The application value of Multi-Latex polygranular technique joint detection of urinary microproteins in noninvasive diagnosis after renal transplantation / 器官移植

Objective To investigate the application value of Multi-Latex polygranular technique joint detection of kidney injury-related urinary microproteins in noninvasive diagnosis after renal transplantation. Methods Clinical data of 72 recipients undergoing renal transplantation were retrospectively analyzed. According to the level of serum creatinine (Scr), the recipients were divided into normal renal function group (group A, n=14), mild kidney injury (group B, n=37), and severe kidney injury group (group C, n=21). 20 healthy volunteers were selected as the healthy control group (HC group). The contents of urinary retinol binding protein (RBP), microalbumin (mAlb), IgG, transferrin (TRF), α1-microglobulin (MG), and β2-MG of subjects in each group were detected using the Multi-Latex polygranular technique. The correlation between urinary microproteins and Scr, blood urea nitrogen (BUN) was analyzed. The differences of urinary microproteins in each group were compared. And the diagnostic value of single and joint detection of urinary microproteins was evaluated. Results Six kinds of urinary microproteins in HC group and group A were significantly lower than those in group B and group C, and six kinds of urinary microproteins in group B were significantly lower than those in group C (all P < 0.01). Six kinds of urinary microproteins in renal transplant recipients were positively correlated with BUN. RBP, mAlb, α1-MG, and β2-MG were positively correlated with Scr. The correlations were statistically significant (P < 0.001-0.05). The diagnostic value of joint detection of urinary microproteins is better than the detection of single index, among which TRF+mAlb+RBP+α1-MG quadruple detection had the highest diagnostic value. Conclusions Six kinds of urinary microproteins can be used as specific indicators to reflect graft renal function. The polygranular technique can simultaneously detect its contents and achieve noninvasive diagnosis. The diagnosis based on TRF+mAlb+RBP+α1-MG quadruple detection is expected to further improve the noninvasive diagnosis system after renal transplantation.

Effect of oral motor intervention on oral feeding ability and growth of neonates with intestinal atresia / 中国实用护理杂志

Objective:To explore the effect of oral motor intervention on oral feeding ability and growth of neonates with intestinal atresia.Methods:A total of 80 intestinal atresia neonates in the Hunan Children`s Hospital from January 2017 to January 2019 were admitted to the present study. Neonates were randomly assigned to oral motor group and control group according to the random number table. The control group received routine nursing, while the oral motor group carried out oral motor intervention for 14 consecutive days. Both groups were followed up for six months. The oral feeding ability and growth index such as body weight, body length and head circumference were compared between two groups.Results:The complete oral feeding rate transfer, complete oral feeding rate transfer, time to complete oral feeding were (10.48±2.96) ml/min, (90.02±8.36) %, (15.06±2.99) days in the oral motor group, those index were (8.18±2.44) ml/min, (72.58±9.46) %, (18.08±4.98) days in the control group, the differences were statistically significant ( t values were 3.633, 8.316, 3.106, P<0.01). After 7, 14 days of intervention, the non-nutritive sucking scores were (52.24±8.89) points, (69.81±12.94) points in the oral motor group, significantly higher than (48.08±6.72) points, (63.09±11.73) points in the control group, the differences were statistically significant ( t values were 2.265, 2.327, P<0.05). After 3, 6 months of intervention, the levels of body weight were (6 234.21±560.25) g, (7 630.19±782.25) g, significantly higher than (5 916.89±462.40) g, (7 293.65±979.98) g in the control group, the differences were statistically significant ( t values were 2.648, 2.148, P<0.05). After 6 months of intervention, the levels of head circumference were (43.81±5.59) cm in the oral motor group, significantly higher than (40.85±3.73) cm in the control group, the difference was statistically significant ( t value was 2.635, P<0.05). Conclusion:Oral motor intervention can promote oral feeding ability and improve growth performance of neonates with intestinal atresia.

Intervention of family-integrated care model for children with enterostomy and their caregivers / 中国实用护理杂志

Objective To investigate the effects of family-integrated care (FIC) on postoperative outcomes in children with enterostomy and their caregivers. Methods From August 2017 to August 2018, 50 children with enterostomy and 50 family members of the Children′s Hospital of Hunan Province were selected as subjects. According to the random number table, the children and their families were divided into control group and the observation group, each group was 25 cases. The control group was given a routine nursing mode to intervene, and the observation group was given an FIC mode for intervention. Postoperative outcomes were evaluated using the incidence of ostomy complications and readmission rates. The pre-intervention and outpatient follow-up were used to assess the psychological status of the family members using the Self-rating Anxiety Scale (SAS) and the Self-rating Depression Scale (SDS), and to assess the postoperative care of the family's intestines using the postoperative evaluation of the postoperative intestines. The level of knowledge mastery. The self-rating anxiety scale (SAS) and the self-rating depression scale (SDS) were used to assess the psychological state of the family members, and the postoperative care knowledge evaluation form for the postpartum was used to evaluate the postoperative care of the family. Results The incidence of ostomy complications in the observation group was 8.70% (2/23), which was lower than that in the control group (34.78% (8/23) (P<0.05). The readmission rate of the observation group was 0(0/23), which was lower than the control group 17.39% (4/23) (P<0.05). After intervention, the SAS scores and SDS scores of the families of the two groups were lower than those before the intervention, and the SAS scores and SDS scores of the observation group were lower than the control group(P<0.05). Before discharge and 3months of follow- up, the scores of postoperative care knowledge evaluation scores of the observation group were higher than those of the control group(P<0.05). Conclusions FIC mode can effectively reduce the incidence of complications and readmission rate in children with enterostomy. It has positive significance for improving the negative emotions of children′s family members and improving the mastery of postoperative care.

Intervention of family-integrated care model for children with enterostomy and their caregivers / 中国实用护理杂志

Objective@#To investigate the effects of family-integrated care (FIC) on postoperative outcomes in children with enterostomy and their caregivers.@*Methods@#From August 2017 to August 2018, 50 children with enterostomy and 50 family members of the Children′s Hospital of Hunan Province were selected as subjects. According to the random number table, the children and their families were divided into control group and the observation group, each group was 25 cases. The control group was given a routine nursing mode to intervene, and the observation group was given an FIC mode for intervention. Postoperative outcomes were evaluated using the incidence of ostomy complications and readmission rates. The pre-intervention and outpatient follow-up were used to assess the psychological status of the family members using the Self-rating Anxiety Scale (SAS) and the Self-rating Depression Scale (SDS), and to assess the postoperative care of the family's intestines using the postoperative evaluation of the postoperative intestines. The level of knowledge mastery. The self-rating anxiety scale (SAS) and the self-rating depression scale (SDS) were used to assess the psychological state of the family members, and the postoperative care knowledge evaluation form for the postpartum was used to evaluate the postoperative care of the family.@*Results@#The incidence of ostomy complications in the observation group was 8.70% (2/23), which was lower than that in the control group (34.78% (8/23) (P<0.05). The readmission rate of the observation group was 0(0/23), which was lower than the control group 17.39% (4/23) (P<0.05). After intervention, the SAS scores and SDS scores of the families of the two groups were lower than those before the intervention, and the SAS scores and SDS scores of the observation group were lower than the control group(P<0.05). Before discharge and 3months of follow-up, the scores of postoperative care knowledge evaluation scores of the observation group were higher than those of the control group(P<0.05).@*Conclusions@#FIC mode can effectively reduce the incidence of complications and readmission rate in children with enterostomy. It has positive significance for improving the negative emotions of children′s family members and improving the mastery of postoperative care.

Activation of CD8+T cells regulated by γ-aminobutyric acid and its receptors / 器官移植

Objective To evaluate the effect of γ-aminobutyric acid (GABA) and its receptors upon the proliferation of CD8+T cells.Methods The splenic CD8+T cells of Balb/c mice were obtained by CD8+f cell magnetic bead sorting kit.Under the effect of CD3/CD28-activated magnetic bead,CD8+T cells were stimulated by different concentrations of GABA.5-bromo-2-deoxyuridine (BrdU) labeling and flow cytometry were performed to detect the proliferation of CD8+T cells.The expression levels of GABA-A and GABA-B receptor before and after CD8+T cell activation were compared by fluorescent quantitative real-time polymerase chain reaction (PCR).Result Flow cytometry result revealed that GABA could inhibit the proliferation of activated CD8+T cells,manifested as significant decrease in the quantity of CD152+CD8+T cells.Fluorescent quantitative real-time PCR demonstrated that GABA-A receptor subtypes α2,α6 and GABA-B receptor subtype 1a were expressed only when the CD8+T cells were activated.After CD8+T cell activation,the quantity of GABA-A receptor subtypes α3,αs,β2,β3,γ1,γ2 and θ were significantly increased,whereas the quantity of GABA-B2R and GABA-B1b did not significantly differ before and after CD8+T cell activation.Conclusions GABA can suppress the proliferation of activated CD8+T cells.The activation of CD8+T cells is regulated by GABA receptors.However,the underlying mechanism remains to be elucidated.

Simultaneous Determination of 6 Residual Organic Solvents in Xingnaojing Injection by Headspace GC / 中国药房

OBJECTIVE:To establish the method for simultaneous determination of 6 residual organic solvents in Xingnaojing injection,such as methanol,ethanol,isopropanol,n-butanol,ethyl acetate and acetonitrile.METHODS:Headspace GC method was adopted.The determination was performed on DB-624 capillary column by temperature programming with the injector temperature of 200 ℃;flame ionization detector was adopted with the temperature of 250 ℃;carrier gas was nitrogen with flow rate of 25 mL/min and split ratio of 35 ∶ 1;headspace sampling size was 1 mL,and heating temperature of headspace sampling was 80 ℃;equilibrium time was 15 min.RESULTS:The linear ranges of methanol,ethanol,isopropanol,n-butanol,ethyl acetate and acetonitrile were 15.00-240.00 μg/mL (r =0.999 9),25.00-400.00 μg/mL (r =0.999 9),25.00-400.00 μg/mL (r =0.999 9),25.00-399.99 μg/mL(r=0.999 9),25.00-399.99 μg/mL(r=0.999 8) and 5.00-80.00 μg/mL(r=0.999 9).The LOQ were 5.98,3.94,2.05,2.13,1.39,1.24 μg/mL,and the LOD were 2.01,2.11,1.18,1.56,1.15,0.01 μg/mL,respectively.RSDs of precision tests were all less than 2.0％,stability and repetitive tests only ethyl acetate was detected,RSD＜2.0％;the recoveries were 93.59％-99.02％ (RSD=2.62％,n=6),92.42％-98.40％ (RSD=2.43％,n=6),94.81％-104.64％ (RSD=3.47 ％,n=6),94.56％-106.73％ (RSD=4.21％,n=6),97.04％-106.33％(RSD=3.50％,n=6)and 98.40％-107.97％ (RSD=3.37％,n=6).CONCLUSIONS:The method is specific,rapid,simple and accurate,and can be used for simultaneous determination of 6 residual organic solvents in Xingnaojing injection.

Simultaneous Determination of 6 Residual Organic Solvents in Xingnaojing Injection by Headspace GC / 中国药房

OBJECTIVE:To establish the method for simultaneous determination of 6 residual organic solvents in Xingnaojing injection,such as methanol,ethanol,isopropanol,n-butanol,ethyl acetate and acetonitrile.METHODS:Headspace GC method was adopted.The determination was performed on DB-624 capillary column by temperature programming with the injector temperature of 200 ℃;flame ionization detector was adopted with the temperature of 250 ℃;carrier gas was nitrogen with flow rate of 25 mL/min and split ratio of 35 ∶ 1;headspace sampling size was 1 mL,and heating temperature of headspace sampling was 80 ℃;equilibrium time was 15 min.RESULTS:The linear ranges of methanol,ethanol,isopropanol,n-butanol,ethyl acetate and acetonitrile were 15.00-240.00 μg/mL (r =0.999 9),25.00-400.00 μg/mL (r =0.999 9),25.00-400.00 μg/mL (r =0.999 9),25.00-399.99 μg/mL(r=0.999 9),25.00-399.99 μg/mL(r=0.999 8) and 5.00-80.00 μg/mL(r=0.999 9).The LOQ were 5.98,3.94,2.05,2.13,1.39,1.24 μg/mL,and the LOD were 2.01,2.11,1.18,1.56,1.15,0.01 μg/mL,respectively.RSDs of precision tests were all less than 2.0％,stability and repetitive tests only ethyl acetate was detected,RSD＜2.0％;the recoveries were 93.59％-99.02％ (RSD=2.62％,n=6),92.42％-98.40％ (RSD=2.43％,n=6),94.81％-104.64％ (RSD=3.47 ％,n=6),94.56％-106.73％ (RSD=4.21％,n=6),97.04％-106.33％(RSD=3.50％,n=6)and 98.40％-107.97％ (RSD=3.37％,n=6).CONCLUSIONS:The method is specific,rapid,simple and accurate,and can be used for simultaneous determination of 6 residual organic solvents in Xingnaojing injection.

Content Determination of Paeoniflorin in Bazhen Pill(Concentrated Pill)by HPLC / 中国药房

OBJECTIVE:To establish a method for the content determination of paeoniflorin in Bazhen pill(concentrated pill). METHODS:HPLC was performed on the column of wondasil C18 with mobile phase of acetonitrile-1% phosphoric acid(15∶85, V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 230 nm,and column temperature was 30 ℃. RESULTS:The lin-ear range of paeoniflorin was 0219-1.32 μg(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 1%;recovery was 95.82%-101.82%(RSD=2.13%,n=6). CONCLUSIONS:The method is simple,stable and reproducible, and can be used for the content determination of Bazhen pill(concentrated pill).

Effects of MF59 in combination with heat-killed BCG as adjuvant on the immunogenicity of Mycobacteri-um tuberculosis fusion protein PstS1-LEP / 中华微生物学和免疫学杂志

Objective To study the effects of MF59 in combination with heat-killed BCG ( hBCG) as adjuvant on the immunogenicity of Mycobacterium tuberculosis fusion protein PstS1-LEP.Methods BALB/c mice were divided into six groups from group 1 through group 6.They were immunized with PstS1-LEP+MF59 ( group 1 ) , PstS1-LEP+MF59/hBCG ( group 2 ) , PstS1-LEP+hBCG ( group 3 ) , MF59 ( group 4 ) , PstS1-LEP (group 5) and hBCG (group 6) for three times at intervals of two weeks , respectively.The mice were sac-rificed two weeks after the last immunization .The serum samples were collected for antibodies detection .The splenic lymphocytes and peritoneal macrophages were isolated and cultured with PstS 1-LEP.Indirect ELISA and sandwich ELISA were used to detect PstS 1-LEP-specific antibodies and cytokines in the supernatants of culture , respectively.Results The level of IFN-γ, IL-1β, IgG, IgG1 and IgG2a in group 1 were higher than those in groups 4, 5 and 6 (P<0.05).The level of IL-2 and IL-4 in group 1 were higher than those in groups 4 and 6 (P<0.05).The level of IFN-γ, IL-1β, IL-12, IgG, IgG1 and IgG2a in group 2 were higher than those in groups 4, 5 and 6 (P<0.05).The level of IL-2 was higher in group 2 than that in groups 4 and 6 (P<0.05). The level of IL-4 in group 3 was higher than that in group 4 ( P=0.05 ) .The level of IL-1βin group 3 were higher than that in groups 4 and 5 ( P<0.05 ) .The level of IgG was higher in group 3 than that in groups 4 and 6 (P<0.05).IgG1 level in group C was up-regulated in comparison with that in groups 4, 5 and 6 (P<0.05 ) .Conclusion hBCG as PstS1-LEP adjuvant induces a shift towards Th 2-type immune response , while MF59 induces Th1/Th2-type immune response.The combination of MF59 and hBCG inhibits the secretion of IL-4 by spleen lymphocytes , but enhances the secretion of IL-12 by macrophage .

γ-aminobutyric acid B receptor (GABABR)ameliorated liver fibrosis by inhibiting hepatic cell migration / 中华肝胆外科杂志

Objective To investigate the role of r-aminobutyric acid B receptor in the development of liver fibrosis.Methods Thirty-two Sprague-Dawley (SD) rats were divided into four groups including a control group,a model group,a baclofen group,and a CGP35348 group.Liver fibrosis was then induced by carbon tetrachloride (CCl4).Baclofen and CGP35348 treatment were carried out after the formation of liver fibrosis,followed by complete extraction of the eyeball to obtain blood sample to test liver function.Liver tissue specimens were cut and stored for histological staining,histochemistry,real-time polymerase chain-reaction (RT-PCR),and western blot analysis.Results Histological staining indicated that the degree of liver fibrosis was more severe in the CGP35348 group than in the baclofen group (P＜0.001).The levels of alanine transaminase (ALT),aspartate aminotransferase (AST),gamma-glutamyl transferase (GGT),total bilirubin (TBil),and direct bilirubin (DBil) were significantly lower in the baclofen group than in the CGP35348 group (P＜0.01).The levels of ALT,AST,GGT,TBil,and DBil were significantly higher in the CGP35348 group than in the model group (P＜0.05).Immunofluorescence results show that the hepatic cell migration was inhibited in the baclofen group.Western blot results showed that the expression levels of α-SMA protein were significantly lowered in the baclofen group when compared to that of the CGP35348 group and model group (P＜0.01).Conclusion GABAB receptor might play a role in the liver protection by inhibition of migration of hepatic cells in liver fibrosis.Further studies into the mechanism behind this function are further needed and may be a potential source of future anti-fibrotic treatment.

Impacts of concentrations of Tacrolimus on NK cells and receptors in peripheral blood of renal transplant recipients / 中华器官移植杂志

Objective To study the effects of tacrolimus(Tac) concentrations on the number of NK cells and receptor expression in peripheral blood of renal transplantation receptors.Methods A total of 60 first-time kidney transplantation recipients in our institute from Dec.2007 to July 2009 were followed up.Tac maintenance immunosuppressive therapy was given to all recipients.The recipients were divided into low-concentration Tac group (6.84 + 1.72μg/L,n =30) and highconcentration Tac group ( 11.88 + 2.59 μg/L,n =30) according to concentrations of Tac.Twenty healthy volunteers served as controls.Before and 6 months after operation,concentrations of Tac were analyzed by using micro particle immunoassay chemiluminescent method.NK cells and their receptors (CD85j,CD158d,CD94 and NKG2D) were detected by using flow cytometry.The concentrations of soluble HLA-G5 were detected by using ELISA.Results The number of NK cells in lowconcentration Tac group and high-concentration of Tac group preoperatively was significantly reduced as compared with control group (P ＜ 0.05 ). The percentage and number of NK cells in low concentration Tac group and high-concentration Tac group at 6th month after operation were significantly reduced as compared with control group (P＜0.05).The number of NK cells in lowconcentration Tac group was significantly greater than in high-concentration Tac group (P＜ 0.05).There was no significant differende in the expression of CD85j,CD158,CD94 and NKG2D before operation between two groups(P＞0.05).The expression of CD85j and CD158d in two groups was increased,but that of CD94 and NKG2D was decreased at 6th month post-transplantation as comapred with that preoperation.In low-concentration Tac group,the expression of CD85j and CD158d was increased as compared with that in high-concentration Tac group (P＜0.05 ).Spearman correlation analysis revealed that the CD85j and CD158d expression had a positive correlation with sHLA-G5(P＜0.01 ),but the NKG2D had a negative correlation with sHLA-G5(P＜0.01 ).Conclusion There was correlation between the concentrations of Tac and NK cells count and NK receptors. Low concentrations of Tac can safely and effectively protect kidney function.The number of NK cells andtheir inhibitor receptors are increased in the recipients with low concentration of Tac.

Preparation of recombinant PPE65 protein of Mycobacterium tuberculosis and its applications in serodiagnosis of patients with pulmonary tuberculosis / 中华检验医学杂志

Objective To evaluate the potential value of IgG antibodies against recombinant PPE65 protein (rPPE65) of Mycobacterium tuberculosis in serodiagnosis of tuberculosis.Methods The gene encoding PPE65 protein of M.tuberculosis was cloned into the PET-28a vector and then expressed in Escherichia coli.The rPPE65 was purified with Ni-NTA affinity and ion exchange chromatography.After dialysis renaturation, the concentration of rPPE65 was determined using Lowry assay.ELISA was used to detect the levels of specific IgG against rPPE65 and recombinant PstS1 protein (rPstS1) in sera from 144 patients with pulmonary tuberculosis (PTB patients), 144 health controls, and 56 patients with non-tuberculosis pulmonary diseases.ROC curves were used to determine cut-off values with the results of IgG antibodies against rPPE65 and rPstS1 for 144 PTB patients and 97 controls with negative PPD skin test.The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of rPPE65 and the combination of rPPE65 and rPstS1 were counted.Results The PPE65 protein of M.tuberculosis was successfully expressed in E.coli. The purity and concentration of rPPE65 were 95% and 0.5 mg/ml, respectively.ROC analysis showed that the cut-off of ELISA using rPPE65 was 0.64.The sensitivity, specificity, PPV, NPV, and accuracy of rPPE65 were 34.7%(50/144), 93.5%(187/200), 79.4%(50/63), 66.5%(187/287), and 68.9%(237/344), respectively.The sensitivity, specificity, PPV, NPV, and accuracy of the combination of rPPE65 and rPstS1 were 59.0%, 91.0%, 82.5%, 75.5%, 77.6%, respectively.Conclusions The rPPE65 of M.tuberculosis appears to be a candidate antigen for serodiagnosis of tuberculosis.Detection of IgG antibodies against the combination of rPPE65 and rPstS1 can increase the sensitivity of serological test for tuberculosis.

The correlation of HLA-G expression with AR and CMV active infection after kidney transplantation / 中华器官移植杂志

Objective To study the correlation of HLA-G levels with acute rejection and CMV active infection post-kidney transplantation.Methods A total of 132 initial kidney transplantation recipients were divided into kidney function stable group (F),acute rejection group (AR),CMV group according to whether they had active CMV infection and acute rejection.Forty-one healthy donors served as control group (H).HLA-G levels and mRNA expression were analyzed by using flow cytometry,ELISA,RT-PCR and Western blotting.Immunohistochemical staining was used to detect the HLA-G expression in kidney biopsies.Results The expression levels of mHLA-G1 were low in all 4 groups pre-transplantation.Only CMV group had significantly more CD14+ mHLA-G1+ cells post-transplantation (P＜0.05).sHLA-G5 levels were higher in F group than in H group (P＜0.05),but there was no significant difference among other groups pre-transplantation (P＞0.05).sHLA-G5 levels were increased significantly in CMV group as compared with F group (P＜0.05),and those in F group were higher than in H and AR groups (P＜0.05).Renal tissue biopsies from 21 renal transplantation recipients with AR indicated that HLA-G5 was expressed negatively in 17 patients,positively in 3 patients and 1 weakly positively.HLA-G was positive in the kidney tissue of 9 patients out of 9 patients with active CMV infection.In total 132 recipients,AR incidence was significantly lower in CMV ( + ) group (7.1 ％,2/28) than that in CMV ( - ) group (24.0 ％,25/104).Conclusion The sHLA-G5 may contribute to predict AR and CMV active infection; AR and CMV active infection may be correlation with immune balance in kidney transplantation recipients.

Correlation of HLA-G expression with CMV active infection following kidney transplantation / 中华器官移植杂志

ObjectiveTo determine the correlation of human leukocyte antigen-G (HLA-G)expression with CMV active infection after kidney transplantation. MethodsA total of 215 first-time kidney transplantation recipients in one transplantation center were divided into CMV ( + ) group and CMV ( - ) group according to whether they had active CMV infection. mhla-g1 expression on leukocytes was analyzed by flow cytometry. The concentrations of soluble HLA-G5 were detected by using ELISA. The sHLA-G5 cutoff levels by ROC curve was employed to predict the active CMV infection. The expression of sHLA-G5 mRNA and protein in leukocytes was analyzed by using RTPCR and Western blotting respectively. Immunohistochemical staining was used to detect the HLA-G expression in kidney biopsies of 12 cases. ResultsThe expression of mHLA-G1 in peripheral blood was low in both CMV ( + ) group and CMV ( - ) group. Also when CMV-PP65 was positive, there was no significant change in mHLA-G1. In CMV ( + ) group, the proportion of CD14+ mHLA-G1 +cells[(45. 53 ± 17.32)％]in peripheral blood was increased as compared with that in CMV (-)group[(10. 22 ± 5.78)％]. The expression of sHLA-G5 was increased significantly in CMV ( + )group. The optimal cutoff value of sHLA-G5 predicting the active CMV infection was 202. 9 μg/L,with high diagnostic accuracy. HLA-G was positive in the kidney tissue of 10 patients out of 12 patients with active CMV infection. Both RT-PCR and Western blot analysis showed that sHLA-G5 was significantly higher in CMV ( + ) group than that in CMV ( - ) group. ConclusionROC curve analysis of sHLA-G5 with the cutoff value of 202. 9 μg/L can be used to predict the active CMV infection. The HLA-G levels in peripheral blood were significantly increased and HLA-G expression in the tubular epithelial cells of the graft could be a protection mechanism of the kidney function.

Determination of cytokines in peripheral blood by cytometric bead array in kidney transplantation recipients / 中华检验医学杂志

Objective To measure the cytokines levels in peripheral blood from kidney transplantation recipients by using cytometric bead array and to analyze their change and the clinical significance in pre- and post- kidney transplantation, inducting with basiliximab and graft rejection. Methods A total of 72 renal transplantation recipients were divided into two groups, kidney function stable group(n =53) and acute rejection group (n = 19). And they were also grouped by induction with basiliximab or not,32 in basiliximab group and 40 in without basilixmab group. The levels of IFN-γ, TNF-α, IL-10, IL-5,IL-4, IL-2 were measured by cytometric bead array in peripheral blood of 72 kidney transplantation recipients and 30 healthy donors at differential time. The data was analyzed according to the following grouping:donors and recipients, kidney function stable group and acute rejection group post transplantation and with or without basiliximab group. Results The levels of TNF-α, IL-10, IL-5, IL-4, IL-2 in recipients before transplantation were ( 1.65 ±0. 10) ,(2. 55 ±0. 19) ,( 1.88 ±0. 14) ,(1.85 ±0. 12) ,(2. 12 ±0. 09) ng/L,respectively. While they were (3.04 ±0. 17), (3.33 ±0. 26), (4.03 ±0.25), (2.73 ±0. 16), (4.03 ±0. 26) ng/L respectively in healthy donors. There was statistical significance between the two groups ( t =6. 890, 2. 375, 7. 851,3.955,7.153, P<0. 01, <0. 05, <0.01, <0.01, <0.01). While the level of IFN-γ in recipients before transplantation was (2. 50 ±0. 18) ng/L,compared with (3. 00 ±0. 24) ng/L in healthy donors. There was no statistical significance between the two groups( t = 1. 625, P > 0. 05 ). The levels of IFN-γ and IL-10 in kidney function stable group were (2. 71 ± 0. 11 ) ng/L and (3.91 ± 0. 52) ng/L,while they were ( 3.30 ± 0. 36 ) ng/L and ( 12. 01 ± 5.35 ) ng/L in acute rejection group. There were statistical dirrerences between the two groups ( t = 5. 061, 11. 465, P < 0. 01, < 0. 05 ). Before induction with basiliximab, the levels of IFN-γ, TNF-α, IL-10 in recipients were (2.90 ±0. 21 ), ( 1.67 ±0. 12),(2. 45 ± 0. 16) ng/L respectively. But they were ( 2. 78 ± 0. 17 ), ( 1.58 ± 0. 07 ), ( 2. 77 ± 0. 24 ) ng/L respectively after induction with basiliximab, which showed significantly different ( t = 5. 605, 6.011,4. 126, P <0. 01, <0. 01, <0. 05). Four weeks after kidney transplantation in recipients with basiliximab,the levels of IFN-γ, IL-10, IL-4 were (2. 90 ± 0. 31 ), (9. 08 ± 0. 16), (2. 73 ± 0. 11 ) ng/L. While they were (3.28 ±0. 11 ), (4. 17 ±0. 21 ), (2. 11 ±0. 20) ng/L respectively in recipients without basiliximab induction, which were significantly different from those with basiliximab induction (t = 4. 268,4. 263,3.762, P <0. 01, <0. 01, < 0. 05 ). Conclusions Six kinds of cytokines can be measured by cytometric bead array simultaneously and accurately. The data suggests that the detection of multiple cytokines in kidney transplantation recipients by cytometric bead array can provide more guidance for clinical diagnosis and therapy.

PncA mutations in pyrazinamide-resistant Mycobacterium tuberculosis isolates / 中华检验医学杂志

Objective To evaluate the frequency of pncA gene mutations in pyrazinamide-resistant (PZA-resistant)Mycobacterium tuberculosis(M.TB)isolates.Methods The isolates were tested for PZA susceptibility with absolute concentration method.The pncA gene was amplified and the products were cloned into T-vector,followed with sequencing.Results In all the 36 M.TB isolates,there were 25 PZA-resistant strains and 11 PZA-susceptible strains.Mutations of the pncA gene were found in 10 PZA-resistant isolates,four of which belonging to high PZA-resistant strains.The wild type pncA sequence was present in 3 PZA-susceptible strains, synonymous mutations of pncA gene were detected in two PZA-susceptible strains.Additionally,11 PZA-resistant strains and 2 PZA-susceptible ones showed putative regulatory mutations in the upsteam of pncA gene.Condusions The mutation of the pncA gene can cause the PZA resistance.However.there ale other drug resistance mechanism except this mutation.

Study of rpoB Mutation in Rifampin Resistants Mycobacterium Tuberculosis / 中国医师杂志

Objective To understand the rpoB gene mutation in M.tuberculosis isolates,and to evaluate their clinical value.Method 335 clinical isolates of mycobacterium tuberculosis(109 isolates drug susceptible,246 isolates rifampin-resistance or multidrug resistance including rifampin) were detected using polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP).Results SSCP pattern of reference mycobacterium tuberculosis H37Rv as control,no mutation was found in rifampin-suscepitible 109 strains.SSCP patterns of 225/246 rifampin resistant clinical isolates were different from the normal control.The sensitivity was 91 5%.31 resistant isolates,included 25 abnormal isolates and 6 normal isolates of SCCP were identified.Sequencing showed 29 isolates had rpoB gene mutations and 3 isolates were not found rpoB gene mutations.The 3 most frequent rpoB gene mutation situs were Leu-531(19 isolates.TCGTTG) and His-526(7 isolates,CACTAC) and Asp-516(3 isolates,GACGTC).Conclusions The results confirm that rpoB gene mutation is the most important mechanism in rifampin resistant tuberculosis mutation situs are 531 tryptophan and 526 histidine;respectively.It is feasible that using PCR-SSCP to detect drug resistance in mycobacterium tuberculosis.

Cell Immunity and Physicochemical Characteristics of the Recombinant 38000 Protein of Mycobacterium tuberculosis / 中国医师杂志

Objective To evaluate the prospects of recombinant 38000 protein of Mycobacterium tuberculosis in tuberculosis epidemic investigation and subunit vaccine preparation.Methods Physicochemical characteristics of recombinant 38000 protein was detected by P I, peptide-mapping analysis and circular dichroism,guinea pig skin test,MTT stain,and peripheral blood macrophage phagocytosis were used to investigate the roles of recombinant 38000 protein in the cell immunity.Results Recombinant 38000 protein was acidic protein,its P I, was 4 67.The number of alkaline amino acid correspond with theoretic number;The secondary structure of recombinant 38000 protein was composed of ?-helix(32 6%),?-turn(31 6%) and random coil(35 8%) Recombinant 38000 protein could induce DTH in guinea pig sensitized by Mycobacterium tuberculosis Recombinant 38000 protein enhanced phagocytosis of macrophage in mice . PBMC from 30 8% healthy donors and 25% tuberculosis patients were stimulated by the recombinant 38000 protein.Conclusion Recombinant 38000 protein may be used as diagnostic reagent and as an candidate in development of subunit vaccine.

Comparison Between Extracellular Proteins of Mycobacterium Tuberculosis Virulent H37Rv and Attenuated H37Ra / 中国医师杂志

Objective To compare expression of extracellular proteins of virulent H37Rv and attenuated H37Ra in order to search differential proteins,to provide a train of thought for studing M.TB toxicity further.Methods Extracellular proteins were extracted from H37Rv and H37Ra which were inoculated and cultured on Suton's medium for three weeks.The first dimensional electrophoresis was performed on immobilized pH gradient rod gels(pH 3～10).Then the proteins in the rod gels were separated using SDS-PAGE gels.The silver-stained gels were dried and scanned with image scanner.The 2D image analysis was performed with image Master 2D Elite 3 10.Results The most protein spots deriving from extracellular proteins of H37Rv and H37Ra strains were in acidic range.In the basic range(pI more than 9 0),the number of protein spots belong to extracellular proteins of H37Rv and H37Ra was few.Three protein spots belong to low molecular range in H37Rv strain.However,absent in H37Ra strain.Conclusions Two-dimensional gel electrophoresis is useful to separate protein in Mycobacterium tuberculosis.