Objective To investigate the effects of glutamine (Gln) supplementation on neurologica severity score,brain edema,neuron apoptosis,and endoplasmic reticulum stress (ERS) response after traumatic brain injury (TBI) in rats.Methods TBI rat models were established using modified Feeney's method.Eighty Sprague-Dawley rats were divided into 4 groups with a random number table:sham operation group (Sham group),TBI group,Gln supplementation group (TBI + Gln group) and ERS inducer 2-deoxy-D-glucose group (TBI +Gln + 2-DG group).We measured the rats' neurobehavioral outcomes by modified neurologic severity score (mNSS) on day 1,3,7 and 14 after TBI.Neuron apoptosis was detected using TUNEL staining.Brain water content was measured with wet-dry weight method.The apoptosis-related protein (caspase-12,caspase3,and Bcl-2) and ERS-related cytokines [inositol-requiring enzyme 1 (IRE-1),C/EBP homologous protein (CHOP)] expressions in TBI cerebral cortex were determined by immunohistochemistry staining and Western blot.Results Compared with the Sham group,the levels of brain edema,mNSS,apoptosis-related protein (caspase-12,caspase-3,Bcl-2) and ERS-related proteins (IRE-1,CHOP) were significantly increased in the other three groups (all P =0.00).Compared with the TB1 group,the TBI +Gln group showed significant lower brain water content [3 d:(81.39±0.59)％ vs.(83.54±0.52)％,P=0.04;7 d:(74.86±0.38)％ vs.(77.32±0.66)％,P=0.03],improved mNSS (8.63 ±0.22 vs.10.37±0.29,P=0.03),suppressed expressions of apoptosis-and ERS-related proteins (caspase-12,caspase-3,IRE-1,and CHOP)(P =0.01,P ＜ 0.01),and increased expression of anti-apoptotic protein Bcl-2 (P =0.02).Compared withthe TBI + Gln group,the expression of ERS-related factors (IRE-1 and CHOP),brain edema level,and neurological severity were increased in the TBI + Glu + 2-DG group.Conclusion Glutamine supplementation may have neuroprotection function,demonstrated as reducing brain edema and neuron apoptosis,and improving neurobehaviroal outcomes after TBI,possibly mediated by inhibiting TBI-induced ERS response.

Objective To evaluate the efficacy of ultrasound-guided combined C5 and superficial cervical plexus block in patients undergoing clavicle surgery.Methods Sixty ASA physical status Ⅰ or Ⅱ patients,scheduled for elective clavicle surgery,were randomly divided into 2 groups (n =30 each) using a random number table:injection with 10 ml local anesthetic guided by ultrasound group (group U),and injection with 10 ml local anesthetic guided by anatomical landmark group (group A).A mixture of 0.375 ％ levobupivacaine and 1.0％ lidocaine was used.The time spent performing the block onset time of analgesia,onset time of analgesia in the medial border,midpoint and lateral border of the clavicle and duration of analgesia were measured.The effectiveness of block (excellent,good,failure) was assessed.The complications were observed.Results Compared with group A,the time spent performing the block was significantly prolonged,the onset time of analgesia in the lateral border and midpoint of the clavicle was shortened,the rate of excellent anesthesia was increased,and the duration of analgesia was prolonged in group U (P ＜ 0.05).No complications occurred in group U,while vascular puncture occurred in 3 cases,and one patient developed mild toxic reaction in group A.Conclusion Ultrasound-guided combined C5 and superficial cervical plexus block provides better block,with faster onset time of analgesia in the lateral border and midpoint of the clavicle,longer duration of analgesia and fewer complications as compared with that guided by anatomical landmarks in patients undergoing clavicle surgery.

Objective To compare the rates of regimen modification between patients with different initial antiretroviral therapy, and to investigate risk factors associated with drug toxicity-related regimen modification.Methods A two-years retrospective cohort study was conducted in 14 060 patients who initiated antiretroviral treatment with Zidovudine (AZT)/Tenofovir disoproxil (TDF)+Lamivudine (3TC)+Efavirenz (EFV) since 2012.There were 5 126 patients initiated TDF+3TC+EFV therapy (TDF group) and 8 934 patients initiated AZT+3TC+EFV therapy (AZT group).Chi-square test was used to compare the rate of first-line regimen modification and the rate of toxicity-related regimen modification between two groups.Cox proportional hazard model was used to investigate the risk factors associated with regimen modification.Results A total of 14 060 acquired immunodeficiency syndrome patients were observed for a median period of 1.85 person-years.There were 2 795 patients who changed their initial antiretroviral regimen and the rate of initial regimen modification was 19.9%.Two hundred patients who changed their initial regimen due to pregnancy were excluded.There were 2 070 patients in AZT group who changed their initial regimen with a rate of 23.5%.Among them, 1 652 patients changed their regimen due to drug toxicity and the rate was 18.8%.There were 525 patients in TDF group who changed their initial regimen with a rate of 10.4% and the rate of toxicity-related regimen modification was 6.2%.The differences between two groups were statistical significance (χ2=366.68 and 416.89, respectively, both P<0.01).The risk of regimen modification in AZT group were significantly higher than that in TDF group (aHR=2.89, 95%CI: 2.57-3.24).The risk of toxicity-related regimen modification in AZT group was also significantly higher than that in TDF group (aHR=3.85, 95%CI: 3.34-4.45).Conclusions Patients initiated antiretroviral treatment with AZT+3TC+EFV are more likely to change their initial regimen than those who initiated treatment with TDF+3TC+EFV.Female, age >45 years old, BMI<18.5 kg/cm2 and baseline CD4+ T cell count<200/mL were risk factors associated with regimen modification.

Objective To reconstruct the initiative procedure of HIV-1 reverse transcription in vitro and establish a methodology of assessing activity of HIV-1 reverse transcriptase (RT) with real-time PCR Methods The tRNALys-3 gene was amplified from genome in healthy individuals through polymerase chain reaction (PCR), and then T7 transcription promoter was added in 5'-terminal of the tRNALys-3. The tRNA[Lys-3 cRNA product was obtained by applying T7 RNA polymerase through a transcription reaction. The 5'-LTR-PBS DNA was also obtained by transcription reaction from the HIV-1 infectious clone and inserted into pGEM-T easy vectors. 5'-LTR-PBS cRNA was obtained by applying SP6 RNA polymerase whose combining site was located in pGEM-T easy vectors. Then the two RNA samples was catalyzed by two kinds of standard reverse transcriptases (SuperScript Ⅲ and HIV-1 standard reverse transcriptase, respectively) and the cDNA was synthesised. The relative activity of RT was determined with the real-time PCR. Results The tRNALys-3 primer and the SP6-5'-LTR-PBS RNA were procured accurately, whose length were 93bp and 872 bp, respectively. After the following serial dilution of Super Script Ⅲ and HIV-1 standard reverse transeriptase:1 : 10, 1: 100, 1:1 000, 1:10 000, each step of reverse transcription process worked successfully. Real-time PCR results showed that Ct values of the two groups were 13.9, 18. 3, 20. 9, 24. 9 and 20. 4, 25. 5, 28. 7, 32. 5 respectively. Conclusion A novel real-time PCR method is developed to assay the RT activity directly through reconstructing the initiation of HIV reproduction, which may be helpful for clinical management, screening of new antiretroviral drugs, and drug resistance test.

Objective To investigate the prevalence of malnutrition in human immunodeficiency virus ( HIV )‐infected children in China , and to explore and analyze the factors associated with malnutrition .Methods A cross‐sectional study was conducted by the antiretroviral treatment database of children .HIV‐infected children aged between 0 - 15 years old who initiated antiretroviral treatment were collected between January 1st , 2010 and December 31st , 2014 . Z‐score of height and weight were calculated by WHO Anthro (plus) software .Univariate and multivariate Logistic model analyses were performed to determine the factors associated with acute /chronic/mixed malnutrition .Results Baseline data of the 3 138 HIV‐infected children showed that 1 645 patients (52 .42% ) had malnutrition before antiretroviral treatment ,with acute ,chronic and mixed malnutrition of 8 .76% (275) ,39 .77% (1 248) and 3 .89% (122) ,respectively according to the type of malnutrition .Multivariate analysis showed that baseline CD4 + cell count < 200 cells/μL was the risk factor associated with acute malnutrition (aOR =2 .27 ,95% CI :1 .68 - 3 .06) ;rural settings (aOR = 1 .30 ,95% CI :1 .11 - 1 .53) ,baseline CD4 + cell count < 200 cells/μL (aOR = 1 .98 ,95% CI :1 .65 - 2 .38) ,baseline CD4 + cell count between 200 to 350 cells/μL (aOR = 1 .38 ,95% CI :1 .13 - 1 .69) and having AIDS‐related diseases (aOR = 1 .34 ,95%CI :1 .13 - 1 .59) were risk factors associated with chronic malnutrition ;and age of 11 - 15 years (aOR =2 .38 ,95% CI :1 .46 - 3 .88) ,baseline CD4 + cell count < 200 cells/μL (aOR = 4 .99 ,95% CI :3 .04 -8 .21) and having AIDS‐related diseases (aOR = 2 .45 ,95% CI :1 .65 - 3 .66) were risk factors associated with mixed malnutrition .Conclusions The prevalence of malnutrition in untreated HIV‐infected children remains high .All three types of malnutrition are associated with immunodeficiency .Early diagnosis and early treatment should be improved in HIV‐infected children through antiviral therapy to reduce the destruction of HIV to immune system .At the same time ,intensified monitoring of the nutritional status and nourishing undernourished children should be strengthened to reduce the prevalence of malnutrition .

reproducible, and may cover the major circulating strains in China.

Objective To establish a mini-pool nucleic acid testing (NAT) assay using multiplex RT-nested PCR for the detection of HIV RNA, and apply it in screening for acute HIV infection among MSM. Methods Frozen EDTA plasma samples collected between Oct. 2008 and Mar. 2009 from 3 HIV infectors during window-period, a total of 30 HIV chronically infected individuals and 97 healthy subjects were used to develop the NAT assay. Plasma samples from 10 cases were pooled into one tube and centrifuged at high speed for the collection of viruses. HIV RNA was extracted. Two pairs of primers were designed according to two conserved regions of HIV RNA ( HXB2 nt 5783-nt 6228 and nt 1235-nt 2012).Multiplex RT-PCR and nested PCR were performed. Individual NAT-reactive samples were confirmed by commercially available NAT assays. The sensitivity and performance efficacy were also evaluated. The assay was then applied to 1 005 plasma specimens from MSM with negative or uncertain HIV antibody test results.These were collected in the same period as the other samples. Results ( 1 ) Two fragments of HIV were amplified successfully with the low detection limit of 162 copies/ml plasma; (2) Results of the mini-pool HIV NAT validation with samples from 3 HIV infectors during window-period were consistent with the expected values; (3) All 30 plasma samples from MSM with positive HIV antibody, which were tested by multiplex RT nested PCR, were found to be HIV RNA positive; (4) One out of 1 005 plasma samples was found to be HIV RNA positive, for this case acute infection was followed-up and sero-conversion was found. Conclusion Mini-pool NAT has good sensitivity, and may be applied to screening HIV RNA among MSM during window-period.

Objective To study clinical features and mechanism in patients suffered from chronic hepatitis B achieving seroconversion of HBsAg by combination treatment with interferon (IFN) and nucleoside analogue (NA). Methods Thirty-two cases with chronic HBV hepatitis were enrolled into this retrospective study. All of them received combination treatment with IFN and Lamivudine/Adefovir, as well as achieved seroconversion of HBsAg from June, 2001 to May, 2007. All the cases in this study were followed up. Results Generally, serum HBV DNA fell below the detection limit 3 to 6 months after starting combination treatment. Virological breakthrough/relapse or new clinical resistant had not been found in all enrolments after combination treatment, including patients with previous resistant to Lamivudine, although the average length of treatment was over 2 years. The average period of following up after seroconversion of HBsAg was 13.2 months. Two cases transfered back to HBsAg positive, one of them achieved seroconversion of HBsAg again by the anti-virus treatment, and the other one gave up treatment and remained anti-HBe positive and HBeAg negative.The other 30 eases kept at the stage of seroconversion of HBsAg. Seven patients underwent liver biopsy after seroconversion of HBsAg, and 3 of them had taken liver biopsy before combination therapy too. Biopsy specimens were scored for fibrosis and neeroinflammation according to the Knodell histological activity index. Six cases showed HBsAg and HBcAg negative by immunohistochemistry,and only 1 case with HBsAg positive in liver tissue experienced relapse. Inflammation and fibrosis grade of the 3 cases who had taken liver biopsy twice were lowered after HBsAg seroconversion,although the ALT level of 1 case who had turned from G2S4 to GIS2-3 remained abnormal after HBsAg seroconversion. According to the sequence and character of HBsAg seroconversion, there were three models of HBsAg conversion. The sequence of transition was HBV DNA→HBeAg→HBsAg,which was dominant one, accounting for 59%(19/32 cases). HBV DNA negative, and the titer of HBeAg wandering at a low level, after then HBeAg and HBsAg change to negative in the same time,31% (10/32 cases). The titer of HBsAg decreased rapidly after the HBV DNA clearance, and the HBsAg clearance was earlier than HBeAg, 9% (3/32 cases). After 1 year of combination therapy,there were 15 of 21 cases (71.4%) whose titer HBsAg showed less than 100 COI by agent from Roche, and 7 of 11 cases (63.6%) whose titer HBsAg showed less than 250 IU/L by agent from Abbott. The frequency of adverse reaction was similar with that induced by IFN monotherapy, and no new adverse reaction was found. Conclusions Combination therapy and long course treatment might be the key to achieve the HBsAg seroconversion. Those with HBsAg in liver tissue and (or) low serum anti-HBs are more likely to relapse. The titer of HbsAg<100 COI (Roche, Germany) or<250 IU/L (Abbott, USA) after one year treatment may be regarded as a predict index of HBsAg seroconversion.

Objective To investigate the effects and mechanisms of glutamine (Gln) supplementation on oxidative stress,autophagy response and neurobehavioral outcome after traumatic brain injury (TBI) in rats.Methods TBI animal models were established using Feeney's method.Eighty SD rats were randomly divided into 4 groups:sham operation group (group Sham),Sham + glutamine supplementation group (group Sham+ GLN),traumatic brain injury group (group TBI),and TBI + glutamine supplementation group (group TBI+ GLN).We measured rat behavioral outcomes by modified neurologic severity score (mNSS) tests at day 1,3,7 and 14 after TBI.The apoptosis neurons in TBI cerebral cortex were determined by TUNEL staining.The expression of reactive oxygen species (ROS) was tested by ROS kits.Oxidative stress and autophagy related cytokines (HO-1,NQO1,Nrf2,LC3-Ⅱ and Beclin-1) were tested with Western blotting.Results Compared with the TBI group,the neurological function was improved [(9.79±0.43) vs.(8.43±0.30),F =6.775,P =0.010] and the apoptosis rate decreased (19.88％ ± 1.60％ vs.15.35％ ± 1.28％,P =0.013) in the TBI+ GLN group after 7-day treatment.Compared with the Sham group,the protein expression of ROS increased (P=0.000),and the expression of anti-oxidative stress factors (HO-1,NQO1) and Nrf2 pathway significantly decreased in the TBI group.After glutamine supplementation was given,the expression of ROS decreased and the expressions of HO-1 and NQO1 increased.The Nrf2 pathway and autophagy response also were activated with the expressions of Nrf2,LC3-Ⅱ and Beclin-1 increasing.Conclusion Glutamine supplementation can markedly reduce neuron apoptosis and improve neurological outcomes after TBI,thus has the protective effect on nerves by inhibiting TBI-induced oxidative stress response,activating Nrf2 pathway and autophagy response.

Objective: To explore the genetic basis for a case of recurrent fetal congenital hydrocephalus. Methods: Next-generation sequencing was carried out for the fetus, the gravida and two of her sisters. Results: The fetus was found to harbor a c. 1765T>C (p.Tyr589His) mutation in exon 14 of the L1CAM gene, which was derived from the gravida. Conclusion Male fetuses with recurrent hydrocephalus should be subjected to testing of the L1CAM gene to facilitate genetic counseling and prenatal diagnosis.