Cancer mortality and years of life lost in Zhuanqiao Town， Minhang District of Shanghai during 2013—2017 / 上海预防医学

Objective:Death causes and life reduction of malignant tumors in the residents of Zhuanqiao Town in Minhang District from 2013 to 2017 were analyzed to provide scientific evidence for the strategies on comprehensive prevention and control of cancer. Methods:The data of death causes of malignant tumors in the residents of Zhuanqiao Town were collected and analyzed. The mortality rate， annual percent change （APC）， composition ratio， potential years of life lost （PYLL）， potential years of life lost rate （PYLLR） and average years of life lost （AYLL） of the registered population were analyzed. Results:The standardized mortality rate of malignant tumors in the residents of Zhuanqiao Town from 2013 to 2017 was 128.05/105， and the rate was higher in males than that in females. The top four cancers regarding PYLL were lung cancer， liver cancer， stomach cancer and colorectal cancer， which were roughly the same order as the top four regarding the mortality. This indicates that these four cancers had a greater impact on residents. Lung cancer had a greater impact on female life expectancy. PYLL and SPYLL ranked the first in liver cancer in males and thus had a greater impact on the males. Breast cancer was one of the most important malignant tumors in causing early death of women. Conclusion:Malignant tumor has become an important public health problem endangering the health of residents. The focus of future work in the town remains to improve public awareness of carcinogenic risk factors， actively carry out health education， lifestyle intervention and early screening in order to reduce cancer risk， alleviate cancer burden and improve the life expectancy of residents.

Acoustic radiation force impulse for evaluation of hepatic fibrosis and early stage cirrhosis / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the value of acoustic radiation force impulse (ARFI) in evaluating the stage of hepatic fibrosis and early stage cirrhosis.</p><p><b>METHODS</b>Sixty-six patients with viral hepatitis underwent liver biopsy and 33 normal subjects (S0) were selected to accept ARFI,the shear wave velocity of hepatic segments s5, s6, s7, s8 and size of liver were measured. The results of liver and spleen size and portal vein's diameter were also measured.</p><p><b>RESULTS</b>The 66 patients were divided into 3 groups: S1, S2-S3, S4. ARFI for 66 patients and 33 normal subjects showed good image quality. There were statistically significant differences between S4 group and S0 group, S1 group, S2-S3 group for the shear wave velocity of hepatic segments s5, s6, s7, s8 (P < 0.05). Between S2-S3 group and SO group S1 group, the shear wave velocity of hepatic segments s5, s6, s7, s8 also have statistically significant differences (P < 0.05), other parameters showed no significant difference (P > 0.05). Spleen size and the portal vein's diameter of S4 group were larger than those in other groups (P < 0.05).</p><p><b>CONCLUSIONS</b>The invasive acoustic radiation force impulse could evaluate the stage of hepatic fibrosis and early stage cirrhosis in patients suffering from viral hepatitis. The measurement was feasible. It was suitable for clinical use.</p>

Preliminary study of the Th17/Treg immunoregulation in patients coinfected with TB and HIV before and after HAART / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the Th17/Treg (regulatory T cells) immunoregulation in patients coinfected with TB and HIV before and after HAART(highly active anti-retroviral therapy).</p><p><b>METHODS</b>10 HIV cases coinfected with TB (HIV/TB group) and 10 cases infected with HIV only (HIV group) received HAART. PBMCs were stained and immunophenotyping of Th17 (IL-17 expressing T cells) and CD4+ CD25 T cells (Treg) were analysed by flow cytometry.</p><p><b>RESULTS</b>The pre-treatment patients tended to have lower Th17 cells and higher Tregs cells compared to post-treatment (1.90% +/- 0.9% vs. 4.65% +/- 1.48%, 16.48% +/- 4.91% vs. 8.29% +/- 3.13% respectively). The percentage of IL-17 before and after HAART were 1.90 +/- 0.9% vs. 4.65 +/- 1.48% respectively in HIV/TB group patients (P < 0.01). The difference between the percentage of IL-17 before and after HAART in the HIV/TB group and the HIV group were 2. 65 +/- 1.62% vs. 0.67% +/- 0.46% respectively (P < 0.01). IL-17 expressing T cells were increased faster after HAART in the former group than the latter. The percentage of Treg before and after HAART were 16.48% +/- 4.91% vs. 8.29% +/- 3.13% respectively in HIV/TB group (P < 0.01). The difference between the percentage of Treg before and after HAART in the HIV/TB group and the HIV group were 8.91% +/- 4.82% vs. 2.63% +/- 2.34% respectively (P < 0.01). Treg were decreased more rapidly after HAART in the former than the latter.</p><p><b>CONCLUSIONS</b>TB and HAART both had an effect on the Th17/Treg ratio of HIV/ TB co-infected patients, which can cause increased Th17 expression, the later plays a pro-inflammatory role. TB and HAART can decrease Treg expression and enhance anti-inflammation response. The fact that Th17/ Treg disorder are more likely to exist in patients with HIV/TB co-infection after HAART for one month suggests a potential role for Th17/Treg imbalance leading to tuberculosis-associated immune reconstitution inflammatory syndrome during patients receiving HAART period.</p>

The genetical evolution of the full length genes of 5 EV 71 strains from 5 Shenzhen patients with hand-food-mouth disease associated with EV71 infection / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the characteristics of molecular epidemiology and molecular evolution of 5 EV 71 (enterovirus 71, EV71) strains from 5 Shenzhen patients with hand-food-mouth disease associated with EV 71 infection.</p><p><b>METHODS</b>5 EV 71 strains were isolated, and sequenced to analyzed the full length gene sequences in order to compare nucleotide and amino acid homology with other EV71 strains from other regions and countries as well as previous strains across the world through bioinformatics software.</p><p><b>RESULTS</b>5 strains of EV 71 belonged to sub-genotype C4 by analysis of nucleotide sequences of VP1 and VP4 of EV 71. The differences of nucleotide and amino acid sequences were much small with nucleotide homology of 93% and amino acid homology of 98% among these 5 strains. A phylogenetic tree analysis indicated that 2008 Shenzhen epidemic strains were the most close to 2004 Shenzhen circulating strains, and also much close to 1998 Shenzhen epidemic strains and 2008 Fuyang Anhui strains. The dead strain was very close to 2008 Fuyang Anhui epidemic strains.</p><p><b>CONCLUSION</b>It can be speculated that this epidemic strains of EV 71 probably originate from the same ancient strain in the history, may from 1998 Shenzhen strain.</p>

Study on characteristics of cellular-mediated immune responses of novel H1N1 influenza A patients with pneumonia / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the phenotype, frequency and function of CD4+ T cell subsets and the relevant cytokines, as well as the relationship between these cells and appearance of pneumonia of novel (H1N1) influenza A patients.</p><p><b>METHODS</b>68 healthy people, 53 confirmed novel A(H1N1) influenza patients without pneumonia and 16 confirmed severe novel A (H1N1) influenza patients with pneumonia were enrolled in this study. Viral load in nasopharyngeal swabs specimens was measured by real time PCR assay. The phenotype and percentage of CD4+ T cell subsets including Th1, Th2, Th17, and Treg cells were measured by Flow cytometry analysis. The relevant cytokines in plasma including TGF-beta, IL-6 and IFN-gamma were measured by ELISA. Data was analyzed by one way ANOVA.</p><p><b>RESULTS</b>It was found that peak viral load and viral shedding period of severe patients with pneumonia was significantly increased compared with mild patients without pneumonia (P < 0.05). The percentage of Th17 cells of severe patients with pneumonia was significantly diminished compared to that of healthy subjects and mild patients without pneumonia (P < 0.05). However, Th1, Th2, Treg cells frequencies had no significant differences (P > 0.05) among these three groups. The level of TGF-beta in plasma for the severe patients with pneumonia was also significantly decreased compared to that of healthy subject and mild patients without pneumonia (P < 0.05). The viral shedding period inversely correlated with the frequency of Th17 cells (r = - 0.38, P < 0.05).</p><p><b>CONCLUSION</b>H1N1 influenza A virus can inhibit Th17 cells to differentiate, particularly more extent in patients with pneumonia. Impaired Th17 cells may correlate with viral clearance and pneumonia of novel H1N1 influenza A patients.</p>

Expression of a single-chain trimer of MHC restricted HBsAg CTL epitope using adenovirus vector containing GFP-report gene / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To generate a recombinant Adenovirus encoding a GFP (green fluorescent protein)-report gene and a single-chain trimer of MHC restricted HBsAg CTL epitope.</p><p><b>METHODS</b>An oligonucleotide encoding H-2L(d) restricted HBsAg CTL epitope was synthesized and fused with H-2L(d) DNA molecule to construct the eukaryotic expression vector carrying the HBsAg-SCT gene. The HBsAg-SCT gene was subcloned into a GFP adenovirus expression vector,which was transfected into Ad293 cells for packaging and amplification of recombinant adenovirus encoding HBsAg-SCT.</p><p><b>RESULTS</b>HBsAg-SCT has been cloned into an adenovirus vector encoding GFP report gene successfully as confirmed by double enzyme digestion and direct sequencing. HBsAg-SCT was expressed by infected Ad293 cells demonstrated by western blot assay.</p><p><b>CONCLUSION</b>A recombinant adenovirus expressing HBsAg-SCT and green fluorescent protein report gene has been generated.</p>

Association between CD4+CD25+Foxp3+ regulatory T cells and serum transforming growth factor beta 1 in patients with chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate whether the CD4+CD25+Foxp3+ regulatory T cells are associated with serum TGF beta 1 in patients with hepatitis B.</p><p><b>METHODS</b>Patients with chronic hepatitis B (CHB), chronic asymptomatic carriers (AsC), normal subjects (NS) and the resolved from HBV infection (Resolved) were recruited in this study. Flow cytometric analysis was used to detect the frequency and phenotype of peripheral CD4+CD25+Foxp3+ T cells, and Foxp3 gene expression were examined by real time PCR. Serum TGF beta 1 levels were measured by ELISA (enzyme-linked immunosorbent assay).</p><p><b>RESULTS</b>Patients with CHB or AsC exhibited significantly higher frequency of CD4+CD25+Foxp3+ T cells compared to healthy controls. CD4+CD25+ T cells derived from patients with CHB and AsC expressed higher level of Foxp3-mRNA. Furthermore, the frequency of CD4+CD25+Foxp3+ regulatory T cells was correlated with serum HBV DNA copy numbers in patients with CHB and AsC. Our results indicated that the serum TGF beta was increased in CHB and AsC patients compared to control patients, and that serum TGF beta was correlated with the expression of Foxp3-mRNA and the frequency of CD4+CD25+Foxp3+ regulatory T cells in patients with CHB and AsC.</p><p><b>CONCLUSIONS</b>The findings have important implication in the understanding of the role and mechanism of aberrant CD4+CD25+Foxp3+ regulatory T cells in the maintenance of chronicity in hepatitis B patients.</p>

Detection of cryptosporidium infection among AIDS patients in Guangdong and Yunnan / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the infection of Cryptosporidium and its epidemiological characteristics in AIDS patients of Southern China.</p><p><b>METHODS</b>Stool samples colleted from AIDS confirmed patients. The samples were detected for oocyst of Cryptosporidium by acid fast bacteria stain and indirect fluorescent antibody stain respectively, CD4 count was detected by Flow Cytometry.</p><p><b>RESULTS</b>212 samples of fresh stool obtained from the AIDS patients who live in Guangdong and Yunnan province. The total infection rate of Cryptosporidium in AIDS patients was 4.25% (9/212), the infectious rate of oocyst in the group of 50- 59-years-old was significantly higher than those in 30-39 (P < 0.01); the infectious rate of oocyst in patients with antiretroviral therapy (ART) was also significantly lower (P = 0.0000); we found the patients coinfected with Cryptosporidium with CD4 count all below 100 cells/microl. However, there were no any difference between the infectious rate to the patient's gender, areas and stool shape.</p><p><b>CONCLUSION</b>AIDS patients infected by Cryptosporidium are not rare in southern China, and the infectious rate was lower than western country. Patients received ART could decrease the infectious rate of Cryptosporidium, Cryptosporidium always happen in patient whose CD4 count was very low (< 100 cells/microl).</p>

Expression of recombinant VP1 protein of enterovirus 71 and development of serological assay for detection of EV71 infection / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To obtain a recombinant purified Enterovirus 71 VPI protein and establishment of an early, rapid and accurate serological ELISA (enzyme-linked immunosorbent assay) for detection of EV71 infection.</p><p><b>METHODS</b>VP1 gene was amplified by PCR and clonel into pET-21b (+) vector, the positive recombinant plasmid were transformed into E. coli BI21(DE3), and was induced with IPTG, the recombinant protein by SDS-PAGE and Western Blot assays. Finally, the recombinant purified VP1 protein was used as a coated antigen for detection of serum anti-IgM and IgG against EV71 by ELISA.</p><p><b>RESULTS</b>The purified VP1 was obtained, and it can be recognized by sera of patients with EV71 infection associated with hand-foot-mouth disease. The A values of anti-EV71 IgM and IgG were significantly elevated as compared to healthy objects and HFMD patients without EV71 infection (P < 0.05). The sensitivity and specificity of IgM to EV71 were 73% and 77% compared with the RT-PCR results, respectively;and those of IgG being 82% and 83%, respectively.</p><p><b>CONCLUSIONS</b>The recombinant protein VP1 was produced and purified, and it was proved to have a good antigenicity and could be used to develop a serological diagnosis kit for EV71 infection in the future.</p>

Study of the clinical and laboratory features of hand-foot-mouth disease / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the clinical and laboratory features of the mild and severe hand-foot-mouth diseases (HFMD) in Shenzhen in 2008.</p><p><b>METHODS</b>145 cases were observed in East-Lake Hospital and Shenzhen Children's Hospital. Of the 145 cases, 124 mild cases and 21 severe cases were involved.All the clinical data and laboratory findings were collected and summarized. After collection of the acute and convalescent consecutive stools and peripheral bloods from the patients with HFMDI, EV71 genes were amplified from these samples by RT-PCR. Enterovirus 71 were cultured and isolated using Vero cell line and R&D cell line.</p><p><b>RESULTS</b>The WBC counts and blood glucose levels of the severe cases were significantly elevated, but the ages of the severe ones significantly decreased compared with those of the mild cases (P < 0.05). EV71 genes could be detected by RT-PCR with 35% positive rate in mild cases and 67% in severe cases. The EV71 gene detection rate of the severe cases was significantly increased in contrast to that of the mild ones. The EV71 were isolated and cultured from the stools of 9 patients, one specimens from the dead's stool. Two severe cases died of neurogenic pulmonary edema and brain-stem encephalitis.</p><p><b>CONCLUSIONS</b>EV71 mainly contributes to HFMD and is responsible for death of some severe cases. High fever, less rash, elevated white blood cell counts and blood glucose concentrations as well as age less than 4 years old should be used for prediction of severe cases.</p>

Clinical and chest X-ray characteristics of 5 cases with severe acute respiratory syndrome in children in Shenzhen area / 中华儿科杂志

<p><b>OBJECTIVE</b>To explore clinical and chest X-ray features of SARS in children to facilitate correct diagnosis.</p><p><b>METHODS</b>Clinical manifestations and chest X-ray findings in five children suffering from SARS admitted for treatment in the hospital between February and May, 2003 in Shenzhen area were analyzed. The diagnosis was confirmed by epidemiological, clinical, laboratory and radiological examinations. Among the 5 cases, 1 was a boy and the others were girls at the age of 4 to 13 years.</p><p><b>RESULTS</b>Of the 5 SARS children, 3 presented a history of close contact with SARS patients. Fever was the initiative symptom, 4 had a body temperature of over 38 degrees C with the highest being 40 degrees C; fever sustained from 4 to 7 days with an average of 5.6 days. All the 5 cases developed nonproductive cough; on auscultation, both moist and dry rales could be heard in 3 out of the 5 cases. Mean total white count of peripheral blood was (2.96 - 6.9) x 10(9)/L, and was < 5.0 x 10(9)/L in 4 cases. SARS associated coronavirus specific RNA fragment was found positive by RT-PCR in 1 case; 1 case was positive for both IgM and IgG antibodies to the virus; 1 case was positive for only IgM antibody and another 2 cases were positive for only IgG antibody. IgG and IgM antibodies to Mycoplasma pneumoniae and Chlamydia pneumoniae as well as blood culture for bacteria were all negative. Findings on chest X-ray examination: 4 cases showed presence of patchy or macular opacities with cord-like shadows in unilateral lung plates while 1 case each showed ground-glass-like opacity and migratory changes; 1 case showed interstitial changes in the lungs in the form of irregular reticular lattice and cord-like shadows. Two cases received CT scanning and macular-patchy or spotty shadows were seen all over the lung. The shortest time for absorption of foci in the lungs was 7 days while the longest was 33 days with a mean of 15 +/- 6 days. None of the cases had any signs of fibrosis in the lungs. All the 5 cases were completely cured and discharged 7 to 40 days (mean 18 +/- 11 days) after admission.</p><p><b>CONCLUSION</b>Compared with adult cases with SARS, children with SARS had milder symptoms and signs. Presence of unilateral patchy shadow in lungs represented the main chest X-ray findings.</p>

Identification and molecular cloning and sequence analysis of a novel coronavirus from patients with SARS by RT-PCR / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the etiologic agents of the SARS and develop diagnostic method for this disease.</p><p><b>METHODS</b>Thirty-six nasopharyngeal aspirate specimens from 27 patients with SARS in Shenzhen were collected. The samples were aliquotted to three parts and subjected to molecular assays for human metapneumovirus, chlamydia and a novel coronavirus, which was reported recently to be the etiologic agent of SARS. Nested RT-PCR was used to amplify the RNA polymerase gene of the novel coronavirus and the PCR products were sequenced directly or after cloned to pMD18-T vector.</p><p><b>RESULTS</b>Human metapneumovirus and chlamydia genes were detected in none of the specimens using the RT-PCR and nested-PCR, respectively. The novel coronavirus gene were amplified in 6 of 36 specimens, the sequence analysis indicated that this novel coronavirus is unrelated to any other coronavirus reported previously. The nucleotide and deduced amino acid alignment between this coronavirus and others was not more than 40% and 70% to 82%, respectively, while the nucleotide sequence cloned from the 6 patients were identical.</p><p><b>CONCLUSIONS</b>The SARS patients in Shenzhen were infected with coronavirus and this novel coronavirus is associated with SARS. The sequence analysis indicated that the coronavirus from SARS patients in Shenzhen is the same as that identified from other areas such as Canada and Hong Kong. A specific diagnostic nested RT-PCR was developed to identify this novel coronavirus infection.</p>