formed ability in experimental group has been weakened（P＜0.05）. Senescence-related proteins P53、P21、P16、RB in experimental group were up-regulated（P＜0.05）. Telomere lengths became shorter. The observation of ultrastructure showed that cell volume increase, heterochromatin condensation and fragmentation, mitochondrial volume increase, lysosomes increase in size and number. Conclusions Rg1 could induce the senescence of leukemia cell line K562 and P53-P21-Rb, P16-Rb cell signaling pathway play a significant role in this process.

Effects of adeno-associated virus-mediated klotho gene delivery on the expression of runx2 and mmp-13 gene in the bone of the ovariectomy rats

2012, 32(5):  487-492. 

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Abstract: Objective To research on the effect of the recombinant adeno-associated virus vector containing klotho gene delivery to regulate for ovariectomized rats.Methods Female SD rats were randomly divided into sham operation group (S group) and model group.Model was successfully constructed with ovariectomy after 12 weeks,they were randomly divided into model group (O group), 17β-estradiol (E group), klotho gene group (KO group), empty vector group (GO group), all were sacrificed after 12 weeks.Bone mineral density(BMD) of the femurs and tibia were measured. The fluorescent expression of renal klotho was observed by Cryo-sectioning Technique.The Runx2 and MMP-13 mRNA expression of bone tissue were detected by reverse transcription-polymerase chain reaction(RT-PCR).Expression of klotho protein in kidney and Runx2,MMP-13 protein in bone were detected by immunohistochemistry. Bone morphological changes of the different groups were observed by HE staining.Results BMD of KO and E groups were significantly higher than those in the O and GO groups(P<0.05). Specific expression of mouse klotho was seen only in KO group.Renal klotho protein in KO group were increased obviously by immunohistochemistry.Compared to O group,the expression of Runx2 mRNA increased greatly,but the expression of MMP-13 mRNA decreased in bone tissue of KO group (P<0.05). Immunohistochemistry analysis showed,the average absorbance of Runx2 was 411±96 in KO group,it was a significant higher than 353±50 of O group(P<0.05). The average absorbance of MMP-13 was 397±84 in KO group,the value of O group was 656±89,so KO group showed a significantly lower than those in O group (P<0.05).The trabecular bones in KO,E,S groups tightly packed, interconnected to form a network,had relatively complete histologic structure.Conclusion Klotho gene delivery attenuates the progression of osteoporosis and bone microstructure damage in ovariectomized rats, these results suggest that klotho gene may play a role in the progression of osteoporosis.

Effect of nanosecond pulsed electric fields on treatment of human melanoma A375 cell xenograft in nude mice

2012, 32(5):  493-499. 

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ABSTRACT Objective：To investigate the effect of nanosecond pulsed electric fields(nsPEF)on human melanoma A375 cell xenograft in nude mice and its possible mechanisms. Methods：Nude mice models were randomly divided into long-term treatment group for observing tumor growth and survival time （ ten mice every group），and short-term treatment group (includes 2h group、4d group and control group,4、6 and 6 mice ,respectively) exposed to the 40kV/cm, 200ns, 1HZ, 1000 pulses .Tumor morphology was observed with HE staining; The cell apoptosis was detected by DNA agarose gel electrophoresis and terminaI deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay; The expression of Bax and Bcl-2 protein was detected by Western blot and SP immunohistochemistry. Results: Tumor turned grey without bleeding spots minutes after long-term treatment,the tumor volume was significantly smaller in treatment group than in control group ( P<0.01),the mean survival time was significantly longer in treatment group()than in control group()(P<0.01).Cell necrosis was found in 4d group;DNA ladder was observed in 4d group;the apoptosis rate of tumor cells was significantly higher in 4d group than control group ( P<0.01);the expression of Bax was significantly higher in 4d group than in control group (P<0.01); the expression of Bcl-2 was significantly lower in 4d group than in control group after short-term treatment( P<0.01). Conclusion: nsPEF may cause melanomas complete remission without recurrence by inducing cell apoptosis under the Bax and Bcl-2 gene regulation mechanism .

Up-regulation of microRNA-125b in gastric cancer and its roles in promoting cell proliferation

2012, 32(5):  500-504. 

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Objective To evaluate the expression level of microRNA-125b (miR-125b) in gastric cancers and study its roles in cell proliferation and apoptosis. Methods The expression of miR-125b was detectable by real-time PCR in 40 gastric cancer tissues and paired cancer-adjacent tissues. The synthesized miR-125b mimic was transfected into cultured gastric cancer cell lines HGC-27 and MGC-803. The cell proliferation and apoptosis were measured by CCK-8 and flow cytometry respectively. Results MiR-125b was significantly up-regulated in gastric cancer tissues compared with cancer-adjacent controls（P<0.01）. Ectopic expression of miR-125b in HGC-27 and MGC-803 cells increased cell proliferation, whereas decreased cell apoptosis. Cells transfected for 72h, HGC-27（scramble：1.632±0.09，mimic：2.473±0.08），MGC-803（scramble：1.603±0.05，mimic：2.554±0.07）），Conclusion MiR-125b may act as an oncogene in gastric cancer via interfering with cell proliferation and apoptosis.

Influence of P.yoelii MIF on mouse BM-DC

2012, 32(5):  505-509. 

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Abstract: Objective Detect the role of P.yoelii plasmodium derived macrophage migration inhibitory factor (PyMIF) recombination protein on the phenotype and function of bone marrow dendritic cells (BM-DC). Methods : BM-DC induced from bone marrow precursors were treated with PyMIF or MmMIF，then cell surface molecular such as TLR2、TLR4、CD80、CD86、CD40、MHCII were detected by flow cytometry and IL-12、IL-10 were analyzed by ELISA. PyMIF or MmMIF treated BM-DC were first stimulated by LPS，then co-cultured with CD4+T or CD8+T cells, CD69 and IL-2 were detected as previously described to indentify the activity of T cells. What’s more, the tytotoxic action of CD8+T was also measured. Results: PyMIF decreased the TLR4 expression of BM-DC，but had neither effect on TLR2、CD80、CD86、CD40 and MHCII expression, nor influence on IL12、IL-10 secreting. What is more, PyMIF down-regulated CD69 expression of CD8+T through BM-DC, and could not influence CD69 expression and IL-2 secrecting of CD4+T and IL-2 secrecting of CD8+T cells through BM-DC. Conclusion：PyMIF could down-regulated TLR4 expression of BM-DC to prevent being attacked by immune system, so as to survive in the organism.

MiR-21 modulates the invasiveness of QBC939 cells through negatively regulating RECK expression

2012, 32(5):  510-515. 

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Objective To investigate the expression profile of miR-21 in human cholangiocarcinoma tissues and QBC939 cell line and probe the function of miR-21 in cholangiocarcinogenesis. Methods MiR-21 expression in human cholangiocarcinoma tissues and QBC939 cell line is measured by using Real-Time PCR and Northern Blot, respectively. Cell growth and apoptosis was analyzed in QBC939 after transfected with Anti-miR-21. Specific target analysis is performed by using dual-reporter gene assay and FACS. In Vitro invasion assay is performed to probe the effect of miR-21 on QBC939 invasiveness. Results Expression analysis reveals that miR-21 levels depicted a significant up-regulation as compared to the matched normal bile duct and miR-21 levels are augmented approximately 3.4-fold in QBC939 cells. Silencing of miR-21 in QBC939 by using anti-miR-21 decreases cell growth and induces cell apoptosis. RECK is identified as a direct effector of miR-21 and miR-21 promotes QBC939 cell invasion in vitro through negatively regulating miR-21-RECK program. Conclusion Increased miR-21 expression is a frequent event in human cholangiocarcinoma. Augmented miR-21 promotes cell growth and dampens cell apoptosis. The invasiveness is enhanced by miR-21 through inhibiting RECK expression.

Roles of ULBPs and MIC molecules in the diagnosis and evaluation of sub-health status

2012, 32(5):  516-519. 

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Objective Establish a database of ULBPs and MICA / B molecule express level in peripheral blood of sub-health state population, and provide a reference for evaluation the health of overall population and prevention of diseases. Methods The health statuses of volunteers were evaluated according to physical examination and questionnaire survey. After that, their serum protein levels of related molecules were detected by ELISA. Results The average concentrations of MICA, MICB, ULBP1 and ULBP2 in serum were higher in sub-health population compared with the healthy group. However, the levels of ULBP3, ULBP4 and ULBP5 were lower than those in healthy group. Univariate analysis showed that only MICB has significant difference (P <0.01) between the sub-healthy group and healthy group. Multivariate logistic regression analysis also confirmed this result. In addition, serum levels of MICA and ULBP3 in male was statistically significant higher than those in female (P<0.05). Additionally, ULBP2 and ULBP4 increase with age. Conclusions Together with other immune parameters, serum levels of soluble MICB may be useful in the diagnosis of sub-health.

The effects of bone marrow derived mesenchymal stem cells on the tumor formed by HT-29 cells

2012, 32(5):  520-524. 

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Objective To study the effects of bone marrow derived mesenchymal stem cells on the growth and differentiation of the tumor formed by HT-29 cells. Methods In vivo, nude mice were divided into 3 groups— HT-29 group, BMSC group and HT-29+BMSC group, in which cell suspensions were subcutaneously injected into nude mice. Tumors were removed after 7 weeks .The volumes and weights of the tumors were measured and HE staining was performed. The expression of an epithelia marker —E-CADHERIN was detected using RT-PCR and immunohistochemistry staining. In vitro, HT-29 cells were co-cultured with BMSCs, the morphology of HT-29 cells was observed, and the expression of EMT related genes were detected using RT-PCR. Results In vivo, no tumor was observed in the BMSC group, and no significant difference was found in the volume and weight of tumors between the HT-29+BMSC group and the HT-29 group. But the tumor cells in the HT-29+BMSC group showed poorly differentiated and lower level expressed of E-CADHERIN compared with the HT-29 group. In vitro, after coculturing with BMSCs, morphological changes of HT-29 cells were observed and were consistent with the expression of EMT related genes. Conclusion The differentiation degree of HT-29 cells is correlated to EMT induced by BMSCs microenvironment.

Construction and expression of anti-hγδTCR-ScFv and analysis of its biological functions

jing zheng

2012, 32(5):  525-532. 

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Objective To construct and express the single chain variable fragment ( Fv) of a monoclonal antibody against human γδ TCR which can stimulate proliferation of human γδ T cells．Methods The genes of the heavy (VH) and light chains (VL) of antibody were cloned by RT-PCR from a hybfidoma cell line G5-4，which secreted the monoclonal antibody (mAb) against human γδ TCR．Overlapping extension PCR was used to connect the VH and VL genes with a short linker encoding (Gly4Ser)3 to construct the anti-hγδTCR-ScFv gene G5-4ScFv. Recombinantplasmid pET22b(＋)-G5-4ScFv was constructed by inserting G5-4ScFv fragment into the prokaryotic expression vector pET22b(＋). The anti-hγδTCR-ScFv (G5-4ScFv) was expressed in E.coli TransB(DE3) by induction with IPTG and purified with Nickel-affinity chromatography column. Then G5-4ScFv was analyzed by SDS-PAGE and Western-blot assay. Its binding ability to human γδ T cells was measured by flow cytometry. The purity and biological characteristics including cytokine secretion and cytotoxicity of γδ T cells expanded by G5-4ScFv stimulation, were detected by flow cytometry and LDH methods．Results G5 -4ScFv showed a molecular weight of 30ku in SDS-PAGE. G5-4ScFv binded to γδ T cells and greatly inhibited the binding of a pan-γδTCR antibody with γδ T cells. The purity of γδ T cells expanded by G5-4ScFv stimulation was up to 90% after culture of 2 weeks. These γδ T cells exhibited IFN-γ and TNF-α secretion ability and strong cytotoxicity against Daudi cells. Conclusion Anti-hγδTCR-ScFv with biological functions has been successfully constructed and expressed in prokaryotic expression system, which provides the basis for further studying on its potential application in tumor immunotherapy.

Effects of lentivirus-mediated GDNF on biologic characteristics of cynomolgus monkey mesenchymal stem cells

2012, 32(5):  533-538. 

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Objective To explore the effects of lentivirus-mediated GDNF on biologic characteristics of cynomolgus monkey mesenchymal stem cells. Methods Under sterile condition, bone marrow mononuclear cells of cynomolgus monkey were isolated by density-gradient centrifugation and MSCs were cultured in vitro. GDNF lentivirus infected cynomolgus monkey MSCs (cMSCs). Using ELISA, Real-time PCR, flow cytometry (FCM), BrdU incorporation and other experimental methods, GDNF gene expression, cell surface markers, proliferation and differentiation of cynomolgus monkey MSCs were observed before and after the infection of lentivirus. Results cMSCs highly expressed GDNF in vitro after infection of lentivirus. After the infection of GDNF lentivirus, cell morphology and adipocyte differentiation of cMSCs did not change significantly, but cell surface markers CD34, CD90 and Stro-1 positive cells increased in vitro, and the proliferative potential decreased. Conclusion The cell surface markers and the proliferative potential of cMSCs changed after the infection of GDNF lentivirus, but the adipocyte differentiation ability of MSCs had no effect.

Involvement of Increased Expression of C3G in Myocardium Around the Infarcted Zones in the Acceleration of Post-infarction Cardiac Remodeling Induced by Isoproterenol in Rats

2012, 32(5):  539-543. 

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Objective To address the involvement of C3G (CrkSH3－binding guanine nucleotide exchange factor) in the acceleration of post-infarction cardiac remodeling induced by isoproterenol (ISO), C3G protein expression was investigated in myocardium around the infarcted zones in rats with ISO stimulation or not. Methods Myocardial infarction (MI) and sham-operation models were established by Litwin’s method. Seven days after operation, surviving rats were divided into MI, Sham, MI+ISO and Sham+ISO groups respectively, and were treated with physiological buffered saline 5ml/kg or ISO 5mg/kg intraperitoneally once every 3 days for 12 weeks respectively. C3G protein expression in myocardium around the infarcted zones was detected by Western blot. Results Twelve weeks after treatment, the normalized integral optical density (IOD) of C3G protein expression was (1.14±0.29, n=8) in the MI group, (0.90±0.10，n=6) in the Sham group, (1.51±0.18，n=10) in the MI+ISO group and (0.97±0.26, n=8) in the Sham+ISO group respectively. The C3G protein expression in myocardium around the infarcted zones was significantly increased in the MI group compared with the Sham group, and in the MI+ISO group compared with the Sham+ISO group, and in the MI+ISO group compared with the MI group respectively (All P<0.05). Conclusions C3G protein expression was significantly increased in myocardium around the infarcted zones. Moreover, ISO can further up-regulate its expressions in myocardium around the infarcted zones. The increase of C3G protein expression is involved in the post-infarction cardiac remodeling, ischemic cardiomyopathy and heart failure; Furthermore, its further increased expression is also involved in the acceleration of post-infarction cardiac remodeling, ischemic cardiomyopathy and heart failure induced by ISO.

Expression and significance of vascular endothelial growth factor in breast tissue in patients with breast cancer

2012, 32(5):  544-547. 

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Abstract Objective: To explore the expression and clinical significance of vascular endothelial growth factor in breast tissue in obese breast cancer. Methods：Breast tissue samples from 45 cases with breast cancer, 37 cases of normal weight with breast cancer, the expression of VEGF mRNA and protein of breast tissue were detected by RT-PCR and immunohistochemistry in obese breast cancer group and normal weight breast cancer group,and correlation was analyzed between VEGF mRNA and protein. Results: Compared with normal weight breast cancer group, the expression of VEGF mRNA and protein of breast tissue in obese breast cancer group increased obviously(P<0.05), while the expression of VEGF mRNA and protein were positively correlated (r=0.785，P<0.05); the expression of VEGF in breast tissue in obese breast cancer was relevant with histological grade and 5 years′relcurrence and metastasis, regardless of age. Conclusions: Obese patients with breast cancer own a high expression trend of expression of VEGF mRNA and protein of breast tissue, which indicates that VEGF maybe play an important role in occurrence , evolvement and prognosis of obesity-related breast cancer.

Inhibition role of Artesunate on esophageal cancer by inducing cell apoptosis

2012, 32(5):  548-553. 

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[Abstract] Objective To research inducing apoptosis function of artesunate and explore the mechanism of inhibition effect of artesunate on esophageal cancer. Methods Cell apoptosis, cell cycle, bcl-2 and bax protein expression level of Eca109 cells after the treatment of various concentration of artesunate(Art) (0、10、20、40μg/ml) for 24h were detected by flow cytometry(FCM). Results Cell apoptosis of Eca109 cells was significantly increased after the treatment of Art for 24h (P<0.05). The Eca109 cell bcl-2 protein expression level and cell proliferation index(PI) of Art group were lower than control group (P<0.05), but the bax protein expression level was higher in a concentration-dependent manner. Conclusion Artesunate have the inhibition effect to esophageal cancer by regulating the bcl-2, bax protein expression level and cell proliferation so inducing cell apoptosis.

Effects of siRNA-Gli-1 combined BCNU on prIiferation，apoptosis in Glioma Cell line U251

2012, 32(5):  554-560. 

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Objective:To observe the inhibition effect and mechanism of siRNA-Gli-1 and BCNU on the expression of Hedgehog(Hh) signal pathway and the proliferation and aptosis in glioma 251 cell. Methods:　Five groups were divided : BCNU group( BCNU),siRNA-Gli-1 group (si RNA- Gli -1)、combination group（siRNA-Gli-1 + BCNU）、transfection reagents group (transfection reagents）and blank control group(RPMI1640). siRNA-Gli-1 was transfected into U251 glioma cells, the mRNA and protein expression of Gli-1 gene were assessed by real- time quantitative poly merase chain reaction ( RT-PCR) and western blotting. After being treated by BCNU and siRNA-Gli-1,the proliferation and apoptosis of U251cells were examined by MTT, Flow Cyto- metry , Annexin V methods . RT-PCR and Western blotting was used to detect the mRNA and protein expression of Bcl-2、CyclinD1 and Bax in Hh signal pathway. Results:　 The expression of Gli-1 in siRNA- Gli-1 and combination group declined distinctly,as compared with other three groups (P<0.05) ; Flow Cytometry analysis showed that the proliferation and apoptosis of U251 cells in BCNU group、siRNA- Gli-1 group and combination group was inhibited and increased respectively , and G0/G1 phase rate increased and S phase rate decreased, as compared with transfection reagents group and blank control group(P < 0. 05)；In siRNA-Gli-1 group and com- bination group , especially in combination group , the expression of Bcl-2 and CyclinD1 decrea- sed obviously and the expression of Bax increased or had no change(P < 0. 05). Conclusion　: Combine application of siRNA-Gli-1 and BCNU can enhance the effect of inhibition to U251cell , it may be related to the inhibitory to the expression of Gli-1,as well as Bcl-2 and CyclinD1 in Hh pathway ,and the expression Bax may be increased.This effect can be achieved by cell cycle arrest and decreasing the ratio of Bax/Bcl-2..

Expression of TPX2 in cervical carcinoma and significance

2012, 32(5):  561-565. 

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[ABSTRACT] Objective:To study the expression of TPX2 protein and mRNA in human cervical cancer and the relationship between its expression and clinicopathologyical features of cervical cancer.Methods:Immunohistochemistry、reverse transcription polymerase chain reaction were used to analyze the expression levels of TPX2 mRNA and protein respectively, in fresh invasive cervical squamous cancer (n=62)、cervical adenocarcinoma（n=17） normal cervical uteri (n=15) ,CIN tissues(n=30), squamous carcinoma of the cervix Siha cell lines and adenocarcinoma of the uterine cervix Hela cells，①By RT-PCR, there was no expression of TPX2 mRNA in normal cervical tissues, The expression of TPX2 mRNA cervical squamous carcinoma is 0.441 ± 0.011, cervical adenocarcinoma is 0.467±0.031. Hela cell lines is 1.923±0.04,Siha cell lines is 1.679±0.042，There was significant difference between them and normal cervical tissues, (P<0.05)②By immunohistochemistry, normal cervical tissues did not express TPX2. The expression of TPX2 in CIN and cervical carcinoma were increased respectively ,There was significant difference between the three tissues (P<0.05). normal cervical glandular epithelium tissues did not express TPX2，The expression of TPX2 in adenocarcinoma of the uterine cervix is strong positive, There was significant difference between the two tissues (P<0.01).③The expression of TPX2 were correlated with histologicalgrade、 FIGO stages、 and lymph nodemetastasis (P<0. 05) and not correlated with age (P>0.05) 。④The expression of TPX2 in cervical gland cancer Hela cell line is stronger than squamous cancer Siha cell line. Conclusion:There is no expression of TPX2 in normal cervical tissues. TPX2 is probably involved in carcinogenesis and development of cervical carcinoma.The detection of TPX2 expression may play an important role in early diagnosis and evaluation of prognosis in patients with cervical carcinoma.The high-expression of TPX2 might become a novel biomarker for studying them echanism underlying the tumorigenesis and lymphaticmetastasis of cervical cancer and therapeutics target point for cervical cancers.

Increased Expression of c-kit in ICC in Colon of Rat with Visceral Hypersensitivity

2012, 32(5):  566-569. 

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Abstract: Objective Through research of the expression of c-kit in interstitial cells of cajal(ICC) in colon of rat with visceral hypersensitivity, to investigate the role of ICC in the pathophysiology of visceral hypersensitivity.Methods 8 to 21 days after birth, the Wistar rats of the experimental group were injected with daily recta glacial acetic acid 0.6% 0.3~0.5ml, the control group were injected the same dose of saline. The visceral sensitivity of 8-week-old rats was evaluated by colorectal distention，and recorded the water injection rate when the score of abdominal withdrawal reflex (AWR) was 3.The distal descending colon was taken to detect for c-kit immunohistochemisty and Western blot.Result Compared with the control group, the water injection rate of the experimental group showed a significant decrease when the score of AWR was 3(P＜0.05);C-kit masculine cells counting in colon in the experimental group significantly increased(P＜0.05);The expression of c-kit protein in colon in the experimental group significantly increased(P＜0.01).Conclusion The increase of ICC of rat with visceral hypersensitivity may be a possible cause of visceral hypersensitivity.

The influence on glycemia control for type 2 diabetes patients underwent the bariatric surgery passively

2012, 32(5):  570-573. 

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Objective: The patients with T2DM that passively underwent gastrectomy were chosen and the differences of their basic clinical data before and after the gastrectomy were analyzed to find the contribution of the factors in glycemia control. Thus the information of bariatric surgery in treating T2DM was offered in our country. Methods: The clinical data of 45 patients with T2DM that underwent gastrectomy in Peking Union Medical College Hospital from January 2001 to December 2010 have been retrospectively analyzed in this study. The patients were divided into the improved group (29 cases) and the ineffective group (16 cases) with the standard that whether the glucose level or amount of antidiabetic drug reduced for 20% after surgery or not. The data of age, body mass index(BMI), course of T2DM,preoperative blood pressure were analyzed to determine the influencing factors of the differences between the improved group and the ineffective group. Results: There was a significant difference in the course of T2DM between the improved group(5.51±1.70)and ineffective group(10.70±2.31).In the improved group and ineffective group, the average age was 61.43±11.33 and 67.09±9.10, BMI was 23.19±2.28 and 25.24±3.40 respectively. The differences were both not statistically significant. In addition, the blood pressure didn’t have significant change after the surgery in patients with both T2DM and hypertension. Conclusion: The surgical treatment of T2DM is suitable for patients with shorter courses of T2DM(less than 5 years). BMI is meaningless in choosing the patients. The sample size of this study was small and the gastrectomy was passive in the patients thus prospective studies with large samples are needed to verify this conclusion.

Causes analysis of postoperative nonunion of femoral shaft fracture treated with intramedullary interlocking nail

2012, 32(5):  574-576. 

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Objective To investigate the causes of postoperative complications of femoral shaft fracture treated with intramedullary interlocking nails Methods A total of 34 patients with femoral shaft fractures admitted to our hospital from 2009 to 2010 were retrospectively studied. They were all treated with intramedullary interlocking nails. The locking nails were removed from 6 patients with delayed union and 3 patients with nonunion to dynamise the fracture. For 1 patient with bone nonunion, the intramedullary interlocking nails were removed, and a larger one and bone grafts were applied. Results Followed up for more than 10 months, the fractures of 6 patients with delayed union and 4 patients with nonunion were completely healed. Conclusions The femoral shaft fracture healing treated with intramedullary interlocking nails depend mainly on intramembranous ossification so that periosteum invasion should be avoided during operations. Dynamisation of the fracture in time can reduce bone nonunion through endochondral ossification.

Telmisartan delay the development of diabetic nephropathy in rats

2012, 32(5):  577-578. 

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Metoprolol combined with atorvastatin ameliorated left ventricular remodeling in renovascular hypertensive rats

2012, 32(5):  581-582. 

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Research advances of microRNAs related to laryngeal carcinoma

2012, 32(5):  583-586. 

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Laryngeal carcinoma is one of the most common cancers in otolaryngology-head and neck surgery. However, its pathogenesis is unclear. Studies have shown that microRNAs (miRNAs) play an important role in the carcinogenesis of laryngeal carcinoma. The abnormal expressions of miRNAs (such as upexpression of miR-106b-25 and miR-21 etc., downexpression of miR-375 and let-7a etc.) are related with the occurrence of laryngeal carcinoma. The abnormal expressed miRNAs in laryngeal carcinoma tissues can be used as tumor markers in the diagnosis of laryngeal carcinoma. Furthermore, some miRNAs may be used as targets for the treatment of laryngeal carcinoma.

Application of clinical and interactive teaching in pathophysiology teaching

Rui FENG

2012, 32(5):  587-589. 

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Objective To training better on the excellent medical talents who can apply their knowledge of basic medical science to work, pathophysiology teaching should improve teaching methods, combine closely with clinic and enhance students’ learninginterest and the quality of education. Methods In pathophysiology teaching our teachers take the advantages of teaching and use various teaching methods, such as problem-based teaching, clinical case analysis, multimedia teaching aids combined and so on. Results In the teaching process of pathophysiology basic medical theory combines with clinical practice closely, and it is more beneficial for the students to master basic medical theory, and to use their knowledge to find and solve the problem in clinlc.Conclusion The clinical and interactive teaching is suitable to apply in the teaching of phathophysiology.

Exploration on teaching reform of medical pharmacology

Cai-ying YE

2012, 32(5):  590-592. 

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In this paper, the problems in medical pharmacology course are discussed around teaching methods, teaching material selection, student character and assessment system, and then some views are proposed, which might be beneficial for pharmacology curriculum reform in the future.