Gonadal somatic cells are the main players in gonad development and are important for sex determination and germ cell development.Here,using a time-series single-cell RNA sequencing(scRNA-seq)strategy,we analyzed fetal germ cells(FGCs)and gonadal somatic cells in human embryos and fetuses.Clustering analysis of testes and ovaries revealed several novel cell subsets,including POU5F1+SPARC+FGCs and KRT19+somatic cells.Furthermore,our data indicated that the bone morphogenetic protein(BMP)signaling pathway plays cell type-specific and develop-mental stage-specific roles in testis development and promotes the gonocyte-to-spermatogonium transition(GST)in late-stage testicular mitotic arrest FGCs.Intriguingly,testosterone synthesis function transitioned from fetal Sertoli cells to adult Leydig cells in a stepwise manner.In our study,potential interactions between gonadal somatic cells were systematically explored and we identified cell type-specific developmental defects in both FGCs and gonadal somatic cells in a Turner syndrome embryo(45,XO).Our work provides a blueprint of the complex yet highly ordered devel-opment of and the interactions among human FGCs and gonadal somatic cells.

BACKGROUND: Albuvirtide is a once-weekly injectable human immunodeficiency virus (HIV)-1 fusion inhibitor. We present interim data for a phase 3 trial assessing the safety and efficacy of albuvirtide plus lopinavir-ritonavir in HIV-1-infected adults already treated with antiretroviral drugs. METHODS: We carried out a 48-week, randomized, controlled, open-label non-inferiority trial at 12 sites in China. Adults on the World Health Organization (WHO)-recommended first-line treatment for >6 months with a plasma viral load >1000 copies/mL were enrolled and randomly assigned (1:1) to receive albuvirtide (once weekly) plus ritonavir-boosted lopinavir (ABT group) or the WHO-recommended second-line treatment (NRTI group). The primary endpoint was the proportion of patients with a plasma viral load below 50 copies/mL at 48 weeks. Non-inferiority was prespecified with a margin of 12%. RESULTS: At the time of analysis, week 24 data were available for 83 and 92 patients, and week 48 data were available for 46 and 50 patients in the albuvirtide and NRTI groups, respectively. At 48 weeks, 80.4% of patients in the ABT group and 66.0% of those in the NRTI group had HIV-1 RNA levels below 50 copies/mL, meeting the criteria for non-inferiority. For the per-protocol population, the superiority of albuvirtide over NRTI was demonstrated. The frequency of grade 3 to 4 adverse events was similar in the two groups; the most common adverse events were diarrhea, upper respiratory tract infections, and grade 3 to 4 increases in triglyceride concentration. Renal function was significantly more impaired at 12 weeks in the patients of the NRTI group who received tenofovir disoproxil fumarate than in those of the ABT group. CONCLUSIONS: The TALENT study is the first phase 3 trial of an injectable long-acting HIV drug. This interim analysis indicates that once-weekly albuvirtide in combination with ritonavir-boosted lopinavir is well tolerated and non-inferior to the WHO-recommended second-line regimen in patients with first-line treatment failure. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT02369965; https://www.clinicaltrials.gov.Chinese Clinical Trial Registry No. ChiCTR-TRC-14004276; http://www.chictr.org.cn/enindex.aspx.
Adult ; Anti-HIV Agents/adverse effects* ; Antiretroviral Therapy, Highly Active ; China ; Drug Therapy, Combination ; HIV Infections/drug therapy* ; HIV-1 ; Humans ; Maleimides ; Peptides ; Ritonavir/therapeutic use* ; Treatment Outcome ; Viral Load

Objective To compare the compressed sensing (CS)and parallel imaging (PI)techniques applied to contrast-enhanced MRI (CE-MRI)scanning of liver and to determine their clinical applicability.Methods Thirty patients with liver mass who underwent the CE-MRI scanning with both CS and PI techniques were recruited in the current study.The SNR of the liver,acquisition time and subjective image quality scores were compared between CS (CE-MRI with CS)and PI (CE-MRI with PI)groups respectively.Results The SNR values of pre-enhancement T1 WI in CS group were lower than those in PI group (1 97.82±32.5 3 vs 204.94±35.28,P＜0.05).However,there was no significant difference in the SNR values of images in equilibrium phase between the two groups (CS vs PI:392.38±72.93 vs 405.03±82.09,P＞0.05).The acquisition time in CS group was significantly shorter than that in PI group [(11.71±0.23)s vs (17.85±0.42)s, P＜0.01].Significantly higher subjective image quality scores were found in CS group than those in PI group (3.54±0.57 vs 2.91±0.80,P＜0.01). Conclusion CS technique may benefit the patients who cannot hold breath well and improve the CE-MRI image quality.

The purpose of this study is to investigate the effect of the oral endothelin antagonist Bosentan on blood pressure and renal sympathetic nerve activity (RSNA) in rats exposed to chronic intermittent hypoxia (CIH), and to explore the sympathoexcitation mechanism of endothelin-1 (ET-1) in CIH-induced hypertension. Twenty-four male SD rats were randomly divided into normoxia, CIH and Bosentan groups. Rats in the normoxia group were exposed to normoxic environment, and rats in CIH or Bosentan group were exposed to intermittent hypoxia for 3 weeks. Bosentan was given at 50 mg/kg by intragastric administration before intermittent hypoxia exposure in Bosentan group. Systolic blood pressure (SBP) was measured by BP-2000, and the change of RSNA to sodium nitroprusside (SNP) or phenylephrine (PE) was recorded by PowerLab signal acquisition system. Serums of all rats were collected and the contents of ET-1 and norepinephrine (NE) were measured by ELISA. Results showed that blood pressure was gradually increased following CIH exposure compared with the normoxia group during the 3 weeks (P < 0.01, P < 0.01, P < 0.001). The basal RSNA was increased and baroreflex sensitivity was decreased in rats exposed to CIH. Furthermore, the blood pressure was positively correlated with the level of ET-1 in serum in rats exposed to CIH (r = 0.833, P = 0.01). Bosentan administration significantly decreased SBP and basal RSNA, increased the baroreflex sensitivity, and decreased serum NE level in rats exposed to CIH. These results suggest that ET-1 is related with blood pressure elevation in rats exposed to CIH, and Bosentan reverses CIH-induced hypertension by decreasing RSNA.

BACKGROUNDThe domestic prevalence of chronic hepatitis B (CHB) in China is 7.18% in 2006, imposing great societal healthcare burdens. Nucleot(s)ide analogues (NUCs) anti-hepatitis B virus (HBV) therapies are widely applied despite the relatively low rate of seroconversion and high risk of drug-resistant mutation. More effective treatments for CHB deserve further explorations. Combined therapy of NUCs plus Chinese herbal medicine (CHM) is widely accepted in China, which is recognized as a prospective alternative approach. The study was primarily designed to confirm the hypothesis that Tiaogan-Yipi Granule (, TGYP) or Tiaogan-Jianpi-Jiedu Granule (, TGJPJD) plus entecavir tablet (ETV) was superior over ETV monotherapy in enhancing HBeAg loss rate.

METHODSThe study was a nationwide, large-scale, multi-center, double-blind, randomized, placebo-controlled trial with a designed duration of 108 weeks. A total of 16 hospitals and 596 eligible Chinese HBeAg positive CHB patients were enrolled from November 2012 to September 2013 and randomly allocated into 2 groups in 1:1 ratio via central randomization system: experimental group (EG) and control group (CG). Subjects in EG received CM formulae (TGYP or TGJPJD, 50 g per dose, twice daily) plus ETV tablet (or ETV placebo) 0.5 mg per day in the first 24 weeks (stage 1), and CHM granule plus ETV tablet (0.5 mg per day) from week 25 to 108 (stage 2). Subjects in CG received CHM Granule placebo plus ETV tablet (0.5 mg per day) for 108 weeks throughout the trial. The assessments of primary outcomes (HBV serum markers and HBV-DNA) were conducted by a third-party College of American Pathologists (CAP) qualified laboratory. Adverse effects were observed in the hospitals of recruitment.

DISCUSSIONThe study was designed to compare the curative effect of CM plus ETV and ETV monotherapy in respect of HBeAg loss, which is recognized by the European Association for the Study of the Liver as "a valuable endpoint". We believe this trial could provide a reliable status for patients' "journey" towards durable responses after treatment discontinuation. The trial was registered before recruitment on Chinese Clinical trial registry (No. ChiCTR-TRC-12002784, Version 1.0, 2015/12/23).

Objective To discuss the effects of lidocaine infusion on perioperative immune function by evaluating the levels of stress hormone and natural killer (NK) cell cytotoxicity.Methods Thirty-five patients of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,aged 35-65 yr,undergoing elective radical hysterectomy,were randomized into lidocaine group (group L)and control group (group C).Fifteen minutes before anesthesia induction,a bolus of 1.5 mg/kg of lidocaine was administered iv.to each patient in group L and followed by a continuous infusion at 1.5 mg·kg-1 ·h-1 lasting to the end of surgery.Meanwhile,the patients in group C received the same volume of saline.Venous blood samples were collected individually 24 h before the operation,the end of the operation and 48 h after the operation.Levels of prostaglandin,epinephrine and norepinephrine were assayed by ELISA kits.NK Cells were obtained by CD56 antibody magnetic isolation.The cytotoxicity of NK cell was detected by LDH releasing assay,and phosphor-protein kinase A (p-PKA)and protein kinase A (PKA) were detected by Western blotting.Results There were no significantly different in the plasm levels of PGF2,EP1 and NE.The plasm levels of prostaglandin (562.5±98.2 vs.663.2±119.0) pg/ml,epinephrine (24.9±4.8 vs.29.7±3.5) pg/ml and norepinephrine (408.3 ±47.2 vs.499.6±45.6) pg/ml in patients of group L were lower than those in group C (P＜0.05)48 h after the surgery.The cytotoxicity of NK cell was higher in group L than that in group C (44.1 ±5.0 vs.37.1±5.5)％ (P＜0.05) 48 h after the surgery.The ratio of p-PKA/PKA was lower in group L than that in guoup C (0.060±0.008 vs.0.099±0.011) (P＜0.05) at the end of the surgery.Conclusion Perioperative intravenous lidocaine infusion can reduce the level of plasma catecholamine and PGE2,and protect the cytotoxicity of NK cell,possibly via inhibiting of cAMP-PKA signaling pathway.

Objective To investigate the influencing factors for non/low-response to hepatitis B vaccine in infants of HBsAg-positive mothers.Methods A total of 286 HBsAg-positive pregnant women and their infants were recruited from the Third People's Hospital of Taiyuan during July 2011 to January 2013.The infants were immunized with hepatitis B vaccine according to the 0-1-6 month vaccination schedule and followed up for 12 months.The serum HBV DNA level of mothers,neonates and infants were detected by electro chemilum inescence immunoassay kits and fluorescene quantiative polymerase chain rection.Results Among 286 infants,the rate of non/low-response to hepatitis B vaccine was 18.53％ (53/286).Non-conditional logistic regression analysis indicated that the mother's HBV DNA level ≥ 1 × 107 copies/ml (0R=2.592,95％CI:1.121-5.996) and natural birth (OR=1.932,95％CI:1.021-3.654) were the risk factors for non/low-response to hepatitis B vaccine,the risks were 2.592 times and 1.932 times higher compared with the infants whose mothers were HBV DNA negative and the infants whose mothers had cesarean delivery.There was no multiplicative or additive interaction between high HBV DNA load and natural birth (OR=1.055,95％CI:0.209-5.321),(RERI=1.617,95％CI:-4.038-7.272;AP=0.364,95％CI:-).527-1.225;SI=1.195,95％CI:0.270-13.135).After stratified analysis of mother's HBV DNA level,delivery mode of mothers was not associated with non/low-response of their infants.Conclusion The mother's load of HBV DNA ≥ 1 × 107 copies/ml might be the factor for non/low-response to hepatitis B vaccine in infants of HBsAg positive mothers.

Objective To explore the effect of interleukin-6 (IL-6) and Interleukin-12(IL-12) on immune response to hepatitis B vaccination in infants of HBsAg-positive mothers.Methods A total of 91 neonates whose mothers were HBsAg-positive were included and followed up for 12 months.HBV DNA and HBV serological markers in the peripheral blood of the neonates and infants were detected with fluorescence quantitative polymerase chain reaction (FQ-PCR) and chemiluminescence immunoassay (CLIA),and the levels of IL-6 and IL-12 in the peripheral blood of the neonates and infants were detected with enzyme-linked immunosorbent assay (ELISA).Results The non-/hypo-response rate to hepatitis B vaccination was 35.16％ (32/91) in the 91 infants.In the neonatal period and infantile period,the level of IL-6 in non-/hypo-response group was lower than that in high-response group,while the level of IL-12 was higher than that in high-response group,and there was significant difference (P＜0.01).From the neonatal period to the infantile period,the level of IL-6 increased,while the level of IL-12 descended in both groups,and there was significant difference (P＜0.01).Furthermore,the level of anti-HBs of infants was positively correlated with the level of IL-6 (rs =0.70,0.79,P＜ 0.01),and was negatively correlated with the level of IL-12 (rs=-0.71,-0.72,P＜0.01) in the neonatal period and the infantile period.From the neonatal period to the infantile period,the increased level of IL-6 was positively associated with the level of anti-HBs (rs =-0.74,P＜0.01),while the decreased level of IL-12 was negatively associated with the level of anti-HBs (rs=-0.42,P＜0.01).The level of IL-6 was negatively correlated with the level of IL-12 in the neonatal period and the infantile period (rs=-0.68,-0.70,P＜0.01).Conclusions IL-6 might promote the immune response to hepatitis B vaccination in infants whose mothers were HBsAg-positive,while IL-12 might inhibit the immune response.IL-6 and IL-12 would affect the immune response to hepatitis B vaccination in infants of HBsAg-positive mothers at the same time.

Objective To study the expression and significance of Ezrin in triple negative breast cancer tissues .Methods We selected 102 cases ,including 24 ones of triple negative breast cancer ,58 ones of non‐triple negative breast cancer ,and 20 of benign breast disease .The expression of Ezrin in all the specimens was detected by SP immunohistochemistry .We observed whether there was any difference between the positive expression rates of Ezrin in the three groups . We also analyzed the correlation between Ezrin expression and clinicopathologic parameters of triple negative breast cancer .Results The positive expression rate of Ezrin in groups of triple negative breast cancer , non‐triple negative breast cancer , and benign breast disease was 15 .00% ,48 .28% and 75 .00% ,respectively . The difference between the three groups differed significantly ( P < 0 .01 ) . The high expression of Ezrin in triple negative breast cancer had relationship with histological grading and clinical TNM staging (P< 0 .05 ) , but not with patients' age , tumor size , or axillary lymph node metastasis ( P > 0 .05 ) . Conclusion Ezrin is highly expressed in triple negative breast cancer tissues ;therefore , it can be used as an important indicator of poor prognosis of triple negative breast cancer .

BACKGROUNDIn qualitative diagnosis of bile duct stenosis, single diagnostic measure is difficult to make a correct diagnosis, to combine several diagnostic techniques may be helpful to make an accurate diagnosis. The aim of this study was to evaluate the value of intraductal ultrasonography (IDUS), endoscopic brush cytology and K-ras, P53 gene mutation in the early diagnosis of malignant biliary stricture.

METHODSFrom February 2012 to February 2013, 84 patients with suspected malignant biliary stricture were performed IDUS firstly, then endoscopic brush cytology and finally K-ras, P53 gene mutation detection, the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of all above ways were evaluated and compared.

RESULTSOf 84 patients, 52 cases were ultimately diagnosed malignant biliary stenosis; of which, 9 cases had no recurrence or metastasis to other organs after radical operation during the follow-up period. IDUS combined with brush cytology and K-ras + P53 gene mutation detection had obvious advantage in the sensitivity, accuracy and negative predictive value than any other joint detection and single detection (the advantage was more significant compared with IDUS + brush cytology or any single detection P < 0.01). There were obvious statistical significance in the sensitivity and accuracy between IDUS + brush cytology + P53 or IDUS + brush cytology + K-ras and IDUS + brush cytology or IDUS (P < 0.05). There was no statistical significance in the sensitivity, specificity, positive predictive value, negative predictive value and accuracy between IDUS + brush cytology + P53 and IDUS + brush cytology + K-ras (P > 0.05).

CONCLUSIONSIDUS combined with brush cytology and K-ras, P53 gene mutation detection is better than the separate detection and contribute to the early diagnosis of malignant biliary stricture. Its more widespread use is recommended.

Aged ; Aged, 80 and over ; Bile Duct Diseases ; diagnosis ; genetics ; Bile Duct Neoplasms ; diagnosis ; genetics ; Bile Ducts ; pathology ; Constriction, Pathologic ; diagnosis ; genetics ; Female ; Genes, p53 ; genetics ; Genes, ras ; genetics ; Humans ; Male ; Middle Aged ; Mutation