ObjectiveTo screen mimetic HIV-1 neutralizing epitopes from plasma with high level neutralizing antibody,and to provide useful information for further study of the interaction between antigen and antibody.MethodsIn order to gain neutralizing antibody recognized mimotopes, we detected neutralizing antibodieslevelsof 11HIV-1infectedslowprogressorsbyPBMC-basedneutralization assays.High-titer HIV-neutralizing antibodies from plasma of SPs was used as the ligand for biopanning by phage-displayed random peptide library.Positive phage clones was evaluated by ELISA,sequenced,and analyzed for homology to HIV-1 env by local BLAST to deduce the neutralizing peptide.ResultsTwenty-two clones were obtained consistent with requirement through three rounds biopanning.After comparison analysis,twelve clones include C8 were obtained as mimotopes of neutralizing antibody,C40 located in gp41Ⅱ cluster with the highest titer by inhibition ratio may be as neutralizing epitope.Conclusion By the use of IgG antibodies from SPs to screen the phage random polypeptide library,one can acquire multiple phage mimetic peptides of HIV related antigen epitope.(Chin J Lab Med,2012,35:838-842 )

Objective To build the cohort of drug resistance and analyze treatment efficiency of AIDS patients and situation of drug resistant mutations among HIV-1 infected individuals.Methods A cohort of 116 HIV-1 infected patients was built and their treatment progress were acquired once every 6 months.At the sanle time CD4+ T cell counts and HIV-1 viral load were measured and genotyping for drug resistance was determined by a home brew nested PCR.Results The CD4+ T cell count(470±251/ml)was higher than that before treatment in patients who were treated by AZT/DDI/NVP or D4T/DDL/NVP.The viral load was lower than that before treatmenL The drug resistant mutation frequency increased gradually along with treatment.The CD4+ T cell count was decreased and viral load was increased and the prevalence of drug resistant mutation was increased in the patients who changed regimens to AZT/3TC/NVP or D41/3TC/NVP.Only one primary mutation that was resistant to non-nucleoside reverse transcriptase inhibitors (NNRTIs)was detected in the naive patients.The cross-resistant mutation was detected in two patients after 6 months treatment. The intermediate resistance to lopinavir(LPV) was detected after 12 months treatment.The prevalence of high-grade resistances to NNRTIs was increased obviously,and the prevalence of multi-resistance and cross-resistance was detected in 5 patients after 36 months treatment.Conclusions The prevalence of primary mutation was rare in naive HIV-1 infected patients.The prevalence of drug resistant mutation was inereased gradually along with treatment.Ahhough few regimens were available,the treatment effect could last relatively long period of time if patients keep taking medicine stably.The regimens could be changed according to the results of drug resistant test.

Objective To investigate the efficacy and safety of autologous cryopreserved platelet transfusion in the management of thrombocytopenia after chemotherapy in hematological malignancy.Methods A total of 40 patients diagnosed as hematological malignancy with complete remission were equally assigned into study group and control group.During chemotherapy interval in the study group,when platelet counts exceeded 120 × 109/L,autologous platelets were collected with CS3000 Cell Separator and cryopreserved at-80℃ with 5％ dimethylsulfoxide.When platelet counts dropped below 15 × 109/L after chemotherapy,autologous platelets were thawed with 40℃ water bath and transfused back to each patient.In the control group,when platelet counts dropped below 15 × 109/L after chemotherapy,allogeneic fresh platelets were transfused.Median loss during the freeze-thaw-wash procedure in study group was observed,and the 1 h,24 h corrected count increments(CCI)were calculated in the both groups.The hemostatic effects and adverse reactions were also observed.Results In the control group,1hCCI and 24hCCI were (19.3 ±6.1)× 109/L and(12.2 ± 7.0)× 109/L,respectively,with the effective rate of 80％ and the transfusion reaction rate of 45％.Totally 20 collection and transfusions were finished in the study group.A total of(3.4-8.5)× 1011 platelet were obtained in each collection.Platelet recovery after freezing and thawing was(73.51 ±9.03)％(62％-83％).1hCCI was(17.4±7.6)× 109/L,24h CCI was(10.5 ±5.8)× 109/L and the effective rate was 85％.There was no significant different between the two groups (P ＞ 0.05).The transfusion reaction rate was 15 ％,which was significantly lower than that of the control group(P ＜ 0.05).Meanwhile,adverse reactions were occurred less in the study group.Conclusion This study demonstrates that autologous cryopreserved platelet transfusions can be safely administered for supporting thrombocytopenia in hematological malignancy patients undergoing chemotherapy.

Objective To investigate the accuracy of serum S-100β protein and neuron specific enolase (NSE) level in predication of postoperative delirium (PD) in patients of different ages.Methods Four hundred ASA Ⅰ -Ⅳ patients of both sexes weighing 40-82 kg undergoing abdominal surgery performed under general anesthesia were divided into 4 age groups:group Ⅰ 18-44 yr; group Ⅱ 45-59 yr; group Ⅲ 60-74 yr and group Ⅳ ≥75 yr.The diagnosis of PD was made by using confusion assessment method.The incidence of PD was recorded within 72 h after operation.Each group was further divided into PD and non-PD subgroups.Blood samples were taken at 1 day before operation (T1),during their stay in PACU (T2) and at 24 and 72 h after operation (T3,4 ) for determination of serum S-100β protein and NSE concentrations.The receiver operating characteristic (ROC) curve for serum S-100β protein concentration in determining the PD efficacy was plotted.Results The incidence of PD was significantly higher and the duration was significantly longer in groups Ⅲ and Ⅳ than in groups Ⅰ and Ⅱ,and in group Ⅳ than in group Ⅲ (P ＜ 0.05).There was no significant difference in the serum S-100β protein concentration between PD subgroup and non-PD subgroup in groups Ⅰ - Ⅲ ( P ＞ 0.05).Compared with that at T1 and in nonPD subgroup,the serum S-100β protein concentration was significantly increased in PD subgroup in group Ⅳ,and the serum NSE concentration was significantly decreased at T2,3 in PD subgroup in group Ⅰ (P ＜ 0.05).There was no significant difference in the serum NSE concentration between PD subgroup and non-PD subgroup in groups Ⅱ -Ⅳ.The analysis results of the ROC curve showed that:the area under the curve for the serum S-100β protein concentration and 95％ confidence interval were 0.329 (0.127-0.531),0.352 (0.168-0.536),0.619 (0.466- 0.772) and 0.921 (0.846-0.995),the sensitivity was 50％,50％,56％ and 88％,and the specificity was 29％,22％,46％ and 86％ in groups Ⅰ-Ⅳ respectively.Conclusion Increase in the serum S-100β protein concentration can be used in predicting the development of PD in patients ≥75 yr,but the serum NSE protein concentration can not be used.

Objective We aim to evaluate the value of the current serum creatinine reference interval ( RI ) provided by Industry Standard WS /T 404.5 in clinical practice.Methods The first time serum creatinine levels and urinary albumin /creatinine ratio were obtained from 67 605 adult patients ( <60 years old) who were treated in the First Hospital of China Medical University between October 1, 2014 and September 30, 2015 in this cross-sectional study.Estimated glomerular filtration rate ( eGFR ) calculated by chronic kidney disease epidemiology collaboration (CKD-EPI) equation and urinary albumin /creatinine ratio (ACR) were used to evaluate the clinical practical significance of current and old serum creatinine RIs as the criteria.Results 4.3% of normal subjects based on current RI were showed decreased eGFR, 98% of abnormal subjects based on the current RI were founded to have decreased eGFR . 1378 subjects were evaluated as increased based on current RI but as normal based on old RI , and 93.5% of these subjects were showed decreased eGFR .In addition, ACR was measured in 26 cases, and 18 out of 26 cases (69.2%) were confirmed to have elevated ACR (≥30 mg/g) and proteinuria.On the other hand of analysis, screening positive rates of declined eGFR were 43.6% by old RI and 61.9% by current RI in the subjects with eGFR under 90 ml(min ×1.73 m 2 ), and the performance of the current RI was obviously improved(χ2 =212.648,P <0.001).Conclusions The current reference interval of serum creatinine is favorable for the detection of renal dysfunction in patients .It is recommended that the current reference interval can be applied in the clinical laboratories as early as possible .

OBJECTIVETo determine if tanshinone IIA (Tan IIA) can influence the development of elastase-induced experimental abdominal aortic aneurysms (AAAs).

METHODSTotally 36 male Sprague-Dawley rats were randomly distributed into three groups (12 in each group): Tan IIA group, control group, and sham group. Rats of Tan IIA and control groups underwent intra-aortic elastase perfusion to induce AAAs, while rats of sham-group were perfused with saline. Rats of Tan IIA-group received Tan IIA treatment (2 mg/d). The luminal diameter of the aneurysm at the segment with maximum diameter were measured pre-perfusion and on the 4 time point after perfusion. Systolic blood pressure was measured by tail-cuff technique. Aortic tissue samples were obtained on 24 days after perfusion and evaluated by RT-PCR, Western blot, immunohistochemistry and Miller's elastin-Van Gieson's (EVG) staining.

RESULTSTwenty-four days after perfusion, Tan IIA significantly reduced increased aortic size compared to control group ((0.210 ± 0.002) cm vs. (0.304 ± 0.004) cm, t = 78.858, P = 0.000) without affecting blood pressure (t = -1.237 to -1.221, P > 0.05). The over-expression of matrix metalloproteinases (MMP)-2/9 (t = 25.943, P = 0.000; t = 42.815, P = 0.000), monocyte chemotactic protein-1 (MCP-1) (t = 4.518, P = 0.000), inducible nitric oxide synthase (iNOS) (t = 17.685, P = 0.000) and the destruction of elastic fibers in aortic tissue of control group were significantly less than Tan IIA group (0.469 ± 0.040 vs. 0.230 ± 0.024, t = 17.944, P = 0.000).

CONCLUSIONTanshinone IIA attenuates the development of elastase-induced experimental AAAs possibly by down-regulating MMP-2/9, MCP-1 and iNOS expression.

Animals ; Aortic Aneurysm, Abdominal ; chemically induced ; drug therapy ; Chemokine CCL2 ; metabolism ; Disease Models, Animal ; Diterpenes, Abietane ; therapeutic use ; Elastic Tissue ; metabolism ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Pancreatic Elastase ; adverse effects ; Rats ; Rats, Sprague-Dawley

Objective To observe the effects of Najia method of midday-midnight point selection for acute ischemic stroke (AIS) model rats onthe contents of NSE and S100B protein in serum. Methods SPF SD male rats were chosen to establish the models by middle cerebral artery bolt method. Rats were divided into blank group, sham-operation group, model group, channel-point group, and Najia method group by random number table method. Blank group, sham-operation group, and model group were in the absence of treatment, while the channel-point group received acupuncture treatment according to differentiation syndrome. Najia method group used Najia method of midday-midnight point selection to conduct acupuncture treatment once a day. Improvement of neural function and cerebral infarction volume were observed. The contents of NSE and S100B protein in serum were detected. Results Compared with model group, neurological function score, infarct volume and infarct volume percentage, and the contents of NSE and S100B protein in serum decreased in Najia method group and channel-point group (P<0.05, P<0.01). The effects of Najia group were generally better than the channel-point group. Conclusion Najia method of midday-midnight point selection can decrease the content of NSE and S100B protein in serum of AIS model rats, so as to achieve the effects of neuroprotection and treatment.

Objective To examine the expression of TLR7/8 in monocytes purified from HIV-1 infected individuals and to study its association with disease progression. Methods Sixty-three HIV-1 infected individuals and 18 normal controls were enrolled. Monocytes were purified by MACS system and RNA of them was extracted by RNA mini kit of QIAGEN company. TLR7/8 expression was tested by real-time RT-PCR with ABI7500. Results It was found that the expression of TLR7 was strongly correlated with absolute CD4 count (r =0.614, P＜0.01) , so was TLR8 (r =0.419, P＜0.01). The expression of TLR7 in slow progressor (SP) group was higher than that in HIV-1 infected patients group, AIDS patients group and normal group (P ＜ 0. 05 ) . HIV group and normal group were strongly higher than AIDS group (P ＜ 0. 05). It was no significant differentiation of expression of TLR7 between HIV infection group and normal control group. The expression of TLR8 in SP group and normal group were significantly higher than that in AIDS group (P ＜ 0. 05). The expression of TLR8 was no singnificantly difference between SP group and HIV group or normal control group, so was it between HIV group or normal control group and AIDS group. Conclusion The expression of TLR7/8 in monocytes from HIV-1 infected patients significantly correlated with disease progression.

Objective To investigate the killer cell lg-like receptors (KIR) gene frequency of HIV-1 infected slow progressors(SP) and typical progressors(TP), and to analyze the interaction between KIR alleles and the progression of HIV-1 infection in Chinese population. Methods Eighty-one HIV-1 posi-tive individuals including 43 SPs and 38 TPs were recruited. Carriage of KIR genes was assessed using poly-merase chain reaction sequence-specific primers (PCR-SSP) assays. Results KIR2DS3 gene frequency was significantly lower in SP group (3.6%) than that in TP group (14.2%), P =0. 018 ,OR =0. 210,95% CI =0.053-0.833. The number of activating KIR genes was less in SP group than that in TP group, but was not significant (P = 0. 208). Conclusion Lower KIR2DS3 gene frequency may potentially be associated with slower progression to AIDS in Chinese population.

Objective To explore the effects of mesenchymal stem cells ( MSC ) treatment on platelet counts in mice with immune-mediated thrombocytopenia ( ITP) and the possible mechanism .Meth-ods ITP was induced by daily intraperitoneal injection of anti-platelet membrane CD 41 antibody (MWReg30) into BALB/c mice.The mice were then divided into experiment and control groups with 20 mice in each.Each mouse in experimental group was injected with 2×107 mesenchymal stem cells (MSC) through the tail vein .The numbers of blood platelets in mice from two groups were counted on days 5, 7 and 14 after MSC injection .Reverse transcriptase polymerase chain reaction ( RT-PCR) was performed to meas-ure T-bet and GATA-3 gene expression in peripheral blood mononuclear cells ( PBMCs ) at mRNA level on day 14.The levels of IFN-γ, IL-2, IL-4 and IL-10 in serum were detected by ELISA .Results The platelet counts in experimental group were significantly higher than those in control group on days 7 and 14 after MSC injection [(588.0±81.6)×109/L and (623.0±78.9) ×109/L vs.(317.0±90.1) ×109/L and (288.0± 87.8)×109/L ] (P<0.05).On day 14 after MSC injection, the T-bet expression at mRNA level in PBMCs from mice in experimental group was significantly lower than that in control group [(0.04±0.03) vs.(0.27 ±0.05)] (P<0.05), while the GATA-3 expression at mRNA level was higher than those in control group [ (0.14±0.04) vs.(0.07±0.05)] (P<0.05).Compared with control group, the concentrations of Th1 type cytokines such as IFN-γand IL-2 were remarkably down-regulated in experimental group [(3.1±1.7) pg/ml and (3.2±2.1) pg/ml vs.(10.3±4.8) pg/ml and (16.3±5.7) pg/ml](P<0.05), while the con-centrations of Th2 type cytokines such as IL-4 and IL-10 were up-regulated in experimental group [(88.6± 15.2) pg/ml and (38.3±11.8) pg/ml vs.(32.7±5.7) pg/ml and (22.1±3.4) pg/ml ] (P<0.05). Conclusion MSC treatment can effectively increase platelet counts in mice with immune-mediated thrombo-cytopenia, which may be associated with the suppression of Th 1-dominant response mediated by abnormal ex-pression of T-bet and GATA-3.