Ojective To investigate therapeutic results of conservative operation with laparoscopy for all kinds of patients with tube pregnancy. Methods 62 patients with tube pregnancy were performed the conservative surgery, fenestration on the position of tube pregnancy or take out embryos from the tube crevice combined with injection of MTX under laparoscopy. Results all of 61 patients' tube were retained successfully. no complications occurred, urine HCG recovered to normal levels two weeks after operation. Only one patient continue to pregnancy and completely recovered after intramuscular injection MTX. 18 patients were carried out HSG 1～4 months after operations. 15 patients' tube among them were unobstructed. Opening rate was 88.3%. Within postopeative four years, 12 patients were repregnant. Pregnancy rate was 66.7%. Conclusions Tube pregnancy conservative treatments under laparoscopy not only is safe and minimal invasion, but also had less complications, therefore it is a more suitable method for retaining the fertile females' tubes.

Objective To investigate the significance of optimization of bedside Gram staining of sputum smear in the early diagnosis and antimicrobial treatment for ventilator-associated pneumonia(VAP)patients. Methods The data of patients with VAP undergoing mechanical ventilation over 48 hours in the Department of Critical Care Medicine of Tianjin Fourth Central Hospital from June 2009 to June 2014 were analyzed. The patients were divided into two groups according to whether or not bedside Gram staining of sputum smear was used or not. The sputum samples from lower respiratory tract of all VAP patients were collected daily with tracheal catheter. In empirical examination group(from June 2009 to December 2011,n=43),the patients received antibiotics at the time of onset of VAP, selection of antibiotics depended on the information of bacterial epidemiology of the intensive care unit(ICU),and also existence of high risk factors of multi-drug resistant bacteria. In target treatment group(from January 2012 to June 2014,n=43),the patients received antibiotics according to the results of bedside instant sputum smear examination and empirical antibiotic regime. The correlation between the results of sputum smear examination and culture result was analyzed. The levels of body temperature,white blood cell(WBC)count,procalcitonin(PCT)level,and high sensitivity C-reactive protein(hs-CRP)were measured on the 1st day and 3rd day. The length of antibiotics treatment, duration of mechanical ventilation,and the time of ICU stay were recorded for both groups. Results There were 512 qualified sputum specimens for culture,from which 336 pathogens were found,and 358 strains of pathogenic bacteria were found from microscopic examination of 512 qualified sputum smear. The coincidence rate of results of bedside examination of sputum smear and that of sputum culture was 78.32%(401/512). The diagnostic acumen of the former was 85.42%(287/336),specificity was 64.77%(114/176),positive predictive value was 80.17%(287/358),and negative predictive value was 74.03%(114/154). On the 1st day,no statistical differences in infection index between the two groups could be found,but on the 3rd day,the results were significantly improved in both groups. Compared with the empirical treatment group,the body temperature,WBC,PCT and hs-CRP in the target treatment group were significantly lower〔body temperature(℃):36.83±0.69 vs. 37.64±0.71,WBC(×109/L):7.91±2.75 vs. 9.66±3.39,PCT(μg/L):7.14±3.89 vs. 10.14±4.32,hs-CRP(mg/L):12.24±6.28 vs. 15.54±5.94,P<0.05 or P<0.01〕. Compared with the empirical treatment group,the time of antibiotics use(days:6.00±2.55 vs. 9.20±3.46), the duration of mechanical ventilation(days:5.00±1.73 vs. 7.00±1.94),and the length of ICU stay(days:7.43±1.72 vs. 12.57±4.16)were significantly shortened(P<0.05 or P<0.01). Conclusions The results of bedside sputum examination and sputum culture showed a good correlation,and the former is helpful in early diagnosis and treatment of VAP. The result of high quality sputum smear in significant in guiding the first choice of antibiotics,reduce the time of antibiotic use,shorten the duration of mechanical ventilation and the length of ICU stay,and improve the outcome of the patients.

Objective To study the association between three single nucleotide polymorphisms (SNP; rs2241766,rs1501299 and rs12495941) variants of the adiponectin gene (ADIPOQ) and polycystic ovary syndrome (PCOS) in PCOS family trios.Methods A total of 224 unrelated PCOS probands,their biological parents were recruited.Anthropometric variables such as waist circumference (WC),hip circumference (HC),height and weight were measured in all subjects during the first visit to the outpatient department.Body mass index (BMI) and waist-hip ratio (WHR) were calculated.Serum fasting glucose (FBG),fasting insulin (FINS),total cholesterol (TC),triglycerides (TG),low-density lipoprotein and highdensity lipoprotein cholesterol (HDL) levels were measured.PCOS patients were divided into two groups based on BMh group A (BMI＜25 kg/m2) and group B (BMI≥25 kg/m2).Parents of PCOS patients were accordingly categorized into group C,D (fathers) and group E,F (mothers).The transmission disequilibrium test (TDT) was used to analyze the association between three SNP of ADIPOQ and PCOS.Results (1) A significant positive association was detected between SNP rs1501299 and PCOS (x2=7.093,P=0.008).However we failed to find significant overtransmission of the other two SNP rs2241766 and rs12495941 from parents to PCOS offsprings (x2=1.620,P=0.203; x2=0.713,P=0.398).(2) Linkage disequilibrium (LD) was analyzed in the subjects,rs1501299 and rs2241766 were in weak LD (r2=0.063,D'=0.621).(3) The levels of WC,HC,WHR,testosterone,TG,HDL and FINS were significantly differences between obese and lean PCOS patients (P＜0.05).While in fathers we only found WC,HC,TC levels being statistically different (P＜ 0.05).Mothers of obese PCOS patients had increased levels of FINS compared with mothers of lean PCOS patients (P＜0.05).The genotype frequencies of the three SNP were not different in obese and lean PCOS patients and their parents (P＞0.05).Conclusions TDT confirms that SNP rs1501299 in the ADIPOQ is significantly associated with the risk of PCOS in the Chinese Han population.The three SNP of the ADIPOQ were not associated with the obesity of PCOS.

Objective To comprae the difference of Sex hormone,serum and follicular fluid anti Mullerian hormone (AMH) level tube,number of retrieved egg,the number of cleavage rate,fertilization oocytes,the number of available embryos and total embryo on in vitro fertilization and embryo transfer (IVF-ET) of different reaction groups,to explore the relationship between AMH and pregnancy outcome of IVF-EH.Methods Eighty-one cases of IVF-ET assisted pregnancy were divided into low ovarian response group,normal ovarian response group and high ovarian response group according to the response of the super ovulation.Serum sex hormone and AMH of all patients were detected on the third day of menstruation,on the day of HCG injection,oocyte pick-up (OPU) and embryo transfer (ET),AMH in FF were detected too.The difference of oocytes,cleavage rate,embryo quality and the indicators mentioned above among the three groups were compared.All of the above indicators were compared between pregnancy group and unpregnancy group.Results (1) The level of AMH on the third day of menstruation ((1.76 ± 0.47) μg/L,(3.45 ± 1.01) μg/L,(6.34 ± 1.29) μg/L,F =2.435,P＜0.001),HCG injection ((1.09±0.13)μg/L,(1.29±0.29)μg/L,(2.64±0.51)μg/L,F=1.542,P=0.001) and embryo planting((1.32±0.33) μg/L,(1.62±0.39) μg/L,(1o 91±0.41) μg/L,F =1.573,P ＜0.001) all were statistically significant in three groups.On day of OPU,the levels of serum AMH ((0.95±0.21)μg/L,(1.15±0.29) μg/L,(1.74±0.41) μg/L,F =12.573,P＜0.001) and AMH in FF((5.82±1.19) μg/L,(6.92±1.05) μg/L,(7.79±1.39) μg/L,F =9.83,P＜0.001) were statistically differences in the three groups.(2)The number of oocytes were positively correlated with AFC,based AMH,levels of E2,P and AMH on the day of HCG injection and OPU,E2 and AMH on the day of ET and AMH in FF(P＜0.05).(3)AMH in FF in pregnancy group was (6.22±0.82)μ g/L,significantly higher than those without pregnancy group ((5.31 ±0.71)μg/L,P =0.037).Conclusions Levels of AMH in serum and follicular fluid are good predictors of ovarian response in IVF-ET.The level of serum AMH can only predict ovarian response,but can not effectively predict pregnancy outcome.The level of AMH in FF is a good predictor of pregnancy outcome.

Objective To assess the effect of a noninvasive equipment (the human biological stimulation feedback instrument) for the screening of metabolic syndrome (MS). Methods The samples were selected from the International Health Care Center of the Second Affiliated Hospital of Zhejiang University College of Medicine. From January to December 2012, according to the inclusion criteria, a total of 5674 consecutive participants (3 437 men, 2 237 women) without medical history of diabetes mellitus underwent general health screening in this hospital, which included questionnaire investigation, all MS components and the risk assessment of the human biological stimulation feedback instrument. MS was defined according to 2004 the Chinese Diabetes Society (CDS) criteria. Results The 5 674 subjects were divided into three groups in accordance with the human biological stimulation feedback instrument risk score: the normal group, metabolic abnormalities low risk group, metabolic abnormalities high risk group. The detection rate of MS corresponding to the three groups was 8.73% (251/2 876), 29.32% (612/2 087), 32.77% (233/711), respectively (trend χ2=372.14,P<0.001). With the increase of risk score, the detection rate of MS increased. A total of 1 096 (male 879, female 217) were diagnosed MS by CDS criteria. The area under the curve (AUC) of the test was 0.674 (95%CI 0.658-0.691, P<0.000 1)for predicting MS. The corresponding cutoff value was 25.00%, the sensitivity, specificity, positive predictive value and negative predictive value of diagnosis of MS was 77.80%, 56.00%, 30.20%and 91.30%, respectively;409 subjects who had no history of MS and diabetes mellitus follow up for two years, the AUC of the test was 0.719 (95%CI 0.644-0.794)for predicting MS. The corresponding cutoff value was 25.00%, too. And the sensitivity and specificity of diagnosis of MS was 78.00%and 59.80%, respectively. The detection rate of MS was positively correlated with the risk score. Conclusion We found that the human biological stimulation feedback instrument is a potential screening approach for early predicting MS in health checkup with simple and noninvasive procedure and high negative predictive value, although its diagnostic efficiency is not so good. And it might have a warning effect on the progress of MS.

Purpose To study the effect of China cobra venom active factor(CCVAF) from China cobra venom on endothelial cells and its mechanism.Methods MTT experiment was adopted to evaluate the effect of CCVAF on bovine arteria pulmonalis vascular endothelial cells(BAVEC).The Eosin-Coomassie brillient blue and rhodamine-phalloidin method was used for actin cytoskeleton.Flow cytometry for [Ca~(2+)]_i and spectrophotometry were used for lactate dehydrogenase(LDH) and nitrogen oxide(NO) levels in cell culture supernatant.Results CCVAF(0.625-20 μg/mL) inhibited the proliferation of BAVEC in dose-dependent manner,and IC50 of CCVAF on BAVEC was 2.45 μg/mL. After CCVAF and BAVEC coincubation, it was showed that regression of intercellular conjunctions and disorder of F-actin distribution occurred. The content of [Ca~(2+)]_i, [LDH] and [NO] increased respectively.Conclusion CCVAF can inhibit BAVEC proliferation and it maybe associated with the change of cytoskeleton and increasing of [Ca~(2+)]_i,[LDH] aod [NO].

Objective To provide objective evidences for early screening of the hypothalamic-pituitary-adrenal(HPA)axis function in diabetic patients, the relationship between HPA axis function and diabetes mellitus was systematically reviewed. Methods PubMed, Ovid, Embase, Web of science Databases were retrieved from articles reporting the relationship between HPA axis function and diabetes mellitus. Meta-analysis were carried out by Stata 12. 0 software for heterogeneity test and data merged among articles for the inclusion and exclusion criteria. Results Our meta-analysis showed the following results: Compared with control group, the basal plasma cortisol level in total diabetes group, T1DM group, T2DM group, diabetes patients(with or without chronic complications)combined with hypertension was significantly higher. Compared with control group, postdexamethasone cortisol level was markedly higher in diabetes group. There was no significant difference in 24 h urinary free cortisol(24h UFC)and ACTH levels between diabetes group and control group. Our qualitative evaluation shows the following results: the differences of awaking salivary cortisol and cortisol awakening response(CAR)between the two groups were inconsistent among all included studies. Most studies showed that CAR was lower in diabetic patients. Conclusion The present analysis has demonstrated that HPA axis dysfunction exists along with diabetes mellitus, and diabetic comorbidity may further worsen the HPA axis dysfunction.

Background Xenotransplantation is arousing more attention of researchers because of the lack of corneal donors.Biological corneal scaffolds constructed by porcine corneal acellular stroma appears to have an acceptable biocompatibility.However,its clinical effects and the histomorphological features in the corneal tissue of receipts' are still unclear.Objective This study was to evaluate the viability of bioengineered corneas as a new material of human lamellar keratoplasty and observe the in viva biological features after human keratoplasty under the laser confocal microscope.Methods A prospective serial cases observational study was carried out.Fifteen eyes of 15 patients with infectious keratitis were enrolled in Henan Eye Hospital from February to August 2014 under the approval of Ethic Committee of Henan Eye Hospital and informed consent of each patient, including 4 eyes of fungal keratitis,1 eye of bacterial keratitis, 9 eyes of mixed infectious keratitis and 1 eye of leucoma.Corneal lamellar transplantation was performed on the eyes with the bioengineered corneas as grafts and the follow-up time was one year.The survival of grafts was assessed and scored, and the indices including corneal transparency,neovascularization, graft lysis or inflammatory recurrence and visual acuity (logMAR) were evaluated.The morphology and density of corneal epithelial cells and endothelial cells, corneal stroma and subepithelial neural fibers were examined by laser confocal microscope 3,6,9 and 12 months after surgery.Results Postoperative inflammatory response was seen 3 days and disappeared 7 days after surgery.The grafts were clear 1 month after surgery, and no corneal dissolution was found during the follow-up duration.Glaucoma occurred in 1 eye at 6 months and graft rejection occurred in another eye 12 months after surgery.The logMAR, corneal transparency scores and corneal neovascularization scores improved after surgery in comparison with before surgery,with a significant difference among various time points (x2 =92.63,59.37,10.50 ,all at P＜0.05).Complete epithelization of grafts was seen.Compared with the contralateral eyes,the morphology of epithelial cells was similar and endothelial cells were enlarged in the operated eyes.In addition,no stromal cell structure was seen in the corneal stroma in the operative eyes.Subepithelial nerve fibers appeared in 8 eyes at 6 months, but the fiber density was lower in the operated eyes than that in the contralateral eyes.Significant difference was found in epithelial cell density among different time points (F=1.48, P =0.22).The endothelial cell densities were (2 542 ± 119), (1 895 ± 129), (1 869 ± 135), (1 854 ± 101) and (1 844 ± 103)/mm2 before surgery and 3, 6, 9 and 12 months after surgery, showing significant differences between preoperation and postoperative time points (all at P＜0.05).The subepithelial nerve fibers densities were (1.26± 0.13),(3.62±0.81) and (5.98±0.44) mm/mm2 at 6,9 and 12 months after surgery,with significant differences between adjacent two time points (t'=-8.16 ,-7.24;both at P=0.00).Conclusions Bioengineered corneal grafts survive well in human eye after lamellar transplantation,which can reconstruct ocular surface and improve the visual acuity.Biological cornea can replace human corneal materials.

Objective To investigate the effects of high-fat diets and rosiglitazone treatment on the expressions of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) and mitofusin-2 (Mfn2) in skeletal muscle of aged rats. Methods Male wistar rats aged 21-23 months were randomly divided into old control group (OC; n=20), high-fat diets group (HF; n=20) and high-fat diets plus rosiglitazone treatment group (RSG; n=20). Male wistar rats aged 4-5 months were selected as young control group (YC; n=20). Insulin sensitivity was evaluated by hyperinsulinemic-euglycemic clamp technique at the end of the 4th and 8th week. The expressions at mRNA and protein levels of PGC-1α and Mfn2 in skeletal muscle were determined by polymerase chain reaction and Western blotting technique. Results (1)After 8 weeks, the levels of free fatty acid [(0. 68±0. 18)mmol/L, (0.82±0. 23) mmol/L and (1. 53±0.40) mmol/L], triglyceride [(0.53±0.13) mmol/L,(0. 63±0. 17) mmol/L and (1.08±0.30) mmol/L]and muscle triglyceride [(1.09±0.17) mmol/L,(1.34±0. 20) mmol/L and (2.07±0. 30) mmol/L]in YC group, OC group and HF group were significantly increased and glucose infusion rate [(30.4±4. 2) mg·kg~(-1)·min~(-1), (20.9±2.2) mg·kg~(-1)·min~(-1) and (12. 0±1.9) mg·kg~(-1)·min~(-1)]was significantly decreased as compared with pre-high fat diet, respectively. The levels of fasting free fatty acid [(0.93±0.29) mmol/L], triglyceride [(0.62±0.12) mmol/L]and music triglyceride [(1.68±0.28) mmol/L]in RSG group were significantly decreased and glucose infusion rate [(16.7±1.7) mg·kg~(-1)·min~(-1)]was significantly higher than in HF group. (2)In skeletal muscle, the expressions at mRNA and protein levels of PGC-1α and Mfn2 decreased in OC group compared with YC group (all P<0.01). The expressions of PGC-1α and Mfn2 were lower significantly in HF group than in OC group, and were higher significantly in RSG group than in HF group (all P<0.01). Conclusions The aged rats fed the high-fat diets develop insulin resistance with decreased expressions of PGC-1α and Mfn2 in skeletal muscle. Insulin sensitivity is improved with rosiglitazone treatment by increasing expressions of PGC-1α and Mfn2 in skeletal muscle.

Aim To evaluate the mechanism of apopto-sis induced by the isoliquiritigenin in A375 human ma-lignant melanoma cells. Methods Sulforhodamine B ( SRB) method was used to determine the A375 cell viability;acridine orange/ethidium bromide ( AO/EB) and Hoechst 33258 staining were used to observe the morphological changes of apoptotic cells; flow cytome-try was used to detect A375 cell apoptotic rate;DCFH-DA was applied to determine the changes of total intra-cellular ROS in A375 cells;JC-1 method was used to measure the changes of mitochondrial membrane poten-tial;the kits methods were used to determine the con-tent of ATP, lactic acid and glucose in A375 cell which was treated with different concentrations of isoliquiritigenin. Results Isoliquiritigenin could in-hibit A375 cell proliferation in a concentration-depend-ent manner; A375 cells showed obvious apoptosis charateristics after treatment by isoliquiritigenin, and the apoptosis rate increased with increasing concentra-tion of isoliquiritigenin. The level of total intracellular ROS in A375 cells increased obviously after dealing with different concentrations of isoliquiritigenin;in ad-dition, the mitochondrial membrane potential, the lev-els of intracellular ATP,lactic acid and the level of glu-cose uptake all declined. Conclusions These find-ings demonstrate that isoliquiritigenin can induce apop-tosis of A375 cells. The mechanism may be related to elevation of ROS level and reduction of aerobic glycoly-sis level.