Objective To analyze and evaluate the current progress of blood transfusion department establishment and clinical blood transfusion management in Hangzhou,China.Methods Questionnaires were sent to 35 medical institutions to gather information regarding blood transfusion department establishment and clinical blood transfusion management in the provincial,municipal and county tier hospitals.The investigated criteria covered the establishment of the hospital blood transfusion management committee,the utilization of software and hardware of the transfusion department,the compatibility status of various software systems in use and performance evaluation of these systems in clinical applications,etc.In addition,tests were also conducted on members of the blood transfusion departments to confirm whether they are properly trained and present adequated knowledge of clinical blood applications Results were then collected for statistical and descriptive analyses.Results The results out of the 35 hospitals surveyed presented no obvious statistical significance.Nevertheless,the average score of these hospitals in different segments helped us to reach the following conclusions:1,Triple-A hospitals scored the highest average score for patient clinical status evaluation pre-transfusion;2.Double-B hospitals came first when it comes to patient clinical status evaluation post transfusion;3.Triple-B hospitals possessed the most thorough clinical records for transfusion treatment.A total of 350 papers were issued and 350 papers were collected,with 327 out of the 350 considered valid.As for the test results regarding the departments members,triple-A,triple-B and double-A hospitals scored significantly higher than double-B and unranked hospitals.(P＜0.05);.350 clinical transfusion records were also collected and 247 of which are considered valid.It appears there are numerous difference in Hb indexes among the tested hospitals and even the departments of internal medicine and surgical presented different takes on the subject as well.Conclusion The clinical blood transfusion management in Zhejiang province and the establishment of blood transfusion departments still need to be improved.A plan to manage clinical blood use need to be carried out ASAP,which would specify evaluation references,standardize personnel training and tech investments.Ultimately,we hope such actions would help to further regulate the clinical blood use in the region.

Objective To explore the influencing factors of late diagnosis for newly diagnosed HIV/AIDS positive patients in Guangxi in 2015.Methods The CD4 + T lymphocytes count which was first detection for newly diagnosed HIV/AIDS positive patients in Guangxi during 2015 was collected.Data were statistically analyzed.Results We collected 8 586 newly diagnosed HIV/AIDS whose median CD4+ T lymphocytes counts was 237.5 cells/μl,and 43.12％ of them had less than 200 cells/μl.Gender,age,occupation,marriage,nation,education,route of transmission,types of testing and region had effects on late HIV diagnosis(all P ＜ 0.05).Logistic analysis found that risk factors associated with the late diagnosis of HIV were male(OR =1.851,95％ CI:1.673-2.048),migrant worker (OR =1.387,95％ CI:1.242-1.549),education below middle and secondary school(OR =1.619,95％ CI:1.400-1.873),currently married(OR =1.207,95％ CI:1.075-1.354),divorced or widowed(OR =1.508,95％ CI:1.309-1.738).Voluntary testing was a protective factor.Conclusions The prevalence the late diagnosis of HIV was high in Guangxi in 2015,it is crucial for related departments to enhance the testing and screening effort for HIV/AIDS.

Objective To study the phylogenetic evolution and genetic variations of gag gene among the prevalent human immunodeficiency virus (HIV )‐1 strains in Guangxi Zhuang Autonomous Region . Methods Plasma samples of 158 HIV‐1 infected patients in Guangxi area were collected during October 2011 to March 2012 .The gag gene fragments of HIV‐1 were amplified by reverse transcription/nested‐polymerase chain reaction and then sequenced .MEGA 5 .03 was utilized to construct phylogenetic tree and to calculate the genetic distances and selection pressures (globle ω) of gag gene and its coding regions . The comparisons between two groups were tested by Student′s t test ,and the comparisons of multiple groups were tested by one‐way ANOVA .Results A total of 140 amplification products of gag gene were obtained from 158 samples .Four subtypes of HIV‐1 were found ,including CRF01_AE (80 ,57 .1% ) , CRF08_BC (46 ,32 .9% ) ,CRF07_BC (10 ,7 .1% ) ,and subtype B (B′) (4 ,2 .9% ) .The genetic distances of gag gene of the above subtypes were 0 .036 ± 0 .001 ,0 .031 ± 0 .002 ,0 .043 ± 0 .003 and 0 .102 ± 0 .006 ,respectively ,with statistical significance (F=220 .62 ,P<0 .01) .The p17 and p24 coding regions suffered negative selection pressure (globleω<1) .Neither the globle ω in p17 region nor that in p24 region had significant differences among different subtypes (F=0 .761 ,P=0 .469 and F=0 .037 ,P=0 .964 , respectively ) . Conclusion CRF01_AE is the major subtypes of HIV‐1 in Guangxi Zhuang Autonomous Region .The coding regions of gag gene are relatively conserved during evolution .Changes of HIV‐1 prevalence ,however ,may affect the genetic variation of gag gene ,which should be continuously monitored .

Objective To investigate the HIV-1 drug resistance in Guangxi during 2009 to 2012 and to analyze the correlations between drug resistance and HIV-1 subtypes.Methods Patients with human immunodeficiency virus infection or acquired immune deficiency syndrome ( HIV/AIDS) were randomly re-cruited from different areas in Guangxi.HIV-1 RNA was extracted from blood samples of the subjects and converted into complementary DNA ( cDNA) by using reverse transcription.The pol gene was amplified and sequenced.Subtyping analysis was performed by using the online analysis tool of Genotyping in combination with the MEGA 5.03 software.The HIV resistance mutations were determined and scored with the use of Stanford HIV Drug Resistance Database.Results A total of 196 pol gene sequences were obtained from 103 antiretroviral therapy (ART)-treated subjects (52.55%) and 93 ART-na?ve subjects (47.45%).The 196 pol gene sequences were classified into four subtypes including CRF01_AE, CRF08_BC, CRF07_BC and B, accounting for 48.47%, 44.90%, 6.12%and 0.51%, respectively.The HIV drug resistance rates in sub-jects with and without ART were 10.68% and 7.53%, respectively.Among the 196 subjects, 14 cases showed low level of drug resistance, 3 cases showed moderate level of drug resistance and 4 cases showed high level of resistance.Only one case was resistant to both nucleoside reverse transcriptase inhibitors ( NR-TIs) and non-nucleoside reverse transcriptase inhibitors ( NNRTIs) .The resistance rates of the 196 cases to protease inhibitor (PIs), NRTIs, NNRTIs, and integrase inhibitors (INs) were 6.63%, 3.06%, 11.22%and 8.67%, respectively.The frequencies of PIs-related mutations in subtypes CRF01_AE, CRF07_BC and CRF08_BC were 6.32%, 41.67% and 2.27%, respectively.Most of the PI-related A71V/T mutations were identified in strains belonging to subtype CRF07_BC, accounting for 75% of all A71V/T mutations found in the 196 strains.The NNRTI-related E138A mutations only appeared in strains belonging to subtype CRF08_BC.Conclusion The drug resistance rate among patients with HIV-1/AIDS in Guangxi was higher than the average level in China.The drug resistance rates varied with the subtypes of HIV-1 strains.

Objective To compare the genetic barriers to development of primary mutations related to drug resistance to protease inhibitors (PI), nucleioside reverse transcriptase inhibitors ( NRTI ), and non-nucleioside reverse transcriptase inhibitors ( NNRTI ) among human immunodeficiency virus (HIV)-1 CRF01_AE, CRF07_BC, and CRF08_BC strains, and to understand the difference of varying patterns of drug resistance related mutations within these subtypes. Methods One hundred and ninety naive HIV-positive subjects from Nanning City and Liuzhou City, Guangxi Zhuang Autonomous Region, were recruited. Peripheral blood samples were collected from all participants. HIV-1 RNAs were extracted from plasma, and the pol regions were amplified and sequenced. Sequences were subjected to phylogenetic analysis to determine the subtypes of HIV-1 isolates. Nucleotide transitions and transversions were counted for each primary mutation in these sequences. According to the phenomena that transitions occur on average 2. 5 times frequently than transversions, each transition was scored as 1, and each transversion scored as 2. 5. The sum of the scores for a particular substitution was calculated, and this value was taken as the genetic barrier to development of this mutation. Then, the differences of genetic barriers among the subtypes were assessed by Kruskal-Wallis test and Nemenyi test. Results A total of 123 sequences of CRF01_AE,CRF07_BC and CRF08_BC strains were selected. CRF08_BC had a lower genetic barrier for T/S69Dsubstitution than CRF01_AE and CRF07_BC (χ2 =107. 501, P＜0.01), while CRF01_AE and CRF07_BC had lower genetic barriers for V118I and L210W substitution than CRF08_BC. In addition,CRF07_BC had a decreased genetic barrier for V106M compared with CRF01_AE and CRF08_BC.Conclusions In the presence of the same selective pressure, subtypes CRF01_AE and CRF07_BC may be more likely to develop V118I and L210W substitution than CRF08_BC. However, CRF08_BC may be more likely to develop T/S69D substitution than CRF01_AE and CRF07_BC. Meanwhile, CRF07_BC may be easier to develop V106M substitution than CRF01_AE and CRF08_BC.

Objective To investigate the impact of methamphetamine (Meth) on the expressions of macrophage inflammatory protein (MIP)-1α ,MIP-1β ,interleukin (IL)-6 among human immunodeficiency virus(HIV)-infected patients .Methods The investigation was performed among 15 Meth-abuse and HIV-infected subjects (Meth + HIV ) ,15 non-Meth-abuse and HIV-infected subjects (non-Meth + HIV ) ,15 Meth-abuse and HIV-uninfected subjects (Meth) ,and 15 healthy subjects (HC) .CD4 + T lymphocyte counts in peripheral blood were detected by flow cytometry .The HIV viral loads in HIV-infected patients were detected by standard detection method .The levels of plasma MIP-1α ,MIP-1β and IL-6 from four groups were determined by enzyme-linked immunosorbent assay (ELISA ) .Intergroup difference was compared using t-test and interactive analysis was conducted using analysis of variance .Results In HIV-infected patients ,CD4 + T lymphocyte counts in Meth + HIV group was significant lower than non-Meth +HIV group (t= 5 .431 , P< 0 .01) ,whereas HIV viral load in Meth + HIV group was significant higher than non-Meth + HIV group (t= 4 .670 , P < 0 .01) .The levels of MIP-1α ,MIP-1β and IL-6 in Meth +HIV group were (40 .60 ± 9 .84) pg/mL , (47 .35 ± 11 .25 ) pg/mL and (37 .94 ± 11 .44 ) pg/mL , respectively ,and those in non-Meth + HIV group were (31 .31 ± 8 .11) pg/mL ,(39 .40 ± 8 .41) pg/mL and (31 .31 ± 8 .11) pg/mL ,respectively .The levels of MIP-1α ,MIP-1β and IL-6 in Meth + HIV group were all significantly higher than those in non-Meth + HIV group(t = 2 .822 , P= 0 .001 ;t = 2 .192 , P=0 .020 ;t= 1 .831 , P = 0 .043 ,respectively ) .The levels of MIP-1α ,MIP-1β and IL-6 in Meth group were (24 .45 ± 5 .90) pg/mL ,(27 .82 ± 7 .25) pg/mL and (27 .18 ± 8 .57) pg/mL ,respectively ,and those in HC group were (28 .42 ± 5 .79) pg/mL ,(31 .76 ± 9 .04) pg/mL and (23 .28 ± 6 .07) pg/mL ,respectively .But there were no significant differences of the levels of MIP-1α ,MIP-1β and IL-6 between Meth group and HC group(t= 1 .860 , P = 0 .158 ; t = 1 .317 , P = 0 .233 ; t = 1 .438 , P = 0 .228 ,respectively) .There was no association between Meth-abuse and the levels of these cytokines (P> 0 .05) ,neither between HIV infection and the levels of cytokines (P> 0 .05) .Conclusion Meth abuse results in elevated expressions of MIP-1αand MIP-1β ,which indicates that Meth abuse may play a regulating role on promoting HIV infection .

Objective To explore the expressions of Toll-like receptor 2 (TLR2 ) and the downstream proteins in patients with human immunodeficiency virus /Mycobacterium tuberculosis (HIV /M TB) co-infection .Methods A total of 119 subjects were randomly enrolled .The subjects were divided into four groups :HIV group (n = 32) ,HIV /M TB group (n = 30) ,M TB group (n = 28) and healthy control group (n= 29) .Peripheral venous blood was collected and the HIV-1 viral load was determined by standard method .The expression levels of TLR2 mRNA in peripheral blood mononuclear cells (PBMC) were determined by real-time quantitative PCR (qPCR) and mean fluorescent intensity (MFI) of TLR2 protein was detected by flow cytometry .The plasma interleukin (IL)-6 and tumor necrosis factor (TNF)-α levels were measured with enzyme-linked immunosorbent assay kits .The data were statistically analyzed by chi-square test ,students t test ,analysis of variance and rank sum test when appropriate .Results The viral load in HIV /M TB group ([5 .113 ± 1 .018] lg copy/mL ) was significantly higher than that in HIV group ([4 .416 ± 1 .020] lg copy/mL ; t = 3 .449 , P< 0 .01) .The TLR2 mRNA expressions in PBMC
among HIV ,HIV/M TB ,M TB and healthy control groups were 1 .397 ± 0 .601 ,1 .463 ± 0 .702 ,1 .429 ± 0 .630 ,and 0 .970 ± 0 .488 ,respectively ,which was significantly different among the 4 groups (F =4 .197 , P= 0 .007) .The MFI of TLR2 protein expressions on PBMC among HIV ,HIV /M TB ,M TB and healthy control groups were 28 .12 ± 4 .55 ,38 .11 ± 11 .77 ,31 .13 ± 12 .10 and 23 .33 ± 5 .14 ,respectively . The TLR2 protein expression levels were significantly different among 4 groups (F= 13 .976 ,P< 0 .01) . The plasma IL-6 and TNF-α concentrations were significantly different among 4 groups (Z = 19 .088 , 15 .475 ,both P< 0 .01) .The IL-6 concentrations in three patient groups were higher than that in healthy control group ,but the TNF-α concentrations were lower than healthy control group .Conclusions The co-infection of HIV-1 and M TB may enhance the activation of TLR2 signaling pathway ,which leads to the increased expression of IL-6 .

Objective To investigate the expression of TLR 4 and its downstream factor TNF-αin the patients with human immunodeficiency virus and Mycobacterium tuberculosis ( HIV/MTB) co-infection. Methods A total of 119 subjects including 32 patients with HIV infection (HIV group), 30 patients with HIV/MTB co-infection (HIV/MTB group), 28 patients with MTB infection (MTB group) and 29 healthy subjects ( control group ) were recruited continuously from the Fourth People′s Hospital of Nanning City , Guangxi.The expression of TLR4 in peripheral blood mononuclear cells (PBMCs) from the patients was de-termined by flow cytometry .ELISA was performed to detect TNF-αin plasma samples .The HIV-1 viral load was determined by standard method .Results The mean fluorescence intensity ( MFI) for TLR4 expression in PBMCs from HIV, HIV/MTB, MTB and control groups were 21.62±4.67, 18.29±3.87, 16.79±4.45, and 22.85±5.80, respectively, showing significant differences among four groups (F=8.105, P<0.01). The TLR4 levels in MTB and HIV/MTB groups were significantly lower than those in control group ( both P<0.01) and HIV group (P<0.01, P=0.014).The plasma concentrations of TNF-αin HIV, HIV/MTB, MTB and control groups were 15.892 (10.494-21.646) pg/ml, 13.142 (8.014-22.038) pg/ml, 16.284 (11.916-24.005) pg/ml, and 26.657 (16.321-34.541) pg/ml, respectively, that were significantly dif-ferent from each other (F=4.350, P=0.006).The levels of TNF-αin plasma from patients with HIV and HIV/MTB infection were significantly lower than those of healthy subjects (P=0.009 and P=0.001).The viral load in patients from HIV/MTB group (5.113 ±1.018 copies/ml) was significantly higher than that from HIV group (4.416±1.020 copies/ml) (t=3.449, P=0.001).Conclusion MTB infection might promote HIV replication by inhibiting the expression of TLR 4.HIV infection might increase host′s suscepti-bility to MTB infection by reducing the production of TNF-α.Suppressed expression of TLR and TNF-αpro-duction could contribute to the occurrence of HIV /MTB co-infection .

OBJECTIVETo investigate the distribution and proportion of subtypes of pol gene in HIV-1 epidemic strains in Guangxi Autonomous Region.

METHODS152 HIV-1 patients were enrolled from 11 cities in Guangxi Autonomous Region from 2010 to 2012 by convenient sampling. Inclusion criterias were listed as the fdlowing: HIV-1 infection was confirmed by Western blot, HIV-1 viral load >1 000 copies/ml, > 18 year-old, and without any serious illnesses. 5 ml of peripheral blood samples were obtained from each patient. The viral RNA was isolated from plasma and used for amplification of full-length pol gene by nested RT-PCR. The amplified products were sequenced. After editing and modification, all sequences were characterized for preliminary subtyping by genotyping and confirmed with phylogenetic tree constructed by MEGA 5.03 software. The recombinant identification of 2 unknown recombinant strains was determined by RIP and jpHMM at GOBICS.

RESULTSAmong 152 patients, 137 full-length pol genes were successfully amplified and 127 HIV-1 subtypes were identified. The distribution and proportion of subtypes was summarized as the following 71 cases of CRF01_AE, accounting for 55.9% (71/127), 38 CRF08_BC, 29.9% (38/127), 13 CRF07_BC, 10.2% (13/127), and 3 B (B'), 2.4% (3/127), 2 unknown recombinant strains, 1.6% (2/127). In 11 cites of Guangxi Autonomous Region, subtype CRF01_AE was the dominant strain. Among heterosexual transmitted patients and drug abusers, the proportions of subtype CRF01_AE were 67.4% (58/86) and 34.1% (14/41), respectively. There was a significance different in the distribution of CRF01_AE in different routes of transmission (χ(2)=15.07, P<0.001). In age 21- 35, age 36- 60 and age>60 groups, the proportions of CRF01_AE was 43.6% (17/39), 57.6% (38/66), 77.3% (17/22), and CRF08_BC was 43.6% (17/39), 28.8% (19/66), 9.1% (2/22), respectively, the difference in proportions was significant(χ(2)=8.48, P= 0.014). The patterns of two unknown recombinant strains were found to be CRF01_AE/B (B') and CRF01_AE/C/B(B'), respectively.

CONCLUSIONCRF01_AE was the dominant HIV-1 subtype in Guangxi Autonomous Region from 2010 to 2012, with heterosexual transmission as its main spreading route. The two unknown recombinant strains in Guangxi Autonomous Region were reconstructed by subtype CRF01_AE and CRF_BC.

Blotting, Western ; China ; epidemiology ; Cities ; Drug Users ; Genes, pol ; Genotype ; HIV Infections ; epidemiology ; transmission ; virology ; HIV-1 ; genetics ; Humans ; Phylogeny ; Polymerase Chain Reaction ; RNA, Viral ; blood ; pol Gene Products, Human Immunodeficiency Virus ; genetics

Objective:To study the intervention effect of ganoderma triterpenoids combined with exogenous monosialotetrahexosyl ganglioside(GM1) on cognitive dysfunction and synaptic ultrastructure of hippocampal neurons in rats with epilepsy caused by pentylenetetrazol(PTZ).Methods:A total of 40 Sprague-Dawley rats were divided into blank control group, epileptic model group, ganoderma triterpenoids group, GM1 group and GM1 combined with ganoderma triterpenoids group according to the random number table method( n=8 in each group). The rats were intraperitoneally injected with PTZ subconvulsant dose (35 mg·kg -1·d -1) once a day for 28 days to replicate the models of chronic epilepsy. And the rats in different medication groups were given corresponding administration based on daily intraperitoneal injection of PTZ(GM1: intraperitoneal injection of 30 mg·kg -1·d -1, ganoderma triterpenoids: gavage 1 000 mg·kg -1·d -1). Morris water maze was used to test the spatial exploration and learning and memory ability of epileptic rats.Transmission electron microscopy was used to observe the ultrastructure of hippocampal neurons in epileptic rats.Immunofluorescence staining was used to observe expression levels of cofilin and SYN protein in hippocampus CA1 of rats. In addition, Western blot was used to detect the expression levels of cofilin, p-cofilin and synaptophysin(SYN) protein in hippocampus of rats. SPSS 17.0 software was used for statistical analysis. Repeated one-way ANOVA was used for comparing among groups, LSD test was used for pairwise comparisons. Results:Morris water maze results showed that there were statistically significant differences in escape latency, times of crossing the platform and time spent in the target quadrant among the groups( F=5.259, 8.240, 5.961, all P<0.05). Compared with the epilepsy model group, the escape latencies((20.31±7.39) s, (21.81±6.05) s, (17.66±4.76) s) of the ganoderma triterpenoids group, GM1 group and GM1 combined with ganoderma triterpenoids group were shorter (all P<0.05), the numbers of crossing the platform ((4.63±1.41) times, (4.50±1.93) times, (5.50±1.77) times) were more (all P<0.05), the residence time in target quadrant ((31.91±5.00) s, (30.49±5.72) s, (35.70±5.34) s) were longer (all P<0.05). And the most obvious change was found in the GM1 combined with ganoderma triterpenoids group ( P<0.01). The results of transmission electron microscope showed that there were significant differences in the numbers of hippocampal neurons synapses, the synaptic gap, the density of postsynaptic membrane and length of active area of postsynaptic membrane among the groups( F=3.693, 7.201, 5.012, 4.033, all P<0.05). Compared with the epilepsy model group, the numbers of synapses ((8.00±1.79), (7.83±1.84), (8.50±1.87)) in the ganoderma triterpenoids group, GM1 group and GM1 combined with ganoderma triterpenoids group were all more (all P<0.05), synaptic gap ((33.83±3.81)nm, (32.43±4.14)nm, (30.23±3.08)nm)were narrower, and the postsynaptic dense substances ((57.50±6.03)nm, (58.10±2.40)nm, (60.73±3.81)nm) were all thicker (all P<0.05). The length of active region of postsynaptic membrane ((271.66±11.80) nm, (279.06±13.58) nm) in ganoderma triterpenoid group and GM1 combined with ganoderma triterpenoids group were longer than that in epilepsy model group (both P<0.05). Immunofluorescence results showed that the average fluorescence intensity of cofilin in the epilepsy model group was higher than that in the blank control group, and the average fluorescence intensity of SYN was lower than that in the blank control group (both P<0.05). The average fluorescence intensity of cofilin in GM1 group and GM1 combined with ganoderma triterpenoids group were lower than that in epilepsy model group (both P<0.05), and the average fluorescence intensity of SYN in ganoderma lucidum triterpenoids combined with GM1 group was higher than that in epilepsy model group ( P<0.05). Western blot showed that the expression levels of cofilin protein in the epilepsy model group was higher than that in the blank control group ((1.454±0.080), (1.092±0.099), P<0.05), and the expression of p-cofilin and SYN were lower than those in the blank control group ((1.103±0.120) vs (1.420±0.934), (1.650±0.062) vs (1.958±0.062), both P<0.05). The expression of cofilin protein ((1.227±0.071), (1.262±0.078), (1.162±0.129), P<0.05) in ganoderma triterpenoids group, GM1 group and GM1 combined with ganoderma triterpenoids group were lower than that in epilepsy model group, and the expression levels of p-cofilin(1.357±0.199) and SYN protein(1.873±0.010) in ganoderma triterpenoids combined with GM1 group were higher than that in epilepsy model group (both P<0.05). Compared with ganoderma lucidum triterpenoids group and GM1 group, there was no significant difference in each index of GM1 combined with ganoderma triterpenoids group (all P>0.05). Conclusion:GM1 combined with ganoderma triterpenoids may promote the synaptic plasticity of neurons, improve the learning and memory ability of epileptic rats.Combination medication is better than single medication in some observed indicators.