ABSTRACT:Objective To explore the effects of miRNA-1246 (miR-1246)on cell proliferation,invasion and migration in human cervical squamous cell carcinoma (CSCC)cell line SiHa.Methods SiHa cells were assigned into 3 groups:miR-1246 analog group,miR-1246 antagonist group and control group.Transfection efficiency was determined.The MTT assay,transwell assay and wound healing assay were performed respectively to evaluate the proliferation,invasion and migration abilities of SiHa cells.Western blot was carried out to detect the expression of thrombospondin-2 (THBS2)before and after transfection.A THBS2 3’-UTR-containing dual luciferase plasmid was synthesized and co-transfected with miR-1246 into SiHa cells to observe the luciferase enzyme activity.Results MTT assay,transwell assay and wound healing assay revealed that the abilities of proliferation,migration and invasion were significantly enhanced (P<0.01)in SiHa cells transfected with miR-1246 analog,but suppressed in SiHa cells transfected with miR-1246 antagonist.Western blot showed that SiHa cells transfected with miR-1246 analog had significantly decreased THBS2 expression (gray value = 6 .2 8 ± 1 0 .2 2 , P=0 .0 1 3 ) while those transfected with miR-1246 antagonist had significantly increased THBS2 expression (gray value = 12.90±19.81, P=0.037).After co-transfected with miR-1246 and THBS2 3’-UTR-containing plasmid,SiHa cells exhibited a decreased level of luciferase enzyme expression.Conclusion miR-1246 promoted the proliferation,invasion and migration of CSCC SiHa cell, and it might promote CSCC tumorigenesis and progression by suppressing the expression of its target gene THBS2 .

Objective To study the distribution status and clinical significance of human papillomavirus(HPV) infection geno‐types in condyloma acuminate(CA) tissues of vulva ,vagina and cervix .Methods The gene‐chips combined with polymerase chain reaction (PCR) technology were utilized for detecting 23 kinds of HPV genotypes in tissue specimens from 63 cases of vulval CA , 61 cases of vaginal CA and 65 cases of cervical CA .Their clinical pathological data were analyzed .Results In 63 cases of vulval CA ,56 cases were HPV positive with the HPV infection rate of 88 .89% (56/63) ,in 61 cases of vaginal CA ,55 cases were HPV positive with the HPV infection rate of 90 .16% (55/61) ,and in 65 cases of cervical CA ,62 cases were HPV positive with the HPV infection rate of 95 .39% (62/65) .Conclusion HPV infection is closely related to the CA pathgenesis in vulva ,vagina and cervix . HPV6 and HPV 11 are main stream genotypes ,in which vulval CA is most common .The gene‐chips combined with PCR technology is a method suitable for HPV typing diagnosis ,and has the characteristics of good sensitivity and high specificity ,which has an im‐portant significance for clinical diagnosis ,treatment and vaccine study of CA in femal vulva ,vagina and cervix .

To explore the causes of the postoperative complications of the penile elongation and the measures to prevent them in order to raise the success rate of the penile elongation. 1,000 patients who had received the penile elongation were reviewed and analyzed for the causes of postoperative complications, and the measures of prevention and treatment were discussed. Our results showed that, of the 1,000 cases, 64 had the postoperative complications, including 20 cases of edema of prepuce, 15 cases of flap necrosis, 12 hematoma, 9 infections, and 8 cases of fat and clumsy penis. It is concluded that correct operative manipulation, strict aseptic measures and necessary postoperative care and management could avoid or reduce the postoperative complications. When complications happened, a satisfactory result can be achieved with timely and correct treatment in the majority of the patients.
Edema/etiology ; Edema/prevention & control ; Hematoma/prevention & control ; Penis/*abnormalities ; Penis/injuries ; Penis/*surgery ; Postoperative Complications/*prevention & control ; Postoperative Complications/therapy ; Reconstructive Surgical Procedures/methods ; Surgical Flaps ; Surgical Wound Infection/etiology ; Surgical Wound Infection/prevention & control ; Urologic Surgical Procedures, Male/methods

OBJECTIVETo explore the clinicopathologic factors associated with prognosis in patients with rectal cancer.

METHODSClinicopathologic data of 2414 patients with rectal cancer, treated in the Affiliated Tumor Hospital of Harbin Medical University from May 1976 to December 2003, were analyzed retrospectively. Cox regression model was used to assess independent factors associated with prognosis.

RESULTSThe median survival time was 58 months and the 5-year overall survival rate was 45.1%. Tumors were stage I( in 75.2%, stage II( in 48.1%, stage III( in 21.3%, and stage Ⅳ( in 8.8% of the patients. The 5-year overall survival rates during the three study periods were 41.2%(1976-1986), 43.0%(1987-1996), and 49.1%(1997-2003)(P<0.01). On univariate analysis, age, time at diagnosis, histological type, distant metastasis, type of surgery, intent of surgery, gross morphology, pathologic T stage, lymphatic invasion, bowel obstruction, and TNM stage showed statistically significant association with survival. Independent prognostic factors on multivariable analysis were gross tumor morphology chi-squared value(CV):68.744, pT(CV:81.344), lymphatic invasion(CV:42.951), bowel obstruction(CV:37.856) and TNM stage(CV:85.329).

CONCLUSIONSSurvival in patients with rectal cancer is improved over time. TNM stage is the most important prognostic factor for survival in patients with rectal cancer.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Rectal Neoplasms ; pathology ; Regression Analysis ; Retrospective Studies

Objective To compare the accuracy of inferior vena cava (IVC) ultrasound,central venous pressure (CVP) and pulse indicator continuous cardiac output (PiCCO) in predicting fluid responsiveness in septic shock patients with myocardial depression.Methods Fifty patients with septic shock complicated with myocardial depression,aged ＞ 18 yr,were enrolled in the study.The method for fluid replacement test was as follows:0.9％ sodium chloride injectionor lactated Ringer's solution 500 ml was intravenously infused over 20 min,and positive fluid responsiveness was defined as an increase in PiCCO-monitored CO＞ 10％ after fluid replacement.Patients were divided into positive fluid responsiveness group and negative fluid responsiveness group according to the fluid replacement test.PiCCO was used to monitor stroke volume (SV),stroke volume variation (SVV) and pulse pressure variability (PPV) before fluid replacement,CVP was measured,and the diameter of IVC (d-IVC) and variability of IVC (v-IVC) were measured by bedside ultrasound.The receiver operating characteristic curve was drawn to evaluate the accuracy of each index in predicting fluid responsiveness.Results Compared with negative fluid responsiveness group,d-IVC was significantly decreased and v-IVC,SVV and PPV were increased before fluid replacement in positive fluid responsiveness group (P＜0.01).Three point five mmHg,14.5 mm,16.5％,17.0％ and 11.5％ were selected as the cutoff value for CVP,d-IVC,v-IVC,SVV and PPV,respectively.The sensitivity of CVP,d-IVC,v-IVC,SVV and PPV in predicting fluid responsiveness was 100％,92％,80％,44％ and 68％,respectively,and the specificity was 28％,80％,72％,100％ and 72％,respectively,and the area under the receiver operating characteristic curve (95％ confidence interval) was 0.609 (0.450-0.768),0.862 (0.750-0.974),0.777 (0.642-0.911),0.734 (0.596-0.873) and 0.733 (0.594-0.872),respectively.Conclusion PiCCO and IVC ultrasound provide higher accuracy in predicting fluid responsiveness in septic shock patients with myocardial depression,and IVC ultrasound is more superior because of non-invasive examination.

Objective To observe the expressions of oxidized low density lipoprotein ( OxLDL ) receptor CD36 in kidney tissue of diabetic rats and in tubular cells incubated with OxLDL, and to explore the association of CD36 with the tubular injury and renal fibrosis in the process of diabetic nephropathy. Methods Diabetic rat model with hyperlipidemia was established by feeding with high sugar and fat diet and injection of low dose streptozotocin intraperitoneally. The expression of CD36 in kidney tissues was analyzed immunohistochemically. Meanwhile, the tubular sclerosis and fibrosis injury index were estimated and calculated. NRK-52E cells were stimulated with 50 mg/L OxLDL for 5, 10, 24, and 48 h, or 100 and 150 mg/L OxLDLs for 2 and 3 days. The protein expression of CD36 was detected by Western blot. Results The expression of CD36 in the renal tubulointerstitium of diabetic rats was increased comparing to that in control rats, and was localized mainly at tubular region. The renal tubular damage index(STI)ofdiabetesgroupwashigherthanthatincontrolgroup(5.54±1.5vs0.65±0.15,P<0.05). OxLDL stimulated CD36 expression in NRK-52E cells in a dose-and time-dependent manner. Conclusion The expression of CD36 was increased in renal tubular of diabetic rats, in consistent with STI. OxLDL increased CD36 expression in NRK-52E cells. These results suggest that the expression of CD36 is associated with renal tubular damages in experimental rat diabetes.

Objective To compare the accuracy of the lung ultrasound and pulse indicator continuous cardiac output (PiCCO) in predicting volume responsiveness in septic shock patients with myocardial suppression.Methods Fifty patients of both sexes with septic shock complicated with myocardial depression,aged ≥ 18 yr,were enrolled in the study.The method for fluid replacement test was as follows:0.9％ sodium chloride injection or lactated Ringer's solution 500 ml was rapidly infused intravenously.PiCCO was used to monitor cardiac output,stroke volume,extravascular lung water index (EVLWI),pulmonary vascular penetration index (PVPI),intra-thoracic blood volume index and global end-diastolic volume index.The chest wall was scanned by ultrasound and the lung ultrasound score (LUS) was assessed.Positive fluid responsiveness was defined as an increase in PiCCO-monitored cardiac output＞ 10％ after fluid replacement.Patients were divided into positive fluid responsiveness group and negative fluid responsiveness group according to the results of fluid resuscitation test.The receiver operating characteristic (ROC) curve was drawn to evaluate the accuracy of each index in predicting fluid responsiveness.Results Compared with negative fluid responsiveness group,EVLWI,PVPI and LUS were significantly decreased,the number of patients in whom the bilateral anterior chest wall presented as B-line was increased in positive fluid responsiveness group (P＜0.01).The area under the ROC curve and 95％ confidence interval of LUS in predicting volume responsiveness was 0.836 (0.720-0.952),the cutoff value was 14.5 points,sensitivity 88％,and specificity 72％.The area under the ROC curve and 95％ confidence interval of EVLWI in predicting volume responsiveness was 0.818 (0.701-0.936),the cutoff value was 9.6 ml/kg,sensitivity 84％,and specificity 72％.The area under the ROC curve and 95％ confidence interval of PVPI in predicting volume responsiveness was 0.720 (0.575-0.865),the cutoff value was 1.55,sensitivity 92％,and specificity 52％.Conclusion The lung ultrasound and PiCCO produce higher accuracy in predicting volume responsiveness in septic shock patients with myocardial suppression;LUS has more advantages due to non-invasive examination.

This study is to evaluate the therapeutic effect of fibroblast growth factor 21 (FGF21) on NAFLD in MSG-IR mice and to provide mechanism insights into its therapeutic effect. The MSG-IR mice with insulin resistance were treated with high dose (0.1 micromol.kg-1d-1) and low dose (0.025 micromol.kg-1d-1) of FGF21 once a day for 5 weeks. Body weight was measured weekly. At the end of the experiment, serum lipids, insulin and aminotransferases were measured. Hepatic steatosis was observed. The expression of key genes regulating energy metabolism were detected by real-time PCR. The results showed that after 5 weeks treatment, both doses of FGF21 reduced body weight (P<0.01), corrected dyslipidemia (P<0.01), reversed steatosis and restored the liver morphology in the MSG model mice and significantly ameliorated insulin resistance. Additionally, real-time PCR showed that FGF21 significantly reduced transcription levels of fat synthetic genes, decreased fat synthesis and promoted lipolysis and energy metabolism by up-regulating key genes of lipolysis, thereby liver fat accumulation was reduced and liver function was restored to normal levels. In conclusion, FGF21 significantly reduces body weight of the MSG-IR mice, ameliorates insulin resistance, reverses hepatic steatosis. These findings provide a theoretical support for clinical application of FGF21 as a novel therapeutics for treatment of NAFLD.
Animals ; Body Weight ; drug effects ; Dose-Response Relationship, Drug ; Dyslipidemias ; metabolism ; Energy Metabolism ; drug effects ; Fatty Liver ; chemically induced ; complications ; Female ; Fibroblast Growth Factors ; administration & dosage ; pharmacology ; therapeutic use ; Insulin Resistance ; Lipolysis ; drug effects ; Liver ; metabolism ; pathology ; Male ; Mice ; Non-alcoholic Fatty Liver Disease ; drug therapy ; Sodium Glutamate

To explore the causes of the postoperative complications of the penile elongation and the measures to prevent them in order to raise the success rate of the penile elongation. 1,000 patients who had received the penile elongation were reviewed and analyzed for the causes of postoperative complications, and the measures of prevention and treatment were discussed. Our results showed that, of the 1,000 cases, 64 had the postoperative complications, including 20 cases of edema of prepuce, 15 cases of flap necrosis, 12 hematoma, 9 infections, and 8 cases of fat and clumsy penis. It is concluded that correct operative manipulation, strict aseptic measures and necessary postoperative care and management could avoid or reduce the postoperative complications. When complications happened, a satisfactory result can be achieved with timely and correct treatment in the majority of the patients.
Adolescent ; Adult ; Aged ; Edema ; etiology ; prevention & control ; Hematoma ; prevention & control ; Humans ; Male ; Middle Aged ; Penis ; abnormalities ; injuries ; surgery ; Postoperative Complications ; prevention & control ; therapy ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; Surgical Wound Infection ; etiology ; prevention & control ; Urologic Surgical Procedures, Male ; methods

OBJECTIVETo achieve high expression of human renal cell carcinoma-associated antigen G250 in Escherichia coli.

METHODSThe gene fragments encoding the protein obtained by PCR was cloned into prokaryotic expression vector pET32a(+) and expressed in E. coli Rosseta. The immunogenicity of the recombinant protein was evaluated by Western blotting.

RESULTSThe plasmid pET32a(+)/G250 was constructed and expressed in E. coli Rosseta successfully. Western blot analysis showed that the recombinant protein could be specifically recognized by monoclonal antibody M75.

CONCLUSIONEfficient G250 expression is achieved in prokaryotic expression system, which may facilitate further functional study of the protein and its monoclonal antibody preparation.

Antibodies, Monoclonal ; immunology ; Antibody Specificity ; immunology ; Antigens, Neoplasm ; genetics ; immunology ; metabolism ; Biomarkers, Tumor ; genetics ; immunology ; metabolism ; Blotting, Western ; Carbonic Anhydrase IX ; Carbonic Anhydrases ; genetics ; immunology ; metabolism ; Cloning, Molecular ; Escherichia coli ; genetics ; Gene Expression ; Humans ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism