Objective To investigate the effects of miR-135a on HOXA10 expression,proliferation and apoptosis of SKOV3 cells.Methods (1) Through computer-aided algorithms,the predicted target gene of miR-135a (HOXA10)were determined.(2) miR-135a mimics,miR-135a inhibitor and negative control were transfected into SKOV3 cells,respectively.Reverse transcription (RT)-PCR,western blot analysis were used to examine the expression levels of HOXA10 at different times (24,48 and 72 hours).(3) A luciferase reporter assay was used to confirm the direct regulation between miR-135a and HOXA10.(4) SKOV3 cells proliferation at different times (24,48 and 72 hours) was detected by methyl thiazolyl tetrazolium(MTT) assay [quantified by absorbance(A)].Western blot was used to examine the expression of apoptosis-associated protein bcl-2,bax and caspase-3 in SKOV3 cells after 48 hours transfection.Results (1) HOXA10 was predicted to be the target gene of miR-135a by computer-aided algorithms.(2) RT-PCR shown that HOXA10 mRNA levels were decreased over time (24,48 and 72 hours) after miR-135a mimics transfectionin SKOV3 cells (0.94 ±0.04 vs 0.78 ±0.03 vs 0.70 ±0.03,P ＜0.05).While,the expression of HOXA10 mRNA was increased over time after miR-135a inhibitor transfection (1.14 ± 0.05 vs 1.16 ±0.03 vs 2.60 ±0.08,P ＜0.05).After transfected with miR-135a mimics or miR-135a inhibitor over 48 and 72 hours,the HOXA10 expression levels in SKOV3 cells were significantly lower or higher than each control group,respectively (all P ＜ 0.01).Western blot analysis of HOXA10 expression in SKOV3 cells confirmed the results of RT-PCR detected.(3) After cotransfection of miR-135a plasmid and pMIR-REPORT luciferase plasmid containing HOXA10,luciferase reporter assays showed that the luciferase activity reduced by 67.8％ (P ＜0.01).(4) MTT showed that SKOV3 cells growth after miR-135a mimics transfection for 48 and 72 hours were significantly lower than those in control group (0.38 ± 0.03 vs 0.52 ± 0.05,0.67 ±0.05 vs 0.75 ± 0.06 ; respectively,all P ＜ 0.05).While,SKOV3 cells transfected with miR-135a inhibitor for 72 hours grew significantly faster than that in control group (0.95 ± 0.05 vs 0.75 ± 0.06,P ＜ 0.01).After miR-135a mimics transfection,the level of bcl-2 protein was significantly lower than that in control group (0.28 ±0.06 vs 0.76 ±0.09,P ＜0.01).The activity of caspase-3 was significantly higher than that in control group (115.0 ± 2.4 vs 95.4 ± 2.1,P ＜ 0.01).While,there was no statistical difference of bax expression (P =0.142).However,after miR-135a inhibitor transfection,the expression level of bcl-2 protein was significantly higher than that in control group (0.92 ± 0.03 vs 0.76 ± 0.09,P =0.037) and the activity of caspase-3 was significantly lower than that in control group (59.5 ± 4.1 vs 95.4 ± 2.1,P ＜ 0.01).There was also no statistical difference of bax expression (P =0.066).Conclusion miR-135a may play an important role in cell proliferation and apoptosis of ovarian cancer cells by regulating HOXA10 and its downstream pathways.

Objective To retrospect the results and probe into the clinical applied vallue of the pedicled bone flap with deep iliac circumflex artery to the treatment of the femoral neck fracture and femoral neck fracture Methods Eight cases of femoral neck fracture and 21 cases nonunion treated by bone and periost flaps with deep iliac circumflex vessel35 casses treated by closure reduction and fixed with 3 trianglar nails,34 cases by arthroplastyThe results of 3 kinds of methods were contrasted and anslysed Results Twenty-nine cases treated by iliac bone and periost flaps with deep iliac circumflex artery were fellowed from 3 to 7 years,the fracture healed 35 months averagely. One case was avascular necrosis of femoral head after 9 month after operation,others received good results35 cases were treated by 3 nails fixtion healed 74 months averagely and 6 case had complications 37 casese treated by arthroplasty had complications in 8 patients,4 cases need walking-stich for walk. Conclusion Femoral neck fracture were treated by transplantation of the pedicled bone and periost flaps with deep iliac circumflex vessel is keeping with the pathological and anatomical characteristicsThe function of the hip joint is not weaken as the time goes on when the fracture is healedIt is the best methods to treat the femoral neck fracture now,the arthroplasty has many complication,so the indication must be confined

Objective To explore the Diagnostic value of real-time three-dimensional ultrasound for urethral vaginal fistulas.Methods 5 patients with urethral vaginal fistulas were checked using real-time three-dimensional ul-trasound ,respectively by the abdomen and the sonographic appearance of pudendal observation of female urethral vagi -nal interval sagittal and transverse ,sagittal and transverse section by measuring the size of the fistula ,and voiding trial or colored liquid test injection from urethral to confirm .Results 5 cases of urethra,vagina ultrasonography can dis-play and urethral vaginal interval throughout the sagittal plane and different levels of cross section ,urethral vaginal in-terval by low echo-band-low echo constitute the sandwichsign in the fistula showed partial loss ,local band echo in-terruption,urethral and vaginal local fusion ,voiding trial or from the urethra into the colored liquid test further con-firmed the fistula,fistula and verify the existence of fistula after the operation the size .Conclusion Real time three-dimensional echocardiography by urethral vaginal interval in the axial and sagittal plane ultrasound at different levels , combining with the history and voiding trial or injected with colored liquid test from the urethra ,can provide a new i-dea for diagnosis of urethral vaginal fistula .

Adult ; Female ; Humans ; Lead ; Male ; Middle Aged ; Occupational Exposure ; T-Lymphocyte Subsets ; immunology ; Young Adult

Objective To study the effectiveness of comprehensive control measures based on systematic ecological system construction to interrupt the transmission of schistosomiasis in hilly endemic regions in Sichuan Province ,so as to provide the ev?idence for adjustment of schistosomiasis prevention and control strategies. Methods A high endemic area of schistosomiasis, Panao Township of Dongpo District in Meishan City,was selected as a demonstration area. The comprehensive measures for schistosomiasis control with focus on systematic ecological management were implemented,and the income of residents,index?es of schistosomiasis control effect and so on were investigated before and after the intervention and the results were compared. Results The project based on systematic ecological system construction started in 2009 and 317.351 million Yuan was put into the construction. The construction included economic forest plant base(1 866.68 hm2,72.66%of the total farmland areas),eco?logical protection gardens(585.35 hm2)and so on. Totally 97.04% of historical areas with Oncomelania hupensis snails were comprehensively improved. In 2015,the peasants' pure income per capita increased 4 938 Yuan,with the average annual growth rate of 14.69%. All the farm cattle were replaced by the machine. The benefit rate of water improvement was increased by 52.84%and the coverage rate of harmless toilets increased by 18.30%. The positive rate of serological tests for schistosomiasis decreased from 7.69%to 3.50%,and the positive rate of parasitological tests decreased from 1.18%to 0. The area with snails was decreased from 23.33 hm2 to 0. The awareness rate of schistosomiasis control knowledge and correct behavior rate of the resi? dents increased from 85.50%and 82.60%to 95.70%and 93.90%respectively. Conclusions The comprehensive schistosomia?sis control measures based on systematic ecological management are conform to the currently actual schistosomiasis prevention and control work in hilly endemic regions,and have good ecological economic benefit and schistosomiasis control effectiveness, which provide an effectively new model of prevention and control for advancing process,consolidating the effect,finally realiz?ing goal of interruption and elimination of schistosomiasis in hilly endemic regions.

0.05). However, a significant higher percentage of CD3+CD25+ T cells was found in stable liver transplantation group as compared to healthy group ( P0.05). There was not found no regular change of serum cytokine s (IL-6, TN F-?) and adhesion molecules (ICAM-1, P-selectin) in 6 liver transplanted patien ts during post-operation from day 1 to day 30, indicating that was associated wi th the different status of patients before or after transplantation. CONCLUSIONS: Our data suggesting that increased levels of ICAM-1 and P-selectin, appears to participate in the processing of immunoregulation to transplanted livers, whereas elevated concentrations of IL-6 appear to be involv ed in the repair of the injury induced by TNF-? in allo-transplanted livers.

Objective To study the relationship between cognitive dysfunction and location of lesion and prognosis in patients with cerebral infarction.Methods 120 patients with cerebral infarction in the Second People's Hospital of Yueqing were selected.According the results of the cerebral CT or MRI,the patients were divided into primary group(58 cases)and recurrence group (62 cases).3 -10days after admission,the cognitive impairment was evaluated with the method of simple mental state examination (MMSE)scale.The stroke score of the National Institutes of health(NIHSS)in the United States and ability of daily life activity Barthel index score before the admission and after the discharge were conducted on the patients.And the relationship between different lesion location of primary or recurrent infarction,cognitive disability and prognosis were observed.Results The incidence of cognitive impairment in the ecurrence group was 90.32%,which was obviously higher than 72.41% in the primary group(χ2 =6.419,P =0.011);In the two groups,the proportion of thalamic cortex was the highest,but there was no significant difference between the two groups (χ2 =0.000,P =1.000).The NIHSS and Barthel scores of patients with thalamic infarction were (11.4 ±1.6)points and (27.5 ±3.7)points respectively,both of which were the highest(FNIHSS =47.22, PNIHSS =0.000;FBarthel index =17.77,PBarthel index =0.000);the NIHSS and Barthel indexes in each part of the two groups at discharge were lower than those before the admission (all P <0.01),and the differences at admission was not statistically significant(P >0.05);the NIHSS scores of the primary group and the recurrent group at discharge were (6.3 ±0.5)points and (7.8 ±0.7)points respectively;the Barthel scores were (24.3 ±7.3)points and (30.8 ± 6.8)points respectively.And all of them were lower than those at admission and the improvement degree of the primary group were more significant than those of the recurrence group(tNIHSS =13.43,PNIHSS =0.000;tBarthel index =5.05, PBarthel index =0.000).Conclusion Cognitive impairment after cerebral infarction is associated with lesion,and thalamic infarction is the most prone to perform cognitive disfunction with poor prognosis,and the frontal and temporal lobe are also common parts where targeted prevention is needed.

Objective To observe the protective effect of spironolactone on heart of hypertensive SD rats.Methods 30 male SD rats were randomly divided into the control group,hypertensive group(HS group),low spironolactone dose group(HSL group)and high spironolactone dose group(HSH group).The animals except the control group were treated with L-NAME 50 mg/kg/d.Then the animals of the HS group were fed with 1% sodium chloride water solution.Those of the HSL and HSH groups were treated same but the former added with spironolactone 20 mg/kg/d,the later added with spironolactone 100 mg/kg/d.8 weeks late,the animals were killed and hearts were taken out to observe following items:myocardial cross-sectional area,ratio of per-arteriolar fibrous area/arteriolar luminal area of arterioles,ratio of collagen area/total myocardial area,myocardial necrosis and inflammation.Results The myocardial cross-sectional areas in the HS group were significantly more than those in other 3 groups(P<0.01).Collagen ratios of the HS group were significantly higher than others(P<0.01).The ratios of arteriolar wall area versus luminal area in the HS group were significantly higher than those of control and HSH groups(P<0.01).Myocardial injuries in the HS group were more severe than those of other 3 groups.There were no differences between HSL and HSH groups.Conclusion Spironolactone has protective effect on heart of hypertensive SD rats.

To investigate the biological role of snake venom cystatin(sv-cystatin) in tumor progression, the cDNA of sv-cystatin amplified by PCR from pUC18-cystatin plasmid was cloned into methanol-inducible expression vector pPICZ?A. The linearized recombinant plasmid pPICZ?A-cystatin was transfered into Pichia pastoris, strain GS115 by electrophoration. Transfermants with phenotype Mut+ selected were identified by PCR analysis and induced in 1.0% methanol. The reombinant sv-cystatin protein was examined by SDS-PAGE, Western blot analysis. The molecular mass of expression product was about 14 kDa and approximately 16 mg/L of recombinant sv-cystatin was produced from one of GS115-cystatin transformants. The chromatography purified protein could reduce the activity of papain. The ability of B16F1 cells treated with recombinant sv-cystatin to invade the reconstituted basement membrane decreased significantly (P

Background The pathological basis of human primary open angle glaucoma mainly is the degeneration of trabecular meshwork and Schlemm canal.As the outflow pathway of aqueous humor, the tissue origin and characteristics of trabecular meshwork remain less clear.Studying the characteristics of trabecular meshwork may contribute a new thought to the treatment of primary open angle glaucoma.Objective This study was to explore the histological property of trabecular meshwork by detecting the expression of D2-40, a lymphatic biomarker,and CD31,CD34 and smooth muscle actin (SMA), some vascular endothelial biomarkers.Methods Twenty specimens of adult eyeballs were collected from Zhongshan Ophthalmic Center of Sun Yat-sen University,including l0 eyeballs from accident death donors,3 enucleated eyeballs due to posterior segment of choroidal malignant melanoma and 7 orbital exenterated eyeballs.The series ocular meridian sections with the thickness of 4 μm were prepared for the haematoxylin-eosin staining.Immunohistochemistry was employed to detect the expression of D2-40,CD31 ,CD34 and SMA in trabecular meshwork and Schlemm canal.Results Intact structure of chamber angle was observed under the optical microscope by haematoxylin-eosin staining.Human trabecular meshwork tissue was presented with meshlike structure,while Schlemm canal showed the lumen structure.No abnormal substance and periphery synechia were found.D2-40 was strongly expressed in the trabecular meshwork endothelial cells with staining score for 3 points, but CD31, CD34 and SMA were negatively expressed.In contrast, CD31, CD34 and SMA rather than D2-40 were positively expressed in the Schlemm canal.In addition, CD31 and CD34 were also positively expressed in the vascular endothelial cells around the trabecular meshwork.Conclusions The trabecular meshwork expresses lymphatic biomarker rather than vascular biomarkers.This result indicates that trabecular meshwork of human eyes is lymphatic vessel-like tissue.