Objective To evaluate short-term clinical effects of modified total pelvic floor reconstruction surgery in treatment of severe pelvic organ prolapse.Methods Thirty-nine cases with severe pelvic organ prolapse including vault prolapse diagnosed by pelvic organ prolapse quantification (POP-Q) system received modified total pelvic floor reconstruction surgery.Clinical parameters associated peri-operative period and 12 months after surgery and complications were analyzed Results Median operation time was 70 minutes (30-240 minutes),median blood loss was 100 ml (10-200 ml).Seventy-seven percent (30/39) patients were able to micturate spontaneously in the next morning after surgery with postvoid residual volume less than 100 ml (0-650 ml).No severe intra-operative complications were recorded and the rate of postoperative morbidity was 20% (8/39 ).Median post-operative hospital stay was 4 days (1-11 days).The patients were followed up at median 24 months(13-29 months).According to POP-Q system evaluation,the successful rate of operation reached 100% .Two cases (5%,2/39) were recorded as symptomatic recurrence which manifested as posterior wall prolapse within 24 months after operation.During follow-up,8% (3/39) patients were found to have erosion within 7 months after surgery,and urgent urinary incontinence was observed in 5% (2/39) cases,while constipation occurred in 8% (3/39) cases.The most remarkable complication was dyspareunia (36%,5/14); while 50% (7/14) experienced better sexual life after surgery.Conclusions Modified total pelvic reconstruction is a safe,efficient and micro-invasive surgery in treatment of severe pelvic organ prolapse.However,its influence on post-operative sexual life should be concerned.

Objective To screen and identify the dengue virus-specific antigens,then establish the ELISA detection method for dengue virus antibody.Methods Using bioinformatic software DNAStar and ANTHEPROT to analyze the hydrophilicity,flexibility,surface probability and antigenicity of dengue virus type 1-4,Japanese encephalitis virus and Yellow fever virus M,E and NSI protein amino acid sequence and also consider the influence of secondary structure.Then in accordance with epitopes localion and amino acid sequence similarity,forecast the share and specific epitopes.Reference the sequence information of different dengue virus strains in GenBank to analyze the epitopes conservative.Based on the results of bioinformatic analysis,5 specific epitopes were amplified and inserted into prokaryotic expression vector pMAL-C2x or pET32a.Then the vectors was transferred into E.coli Rosetta( DE3 ).lsopropyl-β-D-thiogalactoside(IPTG) was used to induce the expression of gene segments.SDS-PAGE were used to identify the expression proteins,and the antigenicity were tested using Western blot.Using the antigen selected by Western blot,ELISA method for dengue virus antibody detection was established.Results Eighty shared epitopes and 25 specific epitopes were forecasted,and 5 antigenic fragments encloude analyzed epitopes from dengue virus type 2 and 3 were expressed in E.coli successfully.One dengue virus type 1-4 shared antigens (Den-Ag5),one dengue virus type 2 and 4 shared antigens( DenAg3),one dengue virus type 1-3 shared antigens(Den-Ag2) and two dengue virus type 1,2 and 4 shared antigens( Den-Ag1,Den-Ag4)were conformed using Western blot.Using antigens Den-Ag5,Den-Ag1 and DenAg2,the ELISA method for dengue virus antibody detection were established.Conclusion Based on the bioinformatic analysis and Western blot verification,5 dengue virus specific antigen were conformed,and the ELISA detection method for dengue virus antibody were established.

Objective To compare the effect of silica-extraction method and Silico membrane based method in DNA purification from bones and teeth.Methods DNA samples were purified respectively with the silica-extraction method and MinElute PCR Purification kit from 6 bones and 8 teeth,then tested STR types by GlobalFiler? kits. And evaluated the two methods with the success rate and the peak height. Results Both of the two purification methods can successfully obtain the STR markers of the 14 samples. And there was no statistical difference between the two methods in the average peak height from bones and teeth. Conclusion The Silico membrane based method which have more advantages in operation is an efficient method to purify DNA from bones and teeth, and there is no significant difference compared with the silica-extraction method. But the cost is higher. It can be selectively used in forensic practice.

Purpose To investigate the expression of estrogen receptor ( ER) and progesterone receptor ( PR) in endometrial tissue in patients with intrauterine adhesion ( IUA) and their clinical significance. Methods Two experimental methods, immunohistochemical MaxVision two step method and real-time fluorescencequantitative PCR ( qRT-PCR) , were used for detection of ER and PR in endome-trial tissue both in group IUA ( study group) and non IUA group ( control group) . Results MaxVision immunohistochemical method showed that ER protein expression in the study group (3. 52 ± 0. 71) was significantly higher than that (2. 75 ± 1. 00) in the control group (P=0. 01), ER mRNA expression by qRT-PCR was also significantly higher in the study group (1. 59 ± 0. 26) than that (1. 00 ± 0. 19) in the control group (P=0. 00). The immunohistochemical detection of showed that PR protein expression in the study group (3.26 ±0.70) had no significant different from that (3.58 ±0.28) in the control group (P=0.12), qRT-PCR also showed that ER mRNA in the study group (1.15 ±0.21) had no significantly different from that (1.00 ±0.31) in the control group (P=0. 21 ) . Immunohistochemical expression of ER and PR proteins showed a significant difference between the endometrial glands and en-dometrial stroma (χ2 =5. 797, P=0. 016,χ2 =4. 857, P=0. 027). Conclusion The expression of ER in endometrial tissue in the study group is higher than that in the control group, but the expression of PR has no different between the two groups, ER and PR pro-teins expression are higher in endometrial glands than that in endometrial stroma. These provide a theoretical reference for clinical use of estrogen and progesterone to IUA patients.

Objective To compare the therapeutic effect of video-assisted thoracoscopic surgery and traditional thoracotomy in fixation of traumatic multiple rib fractures.Methods Clinical data of 56 patients with traumatic multiple rib fractures treated surgically between July 2005 and September 2012 were analyzed retrospectively.Based on the treatments,the patients were assigned to video-assisted thoracoscopy group (thoracoscopy group,n =27) and traditional thoracotomy group (thoracotomy group,n =29).A comparison was done on the variables including operation time,intraoperative blood loss,ventilator support rate,duration of mechanical ventilation,length of ICU stay,incidence of lung infections,visual analogue scale (VAS) at day 3 postinjury and mortality between the two groups.Results Operation time [(128.9 ± 21.1) min vs (140.7 ± 24.2) min],ventilator support rate (70％ vs 76％) and mortality (4％ vs 7％) in thoracoscopy group revealed no statistical differences compared with thoracotomy group (P ＞ 0.05),but intraoperative blood loss [(321.1 ± 30.1)ml vs (438.1 ± 43.2)ml],duration of mechanical ventilation [(4.3 ± 2.1) d vs (7.2 ± 1.6) d],length of ICU stay [(5.9 ± 21.1) d vs (8.5 ± 1.7) d],incidence of lung infection (33％ vs 90％),and VAS [(7.0 ± 1.4) points vs (8.3 ± 0.9) points] were significantly reduced in thoracoscopy group than in thoracotomy group (all P ＜ 0.01).Conclusion Video-assisted thoracoscopic surgery is characterized by fewer intraoperative bleeding,shorter duration of mechanical ventilation and ICU stay,and lower lung infection rate during treatment of traumatic multiple rib fractures compared to traditional thoracotomy.

Purpose　To investigate the clinicopathological and immunohistochemical features of Epstein-Barr virus-associated and Epstein-Barr virus not-associated primary intestinal T-cell lymphomas(ITCL) and to study their cell origins. Methods　In situ hybridization for EBER1/2 and immunohistochemical staining for immunophenotypes, LMP-1,TIA-1,bcl-2 and CD21 were performed in 32 cases. The clinical data were analyzed and all patients were followed-up. Results　(1) In 27 of the 32 cases, EBER1/2 were detected in the tumor cells, in which 11 presented LMP-1 positive reactions. (2) All 32 cases of ITCL revealed CD45RO positivity，in which 4（12.5%） expressed CD8，8（25.0%）expressed CD4, 9（28.1%）expressed CD56,and 31（96.9%）expressed TIA-1. There were 17（53.7%）cases with CD4-，CD8-，CD56- immunophenotype. None expressed bcl-2 and CD21. 32 ITCL were classified into pleomorphic medium and large cell(n=28), monomorphic medium-sized(n=2), pleomorphic small cell(n=2). Clinically, most patients with ITCL were young males with abdominal pain, hematochezia, fever and weight loss. The prognosis of patients with ITCL showed poor (survival median was 1.7 month). (3) The differences between EBV-associated and EBV not-associated ITCL lay in hematochezia, fever and the expression of CD3, CD8 and CD56. Conclusion　Most of Chinese ITCL are EBV-associated ones with unusual clinicopathological and immunohistochemical features，which are of different lineages of T-cell subtypes, including cytotoxic T-cell or NK cell.

Aim To investigated the possible effect of COX-2 on the BMP9-induced activation of PI3K/Akt signal in progenitor cells.Methods The activity of alkaline phosphatase(ALP) was measured using histochemical staining or chemiluminescence.The mRNA level of ALP was determined using real-time PCR assay.The protein levels of osteopontin(OPN), osteocalcin(OCN), COX-2, Akt1/2 and phosphorylated Akt1/2 were detected by Western blot.The mRNA level of COX-2 was assayed with RT-PCR, and the mineralization was measured with Alizarin Red staining.Results The ALP activity was apparently increased by BMP9 in C2C12 cells, as well as the protein level of OPN and OCN.The mineralization was also markedly induced by BMP9 in C2C12 cells.BMP9 increased the level of phosphorylated Akt1/2 greatly, although no substantial effect was observed on total protein level of Akt1/2.The BMP9-induced ALP activity was dramatically decreased by the inhibitor of PI3K.The mRNA and protein level of COX-2 were both increased by BMP9 in C2C12cells, and the BMP9-induced ALP activity and mineralization were greatly attenuated by the inhibitor of COX-2.The BMP9-induced phosphorylation level of Akt1/2 was increased by the exogenous expression of COX-2, but decreased by the inhibitor of COX-2.Conclusion Activation of PI3K/Akt signaling may be a critical event in BMP9-induced osteogenic differentiation, and this process may be mediated by the BMP9-upregulated COX-2 in stem cells at least.

Objective The aim of this study was to investigate mutations of 41 STR loci. Methods 4546 bloodstain samples were typed from 1932 father–mother–child trios by using AGCU_21+1, AGCU_EX22 and GlobalFiler_ExpressTM amplification Kit. Calculate the mutation rates of STR loci. Results 154 mutations were identified at 32 of the 41 loci. The average mutation rate was 1.0×10-3per locus(95%CI: 0.8~1.1×10-3), and the mutations of SE33 was highest. 152(98.7%) mutation events were one-step mutation, 2(1.3%) events were two-steps. The mutation events occurred in 150 father–mother–child triplets. The mutations in 146(97.3%) triplets occurred at single locus, 8 mutations were observed at two loci in 4(2.7%) triplets simultaneously. 104 paternal and 22 maternal mutations could be determined under 79212 paternal and maternal allelictransfers. The ratio of paternal versus maternal mutations was 4.7:1, and 28 unassigned mutations were observed. Conclusion STR mutation are common in paternity testing, and we should pay more attention to it.

Aim To investigate the role of TGF-β3 in the anti-proliferation effect of ursolic acid(UA)in co-lon cancer cells and the possible molecular mechanism underlying this effect.Methods We introduced crys-tal violet staining,flow cytometry and Western blot as-say to determine the effect of UA on proliferation and apoptosis in HCT1 1 6 cells.The levels of TGF-β3, Smad2 /3 and β-catenin in HCT1 1 6 cell were evaluated by RT-PCR and Western blot.Finally,TGF-β3 inhibi-tor and recombinant adenovirus,and luciferase reporter assay were used to analyze the possible mechanism through which TGF-β3 mediated the anti-cancer effect of UA in HCT1 1 6 cells.Results UA inhibited the proliferation and induced apoptosis apparently in HCT1 1 6 cells.UA down-regulated TGF-β3 both in mRNA and in protein level.Meanwhile,UA decreased the phosphorylation of Smad2 /3 concentration depend-ently,although no significant effect was found on the total protein level of Smad2 /3 in HCT1 1 6 cells.Over-expression of TGF-β3 attenuated the inhibitory effect of UA on the proliferation of HCT1 1 6 cells,while the TGF-β3 inhibitor potentiated this effect. UA sup-pressed the transconduction of Wnt/β-catenin signaling in HCT1 1 6 cells through decreasing the level of β-catenin.Exogenous expression of TGF-β3 increased the level of β-catenin and partly reversed the UA-in-duced decrease of β-catenin.However,TGF-β3 inhib-itor potentiated the inhibitory effect of UA on β-catenin in HCT1 1 6 cells.Conclusion The anti-proliferation activity of UA in colon cancer may be partly mediated through down-regulating TGF-β3 to suppress Wnt/β-catenin signaling at least.

Objective:To investigate the associations between serum uric acid levels during the third trimester of pregnancy and risks of adverse pregnancy outcomes.Methods:In this retrospective study, a cohort of 7 995 pregnant women who were hospitalized for childbirth from January 2014 to January 2019 were collected to compare pregnancy outcomes between subjects with or without hyperuricemia (HUA). A smooth curve analysis was used to evaluate the relationship between uric acid levels and preterm delivery, low birth weight and smaller than gestational age. Logistic regression analyses were performed to identify risk factors for adverse pregnancy outcomes, and the interaction of the factors.Results:During the third trimester of pregnancy, the uric acid levels of about 10% pregnant women were over 420 μmol/L. In those with HUA, the median neonatal birth weight was 2 590 (1 790, 3 410) g, the probability of premature birth was 49.81%, and the incidence of small than gestational age was 20.41%. These were significantly different from the women without HUA (the median neonatal birth weight: 3300 (2850, 3640) g; the probability of premature birth 23.09%; the incidence of small than gestational age 6.55%, respectively) (All P<0.001). Maternal uric acid levels were negatively correlated with neonatal birth weight, and positively correlated with the risk of smaller than gestational age. It has a U-shaped association with the probability of premature birth, and the lowest probability of premature birth was at 200-400 μmol/L of the uric acid. Risks of low birth weight (adjusted β=-5.22, 95% CI-6.46—-3.99) and smaller than gestational age (adjusted OR=1.03, 95% CI 1.02-1.04) were increased in the function of uric acid levels. High uric acid, hypertension, oligoamnios and preeclampsia were important risk factors for the adverse pregnancy outcomes. The risk of preterm delivery and low birth weight enhanced when hyperuricemia combined with hypertension and preeclampsia. Conclusions:Serum uric acid level can be used as one of reliable markers for predicting adverse pregnancy outcomes, which might provide theoretical basis for clinical intervention in practice.