Objective To investigate the effect of dexamethasone combined with clomiphene on serum sex hormone levels in patients with polycystic ovary syndrome. Methods 80 pregnancies with polycystic ovary syndrome treated in our hospital from January 2013 to December 2015 were selected and they were randomly divided into observation group and control group with 40 cases in each group. The observation group was given dexamethasone combined with clomiphene, while the control group was given simple clomiphene. Compared the curative effect and the change of the levels of progesterone and estradiol before and after treatment for three months. Results After treatment, the total effective rate of the observation group was 95.00%, higher than the total effective rate of the control group was 72.50%, the difference was statistically significant (P<0.05), compared the levels of progesterone(P), estradiol(E2), luteinizing hormone(LH) and LH/FSH in the observation group and the control group, the difference was statistically significant (P<0.05), after treatment, both levels were higher than before treatment. The total ovulation rate of the observation group was 90.00%, higher than the total ovulation rate of the control group was 72.50%, the difference was statistically significant (P<0.05). The level of progesterone and estradiol in serum of polycystic ovary syndrome treated by dexamethasone combined with clomiphene had a obvious positive correlation (r=0.746, 0.812, 0.731 and 0.796, P<0.05). Conclusion Dexamethasone combined with clomiphene treatment of polycystic ovary syndrome than using only clomiphene treatment is good, which can also improve serum hormone levels in patients, effectively improve the patient's endocrine abnormalities.

Humans ; Adolescent ; DNA/genetics* ; Genotype

Background and purpose: With the development of therapy equipments and technology, the treatment for nasopharyngeal carcinoma(NPC) has entered into the era of precision radiotherapy, and setup errors have become a very important factor affecting treatment effects. The purpose of this study was to analyze the set-up errors detected by the kilovoltage cone beam CT(EPID) and the megavoltage electronic portal imaging device(CBCT) using 2 kinds of different immobilization techniques (pillow+head neck shoulder mask and vacuum bag+head neck shoulder mask) for NPC patients. Methods:A total number of 40 NPC patients were randomly assigned into 2 groups (pillow+head neck shoulder mask group and vacuum bag+neck shoulder mask group). Then each group was further divided into CBCT scan group and EPID group for veriifcation before treatment delivery. We matched the EPID images with the DRRs and acquired the set-up errors in x, y, z axis. Setup errors of CBCT were calculated according to its matched and planned CT images in left-right (x), superior-inferior (y) and anterior-posterior (z) directions. Paired t-test was used to evaluate the differences. Results:In the pillow+head neck shoulder mask group, the set-up errors of CBCT in the x, y, z axis were x (0.67±2.01)mm, y (0.51±1.71)mm and z (0.57±2.04)mm, respectively. The errors of EPID were x (0.69±2.19)mm, y (0.54±2.03)mm and z (0.61±2.11)mm. In the vacuum bag+head neck shoulder mask group, the set-up errors of CBCT in the x, y, z axis were x (0.42±1.81)mm, y (0.33±1.55)mm and z (0.50±1.75)mm, respectively. The errors of EPID were x (0.44±1.87)mm, y (0.43±1.70)mm and z (0.54±1.77)mm. The vacuum bag+head neck shoulder mask ifxed technique was more accurate when compared to the pillow + head neck shoulder mask ifxation method (P<0.05). Conclusion:CBCT and EPID were similar in detecting set-up errors for the NPC patients. However, the vacuum bag+neck shoulder mask ifxed technique was more accurate when compared to the pillow+head neck shoulder mask ifxation method.

Objective To evaluate the value of immature granulocytes count in diagnosing and monitoring the systemic inflam-matory response for patients with systemic inflammatory response syndrome,and to provide a new indicator of systemic inflammato-ry response.Methods 207 patients suspected of systemic inflammatory response syndrome were enrolled.The dynamic changes of immature granulocytes counts and the disease situation were recorded for all subjects.The blood samples were collected in vacuum tubes with EDTA-K2 anticoagulant.Blood cell count and immature granulocytes count were performed in Sysmex XE-2100 hema-tology analyzer.The determination of C-reactive protein and procalcitonin were also completed.The performance of immature granu-locytes in diagnosing systemic inflammatory response syndrome was evaluated by receiver operating characteristic curve analysis. Results The area under the curve of immature granulocyte count (IG #)was 0.78 in diagnosing systemic inflammatory response syndrome,with a sensitivity of 62.2 % and a specificity of 73 % at IG #> 0.1 65.The area under the curve of immature granulo-cyte percent (IG%)was 0.771 in diagnosing systemic inflammatory response syndrome,with a sensitivity of 54.1 % and a specifici-ty of 94.6 % at IG%>2.55 %.The area under the curve of C-reactive protein was 0.71 6 in diagnosing systemic inflammatory re-sponse syndrome,with a sensitivity of 67.6% and a specificity of 75.7 % at C-reactive protein> 64.15 mg/L.The area under the curve of procalcitonin was 0.772 in diagnosing systemic inflammatory response syndrome,with a sensitivity of 75.7 % and a speci-ficity of 70.3 % at procalcitonin> 0.33 mg/L.Conclusion Immature granulocyte count is beneficial for the diagnosis and the as-sessment of prognosis of systemic inflammatory response syndrome.

Objective: To analyze the epidemic trend of viral hepatitis in Nanjing from 1989 to 2019 and predict the incidence in 2020, so as to provide reference for the prevention and control of viral hepatitis. Methods: The incidence data of viral hepatitis in Nanjing from 1989 to 2019 was retrieved from Nanjng Center for Disease Control and Prevention and National Infectious Disease Reporting System. The epidemic trend was analyzed by estimating the annual percent change ( APC ) and the average annual percent change ( AAPC ). The seasonal incidence of different types of viral hepatitis was analyzed by seasonal index. The autoregressive integrated moving average model ( ARIMA ) was built to predict monthly incidence rate of viral hepatitis in 2020. Results: The annual incidence rate of viral hepatitis was 62.00/100 000 in Nanjing from 1989 to 2019, showing a downward trend ( AAPC=8.4%, P<0.05 ). From 1998 to 2019, the annual incidence rates of hepatitis A, B, C and E were 1.98/100 000, 14.31/100 000, 2.30/100 000 and 2.60/100 000. The incidence of hepatitis A and B showed downward trends ( AAPC=-11.81%, -6.02%, both P<0.05 ); the incidence trend of hepatitis C was not obvious ( P>0.05 ); the incidence of hepatitis E showed an increasing trend ( AAPC=4.82%, P<0.05 ). From 2015 to 2019, the third and fourth quarters were the epidemic seasons of hepatitis A, B and C, while the first and second quarters were the epidemic seasons of hepatitis E. The ARIMA model predicted that the monthly incidence rates of viral hepatitis in 2020 would range from 1.26/100 000 to 3.69/100 000, among which hepatitis B ranged from 1.21/100 000 to 2.58/100 000, hepatitis C from 0.20/100 000 to 0.48/100 000, hepatitis E from 0.09/100 000 to 0.25/100 000. Conclusions The incidence of viral hepatitis in Nanjing shows a downward trend. Among different types of hepatitis, hepatitis B has a higher incidence. All types of hepatitis have epidemic seasons. It is predicted that the monthly incidence rates of viral hepatitis will be 1.26/100 000 to 3.69/100 000 in 2020.

BackgroundChondroitin sulfate(CS) is a highly viscous and elastic acid mucopolysaccharide extracted from animal soft tissues,with a wide range of biological activity for use in clinical ophthalmology.Interim preservation solution containing CS has a significant protective effect on corneal endothelial cells (CECs).However,the protective effect played by CS in long-term glycerol cryopreservation of CECs remains to be studied.Objective This study is mainly attempted to investigate the protective effect of CS on graft CECs after cryopreservation by glycerin,and to compare the preserving outcome with that of sodium hyaluronate (SH).MethodsOne hundred and four eyes of fifty-two female Wistar rats were divided into four groups randomly.The cornea grafts were evenly anointed on the surface of the endothelium by 3％ CS and 1％ SH,respectively,and then cryopreserved in glycerol in the 3％ CS group and 1％ SH group,and the corneas cryopreserved only in glycerin were assigned to the glycerin only group.The fresh corneas of matched rats were used as the normal control group.Ninety-six female SD rats were appointed as recipients to receive allogeneic penetrating keratoplasty (PKP) 2 months after the graft cryopreservation.The viability of CECs on the corneas cryopreserved for 20 days and grafts of 14 days following surgery was assessed using trypan blue & alizarin red stain.The ultrastructure of cryopreserved corneas was examined under a transmission electron microscope.The rejection index (RI) and graft survival time were evaluated by inflammatory scoring under the slit lamp 1 day after operation,and regular pathological examination was performed after 7,14 or 30 days.The expression of transforming growth factor-β1 ( TGF-β1 ) and intercellular adhesion molecule-1 ( ICAM-1 ) in corneas and grafts were detected by immunochemistry.ResultsCECs morphology was normal in fresh cornea in normal control group,presenting no blue stained cells after trypan blue staining.In the 3％ CS group,few blue-stained cells were seen:however,the number of blue-stained cells noticeably increased in the 1％ SH group and glycerin only group.The viability and density of CECs were significantly lower in the 1％ SH group and glycerin only group,compared with the 3％ CS group (P＜0.05).Edema of the endoplasmic reticulum,inflammatory infiltration and neovascularization were more severe in the 1％ SH group and glycerin only group than the 3％ CS group 14 days after PKP.Compared with the 1％ SH group and glycerin only group,the R1 of the 3％ CS group was significantly reduced and the CECs survival time was delayed after PKP ( P＜0.05 ).Immunochemistry revealed that the expression of TGF-β1 in grafts (A value)was up-regulated and that of ICAM-1 was down-regulated in the 3％ CS group,compared with the 1％ SH group and glycerin only group ( P＜0.05).ConclusionsThe combination of 3％ CS with glycerol to cryopreserve corneal donor can maintain the viability of CECs for a longer duration and improve the effectiveness of PKP.

AIM:To construct eukaryotic expression vector of Sirt2 and detect its expression in HEK293 cells. METHODS: Total RNA was isolated from brain tissue of a-dult SD rat. A 1 130 bp fragment containing the coding region of Sirt2 was amplified by RT-PCR and the resulting PCR product was subcloned into PMD20-T vector and se-quenced. Coding region of Sirt2 was generated with PCR by using the PMD20-T-Sirt2 as template, the amplified PCR fragment was inserted into the EcoR I and Hind Ⅲ sites of the pcDNA3. 1myc-his(-)A expression vector, and the sequence was confirmed by DNA sequencing. The expression of new construct pcDNA3.1 myc-his(-) A-Sirt2 in HEK293 cells was detected by immunofluorescence. RESULTS: The full length coding region of Sirt2 was obtained and confirmed by sequencing, the expression of Sirt2 was detected successfully in HEK293 cells. CONCLUSION: The eukaryotic expression vector of Sirt2 has been successfully constructed, which will provide a useful tool for designing an in-depth investigation of the role of Sirt2.

Objective To explore the preventive and therapeutic role of silencing type Ⅰ rat platelet-binding protein motifs depolymerization protein-like metalloproteinase 2(ADAMTS2)by siRNA on experimental liver fibrosis in vitro.By studying the mechanism of siRNA silencing of ADAMTS2,we also aim to evaluate the feasibility of ADAMTS2 as a target for anti-liver fibrosis therapy.Methods Three pairs of siRNAs targeting ADAMTS2 mRNA 2237,2597 and 690 targets were designed and synthesized by utilizing RNA design software.The most effective siRNA was chosen to transfect HSC-T6 cell line to test the tendency of hepatic stellate cell(HSC)activation and ex pression of ADAMTS2,COL1α1,COL(I),α-SMA,TGF-β1,MMP-2 and TIMP-3.These were quantified using real time-PCR,Western blotting,and MTT assays.Results Of the same dosage and time of injection,siRNA 2237 inhibited ADAMTS2 gene expression significantly more than other siRNAs.siRNA-ADAMTS2 2237 markedly inhibited ADAMTS2 gene and protein expression of HSCT6 with more than 80％ efficiency.Conversely,siRNA-ADAMTS2 2237 markedly reduced the gene and protein expressions of COL(I),α-SMA and TGF-β1 on HSC-T6 and inhibited the proliferation of HSC.Conclusions siRNA-ADAMTS2 2237 could effectively knockdown the gene and protein expression of ADAMTS2 in HSC-T6 cell lines.Silencing ADAMTS2 by siRNA significantly inhibited the activation,proliferation of HSC and the gene and protein expressions of COL(I),α SMA,and TGF-β1,and it may have a potential anti-fibrotic effect.ADAMTS2 might be an efficient target for anti-fibrotic therapy.

Objective To explore the effects of inhibiting DDR2 expression by siRNA on hepatic stellate cells and evaluate the role of DDR2 gene in hepatic fibrogenesis. Methods (1) Three pairs of chemically synthesized siRNAs targeting DDR2 were respectively transfected into HSC-T6 cells for evaluation of silence efficacy, and the most effective siRNA was used. (2) HSC-T6 cells were divided into three groups, group A served as normal controls, group B served as negative control and group C was RNA interference DDR2 (siRNA-DDR2) expression of HSC. The most effective RNA interference sequences targeting DDR2 gene was chosen to transfect HSC-T6 cells by plasmid transfection. The tendency of DDR2, α-smooth muscle actin(α-SMA) and collagen-Ⅰ mRNA expression were estimated using RT-PCR, and the protein expression of DDR2 was evaluated by Western blot. Meanwhile, MTT assay was employed to analyze the proliferation of HSC. Results (1) DDR2 siRNA, which began at nt 868, inhibited DDR2 gone expression stronger than the other two siRNAs. (2) After transfection of siRNA-DDR2, the mRNA expression of DDR2 (P<0.01) and α-SMA (P<0.01) significantly decreased compared with the normal group, and the protein expression of DDR2 also significantly decreased (P<0.01). In addition, the proliferation of HSC was also markedly suppressed as compared with the normal group (P<0.01). However, compared with the negative control group, none of them was markedly suppressed. Conclusion SiRNA targeting DDR2 significantly suppresses the activation, proliferation of HSC, and thus attenuates hepatic fibrogonesis in vitro.

BACKGROUND:Immunity of fetal liver stem cel transplantation is rarely reported, syngeneic and al ogeneic fetal liver stem cel transplantation in the treatment of hepatic cirrhosis is stil unclear.
OBJECTIVE:To observe the therapeutic effects of syngeneic and al ogeneic fetal liver stem cel transplantation on hepatic cirrhosis as wel as immune rejections during the therapeutic process.
METHODS:The fetal liver stem/progenitor cel s from BALB/c and C57BL/6 mice were isolated and purified by the type IV col agen enzyme digestion method. A total of 104 healthy BALB/c mice were randomly assigned to four groups. Normal control group:no treatment;Hepatic cirrhosis group, syngeneic transplantation group and al ogeneic transplantation group:16 weeks after hepatic cirrhosis models of mice were developed by intraperitoneal injection with carbon tetrachloride, physiological saline, syngeneic fetal liver stem cel s and al ogeneic fetal liver stem cel s were injected via the caudal vein. Final y, the survival statuses, liver function, hepatic fibrosis index, the number and ratio of immune cel s (CD4+T, CD8+T, NK, NKT) and histopathologic examinations were compared in each group after transplantation 4 weeks.
RESULTS AND CONCLUSION:The survival rates in the two transplantation groups were both 100%, which was significantly higher than that in the hepatic cirrhosis group (67%, P<0.05). The liver function and liver fibrosis index in each group did not show statistical differences (P>0.05). Immunological tests showed no difference between groups (P>0.05). Pathohistology examination of hepatic tissue repair:Al ogeneic transplantation group>syngeneic transplantation group>hepatic cirrhosis group. Hence, fetal liver stem cel transplantation via the caudal vein could elevate the survival rate of hepatic cirrhosis mice, al eviate the degree of hepatocyte necrosis. There is no immunologic rejection during syngeneic and al ogeneic fetal liver stem cel transplantation that could help to treat hepatic cirrhosis in mice.