Objective To explore a rapid diagnostic method in neonatal sepsis and bacterial me- ningitis. Methods The primers and TaqMan probes were designed and synthesized based on the sequences of bacterial 16S rRNA gene. Nineteen bacterial strains, 3 different viruses, fungus and human genomic DNA were tested by FQ-PCR assay. Blood specimens and CSF from 195 cases of suspected septicemia were detected with both TaqMan PCR assay and blood or/and CSF culture. Results The FQ-PCR showed very high sensitivity and specificity and was able to detect at least 10 copies of 16S rRNA gene equivalent to 1~2 copies bacterium. No cross-reaction was found with human genomic DNA, other fungus and viruses. Among the 195 cases, the positive rate by FQ-PCR was 12.8%(25 cases) and 7.1%(15 cases) by blood culture ( P

Objective:To understand the laboratory characteristics for the diagnosis of immunoglobulin G4-related disease (IgG4-RD).Methods:The clinical data of 28 patients with IgG4-RD and renal damage (IgG4-RKD) diagnosed in our hospital from January 2017 to May 2019 were retrospectively analyzed. The correlation between serum IgG4 concentration and clinical features as well laboratory test results was analyzed. The 28 patients were divided into two groups: high serum IgG4 concentration group and normal serum IgG4 concentration group. The serum creatinine value, erythrocyte sedimentation rate, IgG concentration, IgA concentration, complement C3, C4 concentration, peripheral blood eosinophils, hemoglobin, IgG4/IgG and other related parameters were compared between the two groups. SPSS 20.0 statistical software was used for analysis. The two groups of measurement parameters were compared between groups by independent sample t test, non-normal measurement parameters were compared between groups by Mann-Whitney U test analysis, and the correlation between patients' IgG4 and each detection parameter was analyzed by Spearman correlation analysis. Results:Among the 28 patients, 17 were male and 11 were female, with an average age of (62±14) years. The serum IgG4 concentration increased in 75% of the patients ( n=21), with an average value of 3.01(1.41, 7.52) g/L, the serum IgG concentration increased in 64.3% of patients ( n=18), with an average value of 18.91 (12.88, 24.88) g/L, and the complement C3 decreased in 50% of the patients ( n=14), with an average value of(0.77±0.28) g/L. IgG4 was positively correlated with IgG ( r=0.422, P=0.025), IgG4/IgG ( r=0.951, P<0.01), ESR ( r=0.543, P<0.01) and peripheral blood eosinophils ( r=0.487, P<0.01), but negatively correlated with complement C3 ( r=-0.431, P=0.022) and C4 ( r=-0.504, P<0.01) levels. There were significant differences in IgG ( Z=-2.255, P=0.023), IgG4/IgG ( Z=-3.793, P<0.01), C3 ( t=7.380, P<0.01) and ESR ( t=-2.195, P=0.037) between the elevated IgG4 group and the normal group. Conclusion:Serological characteristics of IgG4-RKD combined with clinical manifestations may be able to diagnose IgG4-RKD in early stage.

Objective To explore the effects of maternal allergy symptoms on the cytokine level of umbilical cord blood of neonates.Methods A cross sectional study was conducted among 136 pregnant women in Yiwu city from 1st July to 30th December,in 2012.A questionnaire on social economic charactertistics was used and blood sam-ple of pregnant women and umbilical cord blood sample were collected to detect the level of IgE,eosinophile granulo-cyte,Eotaxin,IL-9,IL-6,IL-4,IL-5,IFN-γ,IL-10.The differences between pregnant women with and without allergy symptoms were carried out.Results There were no significant differences in social and demographic characteristics between the two groups(all P>0.05).Pregnant women with allergy symptoms had higher IgE level(0.13 IU/mL vs 0.10 IU/Ml,Z=-2.063,P=0.039),eosinophile granulocyte(0.39 ×109/mL vs 0.29 ×109/mL,Z=-2.548, P=0.011),Eotaxin(66.18ng/L vs 48.35ng/L,Z=-2.144,P=0.032),IFN-γ(927.81ng/L vs 338.65ng/L,Z=-2.051,P=0.040),IL-10(15.59ng/L vs 11.55ng/L,Z=-2.022,P=0.043) than pregnant women without allergy symptoms in neonates′cord blood.Conclusion Maternal allergy symptoms may increase the level of IgE, eosinophile granulocyte,Eotaxin,IFN-γand IL-10 of neonates′cord blood.

Objective To investigate the relationship of the clinical characteristics and the Oxford classification of IgA nephropathy.Methods Clinical presentation (age,gender,course of disease,blood pressure,hematuria,24-hour proteinuria,serum creatinine,serum albumin,triglyceride,cholesterol,and estimated glomerular filtration rate (eGFR)),pathological data (mesangial hypercellularity,endocapillary hypercellularity,segmental sclerosis or adhesions,tubular atrophy or interstitial fibrosis,artery score,and cellular + fiberocellular crescents) and their correlation of 192 patients with IgA nephropathy patients were analyzed.Results (1)Clinically,hematuria + albuminuria type was the most common among 192 the patients with IgA nephropathy (72 cases,37.5％) followed by nephrotic syndrome (42 cases,21.9％),renal insufficiency (29 cases,15.1％),hypertension (72 cases,37.5％).(2)M1 was 60.0％,E1 was 55.2％,S1 was 46.9％,T0,T1,T2 were 59.9％,22.9％,and 17.2％,respectively,small artery thickening was 46.9％,patients with cellular + fiberocellular crescents wais 48.5％.Some pathology features were related to age.(3)Proteinuria was associated with the mesangial hypercellularity score,endocapillary hypercellularity,segmental sclerosis or adhesions,and tubular atrophy or interstitial fibrosis and cellular + fiberocellular crescents.Blood pressure and renal function were associated with segmental sclerosis or adhesions,tubular atrophy or interstitial fibrosis,small artery thickening and cellular + fiberocellular crescents.Conclusions The Oxford classification has a good clinical guide of treatment and prognosis of IgA nephropathy.

Objective To explore the effects of 1α,25-dihydroxyvitamin D3 [1,25(OH)2D3] on memory CD4+T cells of focal proliferative IgA nephropathy (IgAN)patients.Methods (1) Total of twenty incipient focal proliferative IgAN patients (Lee classification:Ⅲ level) were chosen as IgAN group and 20 healthy volunteers were chosen as healthy control group.The level of serum 1,25(OH)2D3 was measured by radioimmunoassay (RIA).Peripheral blood mononuclear cells (PBMCs) were separated by the method of Ficoll density gradient centrifugation and were stimulated with anti-CD3/anti-CD28 in the absence or presence of various concentrations of 1,25(OH)2D3,Dexamethasone(DEX) and 1,25(OH)2D3and DEX combined.PBMCs were cultured for 72 hours and the levels of IFN-γ,IL-4,IL-17A,Foxp3 were measured by flow cytometry(FCM),standing for the levels of Th1,Th2,Th17,Treg.(2) IgAN group was divided into two subgroups (proteinuria ＜ 1 g/24 h subgroup,proteinuria≥ 1 g/24 h subgroup),then the serum levels of 1,25(OH)2D3,IFN-γ,IL-4,IL-17A,Foxp3 were compared.Results Compared with healthy control group,serum 1,25(OH)2D3 level of IgAN group was significantly lower (P ＜ 0.05).Serum 1,25(OH)2D3 level in proteinuria≥ 1 g/24 h subgroup was significantly lower than proteinuria ＜ 1 g/24 h subgroup and healthy control group (P ＜ 0.05).The level in proteinuria ＜ 1 g/24 h subgroup was lower than healthy control group,but the difference was not statistically significant (P ＞ 0.05).(2) The levels of IFN-γ and IL-17A and the ratios of IFN-γ/IL-4,IL-17A/Foxp3 in IgAN group increased significantly compared with healthy control group (all P ＜ 0.05),and the level of Foxp3 decreased significantly (P ＜ 0.05).The level of IL-4 also increased,but the difference was not statistically significant (P ＞ 0.05).The levels of IFN-γand IL-17A and the ratio of IL-17A/Foxp3 in proteinuria≥ 1 g/24 h subgroup increased significantly,and the level of Foxp3 decreased significantly,compared with urinary protein ＜ 1 g/24 h subgroup and healthy control group (P ＜ 0.05).The ratio of IFN-γ/IL-4 in proteinuria≥1 g/24 h subgroup and proteinuria ＜ 1 g/24 h subgroup all increased,compared with healthy control group,and the ratio in proteinuria≥ 1 g/24 h subgroup increased significantly (P ＜ 0.05).There was no significant difference in the level of IL-4 among all groups.(3) After treatment with 1,25(OH)2D3,the levels of IFN-γ and IL-17A and the ratios of IFN-γ/IL-4 and IL-17A/Foxp3 decreased significantly,and the level of Foxp3 increased significantly (P ＜ 0.05),and these effects were more obvious as the increase of the drug concentration.The level of IL-4 did not change significantly.The combination of 1,25(OH)2D3 and DEX had a synergistic inhibition on the production of IFN-γ,IL-4,IL-17A,and the ratios of IFN-γ/IL-4 and IL-17A/Foxp3,and had a synergistic promotion on the production of Foxp3.Conclusions There is a certain extent of vitamin D deficiency in focal proliferative IgAN patients,which may be associated with the severity of proteinuria.The disorder of immunomodulatory effects of memory CD4+ T cell might exist in the patients of focal proliferative IgAN.1α,25-dihydroxyvitamin D3 might have beneficial effects on the immunoregulation of memory CD4+T cells of focal proliferative IgAN patients.

Objective To investigate the effects of vitamin E (VE) on renal ischemia/reperfusion (I/R) injury of rats.Methods A total of 18 male Wistar rats were randomly divided into sham-operated group,I/R group,VE + I/R group,and each group of 6 rats.All the animals were killed at the end of 24 h of reperfusion.Nephridial tissue were examined by light microscopy,and the level of blood urea nitrogen(BUN) and serum creatinine (SCr) were measured.The protein expressions of tumor necrosis factor α (TNF-α) were detected by Western blotting.Results Compared with sham-operated group,tubulointerstitial pathological injury in I/R group was significantly aggravated,which was shown by HE and PAS stain.Compared with I/R group,the degree of morphological changes as well as renal dysfunction in VE + I/R group were obviously lessened.Meanwhile,the levels of BUN,SCr in I/R group,VE + I/R group were (10.13 ± 2.14) mmol/L and (7.67 ± 1.63) mmol/L,(80.33 ±7.15) μmol/L and (63.67 ±5.40) μ mol/L,significantly higher than those in shamed-operated group ((3.85 ± 0.21) mmol/L,(48.67 ± 3.61) μmol/L;P ＜ 0.05).And the level of BUN and SCr in VE + I/R group were significant lower than those in I/R group(P ＜0.05).Western Blotting showed that the protein expressions of TNF-α in VE + I/R group were obviously lower compared with those in group of I/R without VE treatment (P ＜ 0.05).Conclusion Vitamin E can attenuate over-expressions of TNF-α in kidney following I/R,thus protect against structural damages and renal dysfunction in I/R rat models.

Objective To investigate the mortality,hemodialysis duration and causes of death of maintenance hemodialysis patients in Shanxi province from Jan 2010 to Dec 2012.Methods The information data of Shanxi hemodialysis patients during 2010-2012 were exported from Chinese national renal data system.Mortality,hemodialysis duration and causes of death of maintenance hemodialysis patients were analyzed.Results Mortality of maintenance hemodialysis patients in Shanxi province in 2010 was 7.4％,and it was decreased to 6.4％ in 2011 and 6.8％ in 2012.Death patients with hemodialysis for less than 2 years accounted for 68.1％,and hemodialysis patients who could be maintained for more than 5 years only accounted for 8.1％.The main causes of mortality were sudden death (23.5％),cerebrovascular events (21.2％),heart failure(12.4％),myocardial infarction/ arrhythmia(6.5％),hemorrhagic disease(5.1％) and infection(4.9％).Conclusions Mortality of maintenance hemodialysis patients in Shanxi province is 6.4％ to 7.4％.Cardiovascular and cerebral vascular events are the main causes of death.

Objective To investigate the effect of adrenomedullin on rat renal tubular epithelial cell line (NRK-52E) apoptosis induced by hypoxia-reoxygenation (HR) injury and its mechanism.Methods NRK-52E cells were cultured and randomly allotted to the following 4 groups:control group,HR group,empty plasmid + HR group,ADM + HR group.NRK-52E cells were transfected with pcDNA3.1-myc-his B empty vector or pcDNA3.1-ADM by transfection complex comprising optimal proportion of plasmid and Fugene HD reagents.Cells were counted by trypanblau and cell survival rate was computed.The concentration of lactate dehydrogenase (LDH) was detected by spectrophotometric method to evaluate cell vitality.The apoptotic rate of NRK-52E cells was measured by flow cytometry.The transfer efficiency was detected by immunocytochemistry.The mRNA expressions of ADM,Bax,Bcl-2 and Fas were determined by Semi-quantitative RT-PCR,and active caspase-3,8,9 protein expression were examined by Western blotting.Results Compared with control group,the expression of ADM significantly increased in HR group (P ＜ 0.05).The expression of ADM significantly increased in ADM + HR group than that in empty plasmid + HR group.Compared with control group,in HR group,the living cell counts and cell survival rate significantly decreased; the LDH concentration in media,apoptotic rate and the levels of Bax,Bcl-2,Bax/Bcl-2,Fas,active Caspase -3,8,9 significantly increased (all P ＜ 0.05).Compared with HR group,in ADM + HR group,the living cell counts and cell survival rate significantly increased; the LDH concentration in media,the cell apoptotic rate and the levels of Bax,Bax/Bcl-2,Fas,active Caspase-3,8,9 significantly decreased,while Bcl-2 was promoted (all P ＜ 0.05).The above indexes had no differences between empty plasmid + HR group and HR group (all P ＞ 0.05).Conclusion The increased expression of ADM can inhibit NRK-52E apoptosis induced by HR,and the mechanism might be achieved by inhibiting mitochondrial and death receptor-mediated apoptotic pathways.

Objective To investigate the distribution and sequence conservation of Hap adhensin encoding gene (hap) in clinical isolates of nontypeable Haemophilus influenzae (NTHi), to screen out and identify the predominant T-and B-cell (T-B) combined antigenic epitopes on Hap protein and to analyze their immunogenicity.Methods Sequence conservation of hap genes in NTHi strains and T-B combined antigenic epitopes were predicted using bioinformatic softwares.PCR was used to amplify the 156 bp segment at 5′-end and the 855 bp segment at 3′-end of hap gene (hap-5′-156 and hap-3′-855) and the amplified products were sequenced.Phage display systems of seven T-B combined antigenic epitopes located on the 55 aa segment at N-terminal and the 285 aa segment at C-terminal of Hap protein (Hap-N52 and Hap-C285) were constructed.Western blot assay and ELISA were performed to detect the antigenicity and immunoreactivity of different T-B combined epitopes displayed by recombinant phage PⅢ protein (rPⅢ).Results Hap protein encoded by the hap gene in NTHi was located on membrane surface.Sequences of the 156 bp segment at 5′-end and the 855 bp segment at 3′-end of hap genes extracted from different NTHi strains were relatively conservative, but many mutations were found in sequences at the middle regions of these hap genes.All of the 56 NTHi strains carried hap-5′-156 and hap-3′-855 segments and shared 92.3％-100％ identities in nucleotide and amino acid sequences of these segements.Hap-N5-24 in the Hap-N52 segment as well as Hap-C4-27, Hap-C28-47, Hap-C114-129, Hap-C150-173, Hap-C200-227 and Hap-C241-267 in the Hap-C285 segment was predicted as the T-B combined antigenic epitope with a higher score and less mutations.Results of Western blot assay and ELISA confirmed that the rPⅢ-displayed Hap-C4-27 and Hap-C150-173 epitopes presented clear hybridization bands with NTHi antisera, and 96.9％ (63/65) and 92.3％ (60/65) of serum samples from children with NTHi infection were positive for antibodies against Hap-C4-27 and Hap-C150-173 epitopes, respectively.Conclusion The gene of hap is widely distributed in clinical isolates of NTHi.Moreover, sequences of the 156 pb segment at 5′-end and the 855 bp segment at 3′-end of hap gene are conservative.Hap-C4-27 and Hap-C150-173 are the predominant T-B combined antigenic epitopes on Hap protein, suggesting that they can be used as epitope candidates for developing multiple antigenic peptide vaccines against NTHi.

Aim To evaluate the effect of intermedin (IMD)on cell proliferation and regeneration in rat tu-bular epithelial cell line (NRK-52E)that was subjec-ted to hypoxia-reoxygenation (H/R)injury.Methods The NRK-52E cells were divided into control group and three model groups (H/R,H/R +primitive vec-tor,H/R +IMD vector).The content of LDH was de-tected to observe the influence of IMD on H/R injury. The cell proliferation was detected by MTT.The cell cycle was detected by flow cytometry.Real-time PCR and western blotting were used to determine mRNA and protein levels.Results ① In comparison to the con-trol,H/R treatment decreased the cell viability and in-creased LDH activity (P <0.01 );in contrast,com-pared to H/R,IMD treatment ameliorated cell viability (79.1 5 ±1 .421 % vs 61 .22 ±1 .63%,P <0.05)and decreased LDH activities by 33.85% (P <0.01 ).②The proliferation of NRK-52E cells was significantly in-hibited by H/R treatment.In comparison to the con-trol,H/R treatment of NRK-52E cells increased the proportion of cells in the G0 /G1 phase but decreased the proportion of cells in the S and G2 /M phases. Moreover,the over-expression of IMD resulted in S and G2 /Mphase redistribution and the accumulation of G2 /M-phase cells.The real-time PCR and western blotting results indicated that the mRNA and protein expression levels of cyclin D1 ,CDK4 and p57 were increased in H/R-treated cells.IMD further stimulated this up-reg-ulated expression of cyclin D1 ,CDK4 and decreased the expression of p57 in NRK-52E cells.④Cyclin D1 had a predominantly nuclear localization in NRK-52Ecells,although cytoplasmic localization was also ob-served.Conclusion The study shows that the over-expression of IMD may promote renal cell proliferation and regeneration after renal tubular cell H/R injury via the up-regulation of cyclin D1 ,CDK and the down-reg-ulation of p57.