Clinical study of cystatin C in assessing the severity of community acquired pneumonia / 中国基层医药

Objective To explore the clinical value of cystatin C in predicting the severity of community-acquired pneumonia(CAP).Methods From January 2016 to August 2016,the clinical data of 122 patients with CAP in Shengjing Hospital of China Medical University were retrospectively analyzed.The patients were divided into low-risk group(74 cases) and middle-high risk group (48 cases) according to pneumonia severity index (PSI).The cystatin C,procalcitonin,C-reactive protein (CRP),white blood cell (WBC) count,neutrophil percentage (NE％),D-dimer and fibrinogen(FIB) levels in hospitalized patients were detected.The relationship between cystatin C and the above related indicators and the PSI was evaluated.Draw ROC curve with cystatin C and above indicators to predict severity of CAP.Results In the low-risk group,the levels of procalcitonin,CRP,WBC count,NE％,D-dimer,FIB were[0.08 (0.04,0.23)] ng/L,[42.35(12.52,93.70)] mg/L,[7.85 (6.23,10.38)] x 109/L,(66.87 ±13.49) ％,[215.00 (134.25,410.25)] μg/L,[4.52 (3.71,5.14)] g/L,respectively,which in the middle and high risk group were [0.25 (0.07,0.54)] ng/L,[74.30 (40.45,122.75)] mg/L,[7.90 (5.78,10.63)] × 109/L,(73.97 ± 10.77) ％,[417.50 (239.75,730.00)] μg/L,[4.57 (3.87,5.08)] g/L,respectively,the differences of NE％,procalcitonin,CRP,D-dimer between the two groups were statistically significant (t =3.064,U =3.024,2.800,3.771,all P ＜0.05),the WBC count,FIB had no statistically significant differences between the two groups (all P ＞ 0.05).In the low-risk group,the level of cystatin C was [0.82 (0.68,0.91)] mmol/L,which in the middle -high risk group was [1.32 (1.12,1.54)] mmol/L,the difference between the two groups was statistically significant (U =7.978,P ＜ 0.001).When the level of cystatin C was 0.975 mmol/L,the sensitivity for the diagnosis of pneumonia severity was 95.8％,with a specificity of 85.1％.Conclusion Monitoring cystatin C levels can be used as an indicator of inflammatory response and can predict the severity of pneumonia.

Clinical study of cystatin C in assessing the severity of community acquired pneumonia / 中国基层医药

Objective@#To explore the clinical value of cystatin C in predicting the severity of community-acquired pneumonia(CAP).@*Methods@#From January 2016 to August 2016, the clinical data of 122 patients with CAP in Shengjing Hospital of China Medical University were retrospectively analyzed.The patients were divided into low-risk group(74 cases) and middle-high risk group(48 cases) according to pneumonia severity index (PSI). The cystatin C, procalcitonin, C-reactive protein(CRP), white blood cell (WBC) count, neutrophil percentage(NE%), D-dimer and fibrinogen(FIB) levels in hospitalized patients were detected.The relationship between cystatin C and the above related indicators and the PSI was evaluated.Draw ROC curve with cystatin C and above indicators to predict severity of CAP.@*Results@#In the low-risk group, the levels of procalcitonin, CRP, WBC count, NE%, D-dimer, FIB were[0.08(0.04, 0.23)]ng/L, [42.35(12.52, 93.70)]mg/L, [7.85(6.23, 10.38)]×109/L, (66.87±13.49)%, [215.00(134.25, 410.25)]μg/L, [4.52(3.71, 5.14)]g/L, respectively, which in the middle and high risk group were [0.25(0.07, 0.54)]ng/L, [74.30(40.45, 122.75)]mg/L, [7.90(5.78, 10.63)]×109/L, (73.97±10.77)%, [417.50(239.75, 730.00)]μg/L, [4.57(3.87, 5.08)]g/L, respectively, the differences of NE%, procalcitonin, CRP, D-dimer between the two groups were statistically significant(t=3.064, U=3.024, 2.800, 3.771, all P<0.05), the WBC count, FIB had no statistically significant differences between the two groups (all P>0.05). In the low-risk group, the level of cystatin C was[0.82(0.68, 0.91)]mmol/L, which in the middle-high risk group was[1.32(1.12, 1.54)]mmol/L, the difference between the two groups was statistically significant(U=7.978, P<0.001). When the level of cystatin C was 0.975 mmol/L, the sensitivity for the diagnosis of pneumonia severity was 95.8%, with a specificity of 85.1%.@*Conclusion@#Monitoring cystatin C levels can be used as an indicator of inflammatory response and can predict the severity of pneumonia.

Role of histological evaluation of periprosthetic tissue in diagnosis of periprosthetic joint infection / 中华病理学杂志

Objective@#To evaluate the role of histologicalpathology in the diagnosis of periprosthetic joint infection.@*Methods@#A total of 145 cases of joint arthroplasty during October 2017 and October 2018 from Beijing Jishuitan Hospital were collected. There were 23 cases of infection, including knee joint arthroplasty (12 cases) and hip arthroplasty (11 cases). There were 17 females and 6 males. Patients′ age ranged from 39 to 76 years (mean 63 years). The infection was diagnosed if there were >5 neutrophils per high power field in at least 5 high power field. The permanent sections were examined twice separately by two pathologists, and the interval time of histologic examination was at least two weeks. Sensitivity (SE), specificity (SP), positive predictivevalue (PPV), and negative predictive value (NPV) were calculated. The consistency evaluation of histologic examination of two pathologists was calculated by Kappa analysis.@*Results@#The neutrophil cells could locate scattered or focally in the synovium tissue of periprosthetic joint infection. Somewhere, the infiltration of vessel and the perivascular distribution could also exist. Opportunity coincidence rate between two pathologists was 91.3% (Kappa=0.817). The results showed that SE was 60.9%, SP was 100.0%, NPV was 93.1%, PPV was 100.0%.@*Conclusions@#The presence of polymorphonuclear cells in histologic examination is correlated with infection. There was high consistency between histologic examination and clinical diagnosis of joint arthroplasty.

Preparation and identification of the monoclonal antibodies against programmed death-1 ligant 1 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To prepare monoclonal antibodies (McAbs) against human programmed death-1 ligant 1(PD-L1) and identify its bioactivity.</p><p><b>METHODS</b>We immunized the BALB/c mice with Thioredoxin-(PD-L1) recombination protein which expressed by prokaryotic system. Prepare hybridoma cell by hybridoma technology and used enzyme-linked immunosorbent assay(ELISA) and Western-blotting assays to select positive hybridoma identify cell. Competition inhibition ELISA was carried out to identify the special bioactivity of antibody.</p><p><b>RESULTS</b>4 hybridoma cell strains which could secrete anti-(PD-L1) antibodies stably were selected. The McAbs has good affinity with its receptor. Purify anti-(PD-L1) with title 1:32 000 was obtained after large quantity preparation. At the same time we obtained 1 cell stain which could secret special anti-Trx McAbs.</p><p><b>CONCLUSIONS</b>We obtained anti-(PD-L1) McAbs with good bioactivity successfully, which lay the foundation for further study.</p>

Investigation on distribution of HBV genotype and serotype from Tibetan in Tongde, Qinghai / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore the distribution of HBV genotype and serotype from Tibetan in Tongde, Qinghai.</p><p><b>METHODS</b>Nested polymerase chain reaction (nPCR) was used for amplification of S gene and C gene of HBV from sera carried by Tibetan chronic HBV carrier in Tongde, Qinghai, then the HBV DNA positive products were sequenced by direct sequencing. Genotype and serotype were identified by analysis of sequence result.</p><p><b>RESULTS</b>271, which come from 311 sera samples with positive HBsAg randomly selected from natural community, were amplified and sequenced in both S gene and C gene successfully, 10 (3.7%), 261 (96.3%) out of them were identified as genotype C, recombinant between genotypes C and D respectively; 259 (95.6%), 10 (3.7%), 2 (0.7%) belonged to serotype ayw2, adr, adw2 respectively.</p><p><b>CONCLUSION</b>The recombinant between genotypes C and D was the main genotype in Tibetan chronic carrier with hepatitis Bin Tongde, Qinghai; the serotype of this areas was consisted largely of ayw2.</p>

Express the recombinant protein PD-L1 in prokaryotic and analyze its biological activity / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To construct the programmed cell death 1 ligant 1 (PD-L1) recombination expression vector, express the fusion protein in prokaryotic and analyze the biological action of express product.</p><p><b>METHODS</b>The whole PD-L1 gene sequence was synthesized after codon optimized. Construct the thioredoxin-(PD-L1) recombination expression vector and express the fusion protein in E. coli. Purified the target protein and analyze the conjugated ability of protein by ELISA.</p><p><b>RESULTS</b>The PD-L1 recombinant expression vector has been constructed correctly. The target protein has been obtained with which expressed in high efficiency and production. The target protein can conjugate specifically with the PD-1, its specific receptor.</p><p><b>CONCLUSION</b>We have obtained the PD-L1 recombinant protein success with high biological activity. The result provide the basic condition for further study on antibody and mutually action between PD-L1 and chronic virus infectious.</p>

The analysis of "alpha" dominant mutation of hepatitis B virus in community-based Zhengding area, Hebei province / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To determine the "alpha"dominant mutation of hepatitis B virus (HBV) in community-based Zhengding. Analysis the role of the newborn hepatitis B vaccination on the mutation.</p><p><b>METHODS</b>Based on the national surveillance of hepatitis B, 11,478 people's sera were collected and tested by SPRIA with kits. Collect people's sera with positive HBsAg and amplify the S gene. Sequencing and clastwaling them with the standard sequences.</p><p><b>RESULTS</b>Overall, HBV DNA was successfully amplified and sequenced in 434 of 443 samples. 6.7% samples mutated in HBV "alpha" dominant region. The difference between the mutation ratio of the two loops of HBV "alpha" dominant between the people born before and after the year 1986 has no significance.</p><p><b>CONCLUSION</b>There were HBV "alpha" dominant mutant virus in the local area with a low infection rate in the population born after the year 1986. It could not explain the newborn hepatitis B vaccination can induce the prevalence of the "alpha" dominant mutate HBV.</p>

Disequilibrium and modulation of T cell-specific transcription factors T-bet/GATA-3 in CD4+T cells from ovalbumin-sensitized mice / 中华微生物学和免疫学杂志

Objective To identify the imbalance of T cell-specific transcription factors T-bet/GATA-3,and to explore the modulation with dexamethasone and imiquimod in CD4+T cells from ovalbumin (OVA)sensitized mice.Methods CD4+T cells were obtained fromsingled-cell suspension of spleen(after lysis of RBC).ELISA assay was used to detect the concentrations of IL-4,IL-5 and IFN-γin superna tants and cell pellets,and the expression of T-bet and GATA-3 was detected by Western blot.Resuits In the control group,tIle low levels of IFN-γ were detected in the supernatants during 24 h.In OVA treatment group,the concentrations of IL-4,IL-5 were increased significantly,and the concentrations of IFN-γ were always low in the supernatants.In the dexamethasone treatment group,the concentrations of IFN-γ,IL-4 and IL-5 were all low in the supernatants during 24 h.In the imiquimod treatment group,the concentrations of IFN-γ were increased significantly,and the concentrations of IL-4 and IL-5 were decreased in the super natants.It worked at 6 h,and achieved the peak at 12 h,lasted over 24 h.In the control group,the expres sions of T-bet and GATA-3 were detected in CD4+T cells during 24 h.In OVA treatment group,the expressions of T-bet were decreased,and that of GATA-3 were increased rapidly in CD4+T cells.In dexam ethasone treatment group,the expressions of T-bet were always low in CD4+T cells,and that ofGATA-3 were no change during 24 h.In imiquimod treatment group,the expressions of T-bet were increased,andthat of GATA-3 were decreased in CD4+T cells.The protein expressions worked at 6 h.and achieved the peak at 12 h,lasted over 24 h.Conclusion The imbalance T cell-specific transcription factors T-bet/GA-TA-3 contributes to both high expression of GATA-3 and low expression of T-bet in CD4+T cells from OVA sensitized mice.Dexamethasone treatment inhibits the expression of T-bet in CD4+T cells and has no func tion in GATA-3.Imiquimod treatment modulates key master switches GATA-3 and T-bet that results in com mitting T helper cell to a TH 1 phenotype and imiquimod may play a key role in the regulation of TH2 cytokine responses in asthma.

The analysis of hepatitis B virus genetic characterization and ratio of mutation in low-age group of HuNan / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>In order to investigate the characterization of mutation and genotype distributing in the younger group which was under the universal vaccination. The sequence of HBV was analyzed to offer the information to control and prevention in the area.</p><p><b>METHODS</b>Young person's sera with positive HBsAg are collected, and the Large S sequence of HBV including preS and S gene are amplified and sequenced. The genotype and serotype were determined by clastwal with the standard genotype sequence. And one virus complete genome is amplified.</p><p><b>RESULTS</b>The virus gene are successful amplified from the 33 sera. The sequence result indicate the 30 of 33 (90.9%) HBV genotype is B and 3 of 33 (9.0%) is C. The HBV serotype including ayw (1), adr (3), adw (29), 5 of 33 mutated in the "a" dominant of HBV, and the percentage is 15.2% . The HBV full length gene of serum number of 5856 is amplified and sequenced. Its genotype is B, serotype is adw and length is 3215 base.</p><p><b>CONCLUSIONS</b>The dominant genotype of HuNan is B, and the dominant serotype is adw.</p>

Detection of HBV DNA among HBsAg negative children after prevention for mother-to-infant transmission / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Detection HBV DNA among HBsAg negative children who have been vaccined at birth, in order to improve the evaluation of the indicator for HBV DNA infection.</p><p><b>METHODS</b>Selection HBsAg negative children who have been vaccined at birth and then detection HBV DNA from sera using QIAamp Viral DNA Mini Kit, HBV DNA s region was obtained by nested PCR and sequencing.</p><p><b>RESULTS</b>12 of the 140 children were HBV DNA detected were positive and the infectious rate was 8.6% . No mutant of the 12 HBV DNA in "a" determinant.</p><p><b>CONCLUSION</b>To evaluate the effection of the prevention of HBV mother-to-child transmission, the standard method should be established. The detection of HBV DNA should be included in the future.</p>

Cloning of the full-length genome of A recombinant hepatitis B virus strain and phylogenetic analysis / 病毒学报

An aberrant genotype of hepatitis B virus was discovered from a female child when we surveyed the status of the virus' infection in Guangxi of China. The full-length genome was amplified and sequenced. The length of genome is 3215 bp and the serotype of the virus is adr. In phylogenetic tree analysis with the standard genotype sequence of GenBank, the genome was clustered with genotype C, however, phylogenetic tree analysis of the individual segment supported recombination strain was formed. The segment between nt 1630 and 2880 was similar to genotype C, and the other part of genome close to genotype A. The result suggests it is a recombinant virus strain. The finding provides a reference to study the genotype and evolution of hepatitis B virus in China.

Prokaryotic expression and purification of human papillomavirus type 11 L2E7 fusion protein vaccine and its immunnogenicity / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To construct the Escherichia coli (E. coli) prokaryotic expression system pET9aHPV11L2E7, purify the fusion protein L2E7 and study the immunnogenicity of the protein.</p><p><b>METHODS</b>The HPV11 L2, E7 coding region was amplified from condyloma acuminata tissue specimen by PCR. The recombinant plazmid pET9aHPV11L2E7 was established and sequenced. Fusion protein L2E7 (553 amino acids) was expressed in host strain BL21 (DE3plus) by IPTG inducing and identified by using SDS-PAGE and Western blotting. Then L2E7 protein purified with CM column was inoculated to Balb/c mice and its cell-mediated and humoral immunnogenicity was assessed by IFN-gamma enzyme-linked immunospot (ELISPOT) and enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The E. coli prokaryotic expression system pET9aHPV11L2E7 was established and the purified fusion protein L2E7 was obtained successfully. The mice in vivo experiment indicated that the purified protein L2E7 could induce HPV11E7 specific cell-mediated immune responses and high level HPV L2E7 antibody was detected in serum.</p><p><b>CONCLUSION</b>The purified fusion protein L2E7 could induce specific cell-mediated and humoral immune responses. It can be used as a candidate of genital wart immune therapeutic vaccine.</p>

Expression of the human papillomavirus type 16L/E7 fusion protein in E. coli and observation of its immunogenicity in mice / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>Many epidemiological and experimental evidences prove that cervical cancers are strongly associated with genital high-risk types of human papillomavirus (HPV). HPV16 is present in 50% of the tumor specimens. Thus, it is important to develop vaccines against HPV16 and cervical cancer. The authors studied the expression of the HPV16 L1DeltaCE7N fusion protein in E. coli and observed its immunogenicity.</p><p><b>METHODS</b>The fragment of HPV16 L1DeltaC gene and the E7N gene were amplified by PCR separately; the fusion gene named L1DeltaCE7N was generated by fusing E7N to the C terminal of L1DeltaC then the chimeric gene was cloned into prokaryotic expression vector pGEX-2T and expressed in E. coli strain JM109. The L1DeltaCE7N protein expressed were detected by Western blot. Finally its immunogenicity was characterized in immunized mice.</p><p><b>RESULTS</b>It was proved that the sequence and open reading frame of fusion gene L1DeltaE7N was correct by sequencing; SDA-PAGE gel analysis showed that HPV16 L1/E7 fusion protein was highly expressed in E. coli; the protein was expressed as soluble form and the molecular weight was about 85 x 10(3). The fusion protein could be purified by affinity chromatography and gel filtration. The ELISA result indicated that L1/E7 could elicit specific antibodies against L1 and E7 in immunized mice. In vivo tumor protection test indicated that tumor formation was retarded or prevented in the mice after vaccination with L1/E7, when C57 BL/6 mice were challenged by syngeneic HVP16E6 and E7 transformed tumor cells.</p><p><b>CONCLUSION</b>HPV16L1/E7 fusion protein was expressed in E. coli, it can be a candidate for prophylactic and therapeutic vaccine for HPV16-associated infection and tumors.</p>

Treatment of allergic airway inflammation and hyperresponsiveness by imiquimod modulating transcription factors T-bet and GATA-3 / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Imiquimod is an imidazoquinoline, which class of compounds are known to have antiviral and antitumoural properties. In recent studies, it was shown that imiquimod modulates the T helper cell type Th1/Th2 response by inducing the production of Th1 cytokines like IFN-gamma, and by inhibiting the Th2 cytokines like interleukin (IL)-4. Several investigators have shown that T-bet and GATA-3 are master Th1 and Th2 regulatory transcription factors. This study investigated whether imiquimod treatment inhibited airway inflammation by modulating transcription factors T-bet and GATA-3.</p><p><b>METHODS</b>Thirty-six male SD rats were randomly divided into a control group, an asthmatic group, and an imiquimod group, which was exposed to an aerosol of 0.15% imiquimod. Twenty-four hours after the last ovalbumin (OVA) challenge, airway responsiveness was measured and changes in airway histology were observed. The concentrations of IL-4, IL-5 and IFN-gamma in bronchoalveolar lavage fluid (BALF) and serum were measured by enzyme linked immunosorbent assay (ELISA). The mRNA expressions of IL-4, IL-5, IFN-gamma, T-bet and GATA-3 in lung and in CD4(+) T cells were determined by reverse transcription polymerase chain reaction (RT-PCR). The protein expressions of T-bet and GATA-3 were measured by Western blot.</p><p><b>RESULTS</b>It was demonstrated that imiquimod 1) attenuated OVA induced airway inflammation; 2) diminished the degree of airway hyperresponsiveness (AHR); 3) decreased the Th2 type cytokines and increased Th1 type cytokines mRNA and protein levels; 4) modulated the Th1/Th2 reaction by inhibiting GATA-3 production and increasing T-bet production.</p><p><b>CONCLUSION</b>Imiquimod treatment inhibits OVA induced airway inflammation by modulating key master switches GATA-3 and T-bet that result in committing T helper cells to a Th1 phenotype.</p>

Construction and identification of non-replication recombinant vaccinia virus co-expressing human papillomavirus type 16 L1/L2/E6/E7 proteins / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To generate a human papillomavirus (HPV16) prophylactic and therapeutic vaccine candidate for cervical cancer.</p><p><b>METHODS</b>HPV16 major capsid protein L1 gene/minor capsid protein L2 gene and HPV16 early E6/E7 genes were inserted into a vaccinia virus expression vector. A strain of non-recombinant vaccinia virus containing the sequences was obtained through a homologous recombination and identified.</p><p><b>RESULTS</b>DNA hybridization confirmed that the HPV16L1/L2/E6/E7 genes were integrated into vaccinia virus DNA. Western Blot result showed that full-length L1/L2/E6/E7 proteins were co-expressed in CEF cells infected with the recombinant virus.</p><p><b>CONCLUSION</b>NTVJE6E7CKL1L2 could be taken as a candidate of prophylactic and therapeutic vaccine for HPV-associated tumors and their precancerous transformations.</p>