Metformin Mitigates Sepsis-Induced Acute Lung Injury and Inflammation in Young Mice by Suppressing the S100A8/A9-NLRP3-IL-1ß Signaling Pathway.

Background: Globally, the subsequent complications that accompany sepsis result in remarkable morbidity and mortality rates. The lung is among the vulnerable organs that incur the sepsis-linked inflammatory storm and frequently culminates into ARDS/ALI. The metformin-prescribed anti-diabetic drug has been revealed with anti-inflammatory effects in sepsis, but the underlying mechanisms remain unclear. This study aimed to ascertain metformin's effects and functions in a young mouse model of sepsis-induced ALI. Methods: Mice were randomly divided into 4 groups: sham, sham+ Met, CLP, and CLP+ Met. CLP was established as the sepsis-induced ALI model accompanied by intraperitoneal metformin treatment. At day 7, the survival state of mice was noted, including survival rate, weight, and M-CASS. Lung histological pathology and injury scores were determined by hematoxylin-eosin staining. The pulmonary coefficient was used to evaluate pulmonary edema. Furthermore, IL-1ß, CCL3, CXCL11, S100A8, S100A9 and NLRP3 expression in tissues collected from lungs were determined by qPCR, IL-1ß, IL-18, TNF-α by ELISA, caspase-1, ASC, NLRP3, P65, p-P65, GSDMD-F, GSDMD-N, IL-1ß and S100A8/A9 by Western blot. Results: The data affirmed that metformin enhanced the survival rate, lessened lung tissue injury, and diminished the expression of inflammatory factors in young mice with sepsis induced by CLP. In contrast to sham mice, the CLP mice were affirmed to manifest ALI-linked pathologies following CLP-induced sepsis. The expressions of pro-inflammatory factors, for instance, IL-1ß, IL-18, TNF-α, CXCL11, S100A8, and S100A9 are markedly enhanced by CLP, while metformin abolished this adverse effect. Western blot analyses indicated that metformin inhibited the sepsis-induced activation of GSDMD and the upregulation of S100A8/A9, NLRP3, and ASC. Conclusion: Metformin could improve the survival rate, lessen lung tissue injury, and minimize the expression of inflammatory factors in young mice with sepsis induced by CLP. Metformin reduced sepsis-induced ALI via inhibiting the NF-κB signaling pathway and inhibiting pyroptosis by the S100A8/A9-NLRP3-IL-1ß pathway.

Radiomic signatures reveal multiscale intratumor heterogeneity associated with tissue tolerance and survival in re-irradiated nasopharyngeal carcinoma: a multicenter study.

BACKGROUND: Post-radiation nasopharyngeal necrosis (PRNN) is a severe adverse event following re-radiotherapy for patients with locally recurrent nasopharyngeal carcinoma (LRNPC) and associated with decreased survival. Biological heterogeneity in recurrent tumors contributes to the different risks of PRNN. Radiomics can be used to mine high-throughput non-invasive image features to predict clinical outcomes and capture underlying biological functions. We aimed to develop a radiogenomic signature for the pre-treatment prediction of PRNN to guide re-radiotherapy in patients with LRNPC. METHODS: This multicenter study included 761 re-irradiated patients with LRNPC at four centers in NPC endemic area and divided them into training, internal validation, and external validation cohorts. We built a machine learning (random forest) radiomic signature based on the pre-treatment multiparametric magnetic resonance images for predicting PRNN following re-radiotherapy. We comprehensively assessed the performance of the radiomic signature. Transcriptomic sequencing and gene set enrichment analyses were conducted to identify the associated biological processes. RESULTS: The radiomic signature showed discrimination of 1-year PRNN in the training, internal validation, and external validation cohorts (area under the curve (AUC) 0.713-0.756). Stratified by a cutoff score of 0.735, patients with high-risk signature had higher incidences of PRNN than patients with low-risk signature (1-year PRNN rates 42.2-62.5% vs. 16.3-18.8%, P < 0.001). The signature significantly outperformed the clinical model (P < 0.05) and was generalizable across different centers, imaging parameters, and patient subgroups. The radiomic signature had prognostic value concerning its correlation with PRNN-related deaths (hazard ratio (HR) 3.07-6.75, P < 0.001) and all causes of deaths (HR 1.53-2.30, P < 0.01). Radiogenomics analyses revealed associations between the radiomic signature and signaling pathways involved in tissue fibrosis and vascularity. CONCLUSIONS: We present a radiomic signature for the individualized risk assessment of PRNN following re-radiotherapy, which may serve as a noninvasive radio-biomarker of radiation injury-associated processes and a useful clinical tool to personalize treatment recommendations for patients with LANPC.

Development and validation of a score predicting mortality for older patients with mitral regurgitation / 老年心脏病学杂志（英文版）

OBJECTIVE@#To develop and validate a user-friendly risk score for older mitral regurgitation (MR) patients, referred to as the Elder-MR score.@*METHODS@#The China Senile Valvular Heart Disease (China-DVD) Cohort Study functioned as the development cohort, while the China Valvular Heart Disease (China-VHD) Study was employed for external validation. We included patients aged 60 years and above receiving medical treatment for moderate or severe MR (2274 patients in the development cohort and 1929 patients in the validation cohort). Candidate predictors were chosen using Cox's proportional hazards model and stepwise selection with Akaike's information criterion.@*RESULTS@#Eight predictors were identified: age ≥ 75 years, body mass index < 20 kg/m2, NYHA class III/IV, secondary MR, anemia, estimated glomerular filtration rate < 60 mL/min per 1.73 m2, albumin < 35 g/L, and left ventricular ejection fraction < 60%. The model displayed satisfactory performance in predicting one-year mortality in both the development cohort (C-statistic = 0.73, 95% CI: 0.69-0.77, Brier score = 0.06) and the validation cohort (C-statistic = 0.73, 95% CI: 0.68-0.78, Brier score = 0.06). The Elder-MR score ranges from 0 to 15 points. At a one-year follow-up, each point increase in the Elder-MR score represents a 1.27-fold risk of death (HR = 1.27, 95% CI: 1.21-1.34, P < 0.001) in the development cohort and a 1.24-fold risk of death (HR = 1.24, 95% CI: 1.17-1.30, P < 0.001) in the validation cohort. Compared to EuroSCORE II, the Elder-MR score demonstrated superior predictive accuracy for one-year mortality in the validation cohort (C-statistic = 0.71 vs. 0.70, net reclassification improvement = 0.320, P < 0.01; integrated discrimination improvement = 0.029, P < 0.01).@*CONCLUSIONS@#The Elder-MR score may serve as an effective risk stratification tool to assist clinical decision-making in older MR patients.

Differentiating pediatric cystic nephroma from common renal multicystic lesions: A case report.

Pediatric cystic nephroma is a rare, clinically benign, renal tumor. Pediatric renal cystic lesions are complex. Imaging findings and tumor appearance are often nonspecific, and careful pathological examination is necessary. We discuss diagnosis of pediatric cystic nephroma and how to differentiate it from multicystic dysplastic kidney and cystic partially differentiated nephroblastoma.

Long-term Survivals, Toxicities and the Role of Chemotherapy in Early-Stage Nasopharyngeal Carcinoma Patients Treated with Intensity-Modulated Radiation Therapy: A Retrospective Study with 15-Year Follow-up / Journal of the Korean Cancer Association, 대한암학회지

Purpose@#This study was aimed to investigate long-term survivals and toxicities of early-stage nasopharyngeal carcinoma (NPC) in endemic area, evaluating the role of chemotherapy in stage II patients. @*Materials and Methods@#Totally 187 patients with newly diagnosed NPC and restaged American Joint Committee on Cancer/ International Union Against Cancer 8th T1-2N0-1M0 were retrospectively recruited. All received intensity-modulated radiotherapy (IMRT)±chemotherapy (CT) from 2001 to 2010. @*Results@#With 15.7-year median follow-up, 10-year locoregional recurrence-free survival, distant metastasis-free survival (DMFS), disease-specific survival (DSS), and overall survival (OS) were 93.3%, 93.5%, 92.9% and 88.2%, respectively. Multivariable analyses showed cervical lymph nodes positive and pre-treatment prognostic nutritional index ≥ 52.0 could independently predict DMFS (p=0.036 and p=0.011), DSS (p=0.014 and p=0.026), and OS (p=0.002 and p 45 years (p=0.002) and pre-treatment lactate dehydrogenase ≥ 240 U/L (p 0.05). Unsurprising, patients in IMRT+CT had more acute gastrointestinal reaction, myelosuppression, mucositis, late ear toxicity, and cranial nerve injury (all p < 0.05) than IMRT alone group. @*Conclusion@#Superior tumor control and satisfying long-term outcomes could be achieved with IMRT in early-stage NPC with mild late toxicities. As CT would bring more toxicities, it should be carefully performed to stage II patients.

Survey of the record filing situations of Shenzhen biosafety laboratories in 2017 / 中华医学科研管理杂志

Objective Aims to reduce the hidden risks of laboratory biosafety,understand the status of biosafety laboratory record filing situations in Shenzhen,and also to provide scientific basis for further standardizing the management of biosafety laboratory in Shenzhen.Methods In 2017,75 laboratories in Shenzhen completed record filing were surveyed,method ologies adopted including application materials review,phone call consultation and communication,carrying out corrective ac tions based on feedback peer review suggestions and finally complete the record filing.Results The first/second level laborato ry of biosafety in shenzhen is mainly public medical institutions,followed by private hospitals.In 2017,the first three recordfiling LABS were Futian district,nanshan district and longgang district.According to the data analysis,lack of the second category of pathogenic microorganism laboratory activity project risk assessment report,and laboratory layout diagram function partition is not clear were two of the more prominent problems in the software and hardware of laboratory management respectively.Conclusions Basically,the overall record filing of Shenzhen biosafety laboratory is good,however,more measurements should be developed to deal with identified problems to further strengthen the standardized management of laboratory biosafety.

Identification and characterization of MYC transcription factors in Taxus sp.

Myelocytomatosis (MYC) transcription factors (TFs) are key regulators of the jasmonic acid (JA) signaling pathway. In cell cultures, methyl jasmonate (MeJA) can improve the production of taxol, which is a complex terpenoid compound with an intense antitumor activity. However, the functions of MYC genes in Taxus sp. (yew trees) remain poorly known. Based on Taxus sp. transcriptome changes induced by MeJA, a TcMYC gene was isolated in a previous study. Here, we further characterized the TcMYC TF encoded by that gene and four other yew MYC TFs previously obtained. Three yew MYC TFs had the typical basic helix-loop-helix (bHLH)-MYC_N region, but the other two MYC did not, although all five presented the bHLH domain. TcMYC was localized to the nuclei, and phylogenetic analysis indicated that the yew MYC TFs were closely related to Arabidopsis thaliana MYC1/2 and maize R protein. The yeast one-hybrid assay showed that TcMYC binds the G-box of the promoter of taxane 5α-hydroxylase. Transcript levels of TcMYC revealed that TcMYC was highly expressed in xylem and leaves, and up-regulated by drought and high-salinity stresses. Coronatine (COR) has recently been used as a new elicitor to improve the production of taxol in cell cultures; TcMYC was strongly expressed at 2 and 4 h after COR treatment, but decreased at 12 and 24 h. Overall, the results obtained here provide new insights into the potential regulatory roles of MYC TFs on taxol biosynthesis in yew trees.

Differential proteomic analysis for immune complexes deposited in synovial tissues of RA patients using liquid chromatography-tandem mass spectrometry / 临床检验杂志

Objective To scan protein expression profile of immune complexes (ICs) derived from the synovial tissue of the patients with rheumatoid arthritis (RA) based on liquid chromatography-tandem mass spectrometry (LC-MS).Methods The samples of synovial fluid were obtained from knee joints of the patients with RA and osteoarthritis (OA) used as control during therapeutic arthrocentesis in knee jiont at the Department of Orthopedics of Jinling Hospital,School of Medicine,Nanjing University.The protein expression profile of ICs was identified by enrichment strategy based on immunoprecipitation and LC-MS analysis.The value of fraction of total (FOT) was used to estimate protein abundance and screen the up-and down-regulated proteins.The function enrichment,interaction network and signal pathway of differential proteins were analyzed using softwares David and String.Results A total of 511 and 526 protein spots in ICs of RA and OA patients were identified respectively.Among them,170 proteins existed only in RA group.45 and 85 proteins in RA group were statistically up-and down-expressed compared with controls.Conclusion HSP90AA1,HSP70,HLAG,Thioredoxin,Annexin A2 and vitronectin may be involved in the pathogenesis of RA through different paths and possible to become promising diagnostic indicators or new therapeutic targets for RA.

Food-advanced glycation end products aggravate the diabetic vascular complications via modulating the AGEs/RAGE pathway / 中国天然药物

The aim of this study was to investigate the effects of high-advanced glycation end products (AGEs) diet on diabetic vascular complications. The Streptozocin (STZ)-induced diabetic mice were fed with high-AGEs diet. Diabetic characteristics, indicators of renal and cardiovascular functions, and pathohistology of pancreas, heart and renal were evaluated. AGEs/RAGE/ROS pathway parameters were determined. During the experiments, the diabetic mice exhibited typical characteristics including weight loss, polydipsia, polyphagia, polyuria, high-blood glucose, and low-serum insulin levels. However, high-AGEs diet effectively aggravated these diabetic characteristics. It also increased the 24-h urine protein levels, serum levels of urea nitrogen, creatinine, c-reactive protein (CRP), low density lipoprotein (LDL), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) in the diabetic mice. High-AGEs diet deteriorated the histology of pancreas, heart, and kidneys, and caused structural alterations of endothelial cells, mesangial cells and podocytes in renal cortex. Eventually, high-AGEs diet contributed to the high-AGE levels in serum and kidneys, high-levels of reactive oxygen species (ROS) and low-levels of superoxide dismutase (SOD) in serum, heart, and kidneys. It also upregulated RAGE mRNA and protein expression in heart and kidneys. Our results showed that high-AGEs diet deteriorated vascular complications in the diabetic mice. The activation of AGEs/RAGE/ROS pathway may be involved in the pathogenesis of vascular complications in diabetes.

Principal Items Respond to Activities of Daily Living for Persons with Physical Disability / 中国康复理论与实践

@#Objective To investigate the principal indicators of self-care ability in activities of daily living for the persons with physical disability. Methods Persons with physical disability were asked to select the top 3 items out of 27 items of activities of daily living in 3 levels. Results A total of 1960 questionnaires were send out, and 1862 were returned. For all the subjects, the items related with personal hygiene, such as toileting, self-cleaning and bathing, were selected 899 times (16.1%). The items related with personal health, as visiting community clinics and community exercising, were selected 570 times (10.2%). The items related with social interaction, as making a telephone and chatting, were selected 500 times (9.0%). For the persons with physical disability of first grade, the major items most selected were eating, entertaining, self-cleaning and transferring; and self-cleaning and housework for those of second grade; self-cleaning, community activities and housework for those of third grade; and community and interaction for those of fourth grade. Conclusion The persons with physical disability mostly focused on the activities related with personal hygiene, health and social interaction, and varied with the severity of disability, from self-cleaning to housework and social participation.

Chemokine Signaling Pathway Involved in CCL2 Expression in Patients with Rheumatoid Arthritis

PURPOSE: Rheumatoid arthritis (RA) is an inflammatory joint disorder, the progression of which leads to the destruction of cartilage and bone. Chemokines are involved in RA pathogenesis. In this study, we investigated the chemokine signaling pathway associated with CCL2 in peripheral blood (PB) and synovial tissues (ST) of RA patients based on our previous work about chemokine signaling pathway involved in the activation of CCL2 production in collagen-induced arthritis rat ST. MATERIALS AND METHODS: Total RNA was isolated from PB leukocytes and synovium of the knee joint in both RA patients and control populations. Real-time polymerase chain reaction was used to determine CCL4, CCR5, c-Jun, c-Fos, and CCL2 expressions. Serum level of CCL2 was assessed by enzyme-linked immunosorbent assay, and the production of CCL2 in ST was analyzed immunohistochemically. RESULTS: The expressions of CCL4, CCR5, c-Jun, c-Fos, and CCL2 messenger RNA in RA patients were significantly higher than those in healthy controls, both in ST and on PB leukocyte. Serum CCL2 levels were elevated in RA patients. Histological examination of rheumatoid joints revealed extensive CCL2 expression in RA ST. CONCLUSION: CCL2, CCL4, c-Jun, c-Fos, and CCR5 may play an important role in the recruitment of PB leukocytes into the RA joints. These data provide evidence that the chemokine signaling pathway is involved in CCL2 expression in RA patient tissues, which may contribute to chronic inflammation associated with RA. Targeting this signaling pathway may provide a novel therapeutic avenue in RA.

Protection of morroniside on STZ-induded diabetic nephropathy aggravated by AGEs in mice and its mechanism / 中草药

Objective: To observe the protective mechanism of morroniside (an active component in Cornus officinalis) on the diabetic nephropathy (DN) mice induced by streptozocin (STZ) and aggravated by advanced glycation end products (AGEs). Methods: The DN mice were fed with high-AGEs fodders, and ig administered with aminoguanidine (0.1 g/kg), metformin (0.2 g/kg), captopril (0.02 g/kg), low-dose morroniside (0.02 g/kg), and high-dose morroniside (0.10 g/kg) for 12 weeks. After that, the fasting glucose, insulin, serum creatinine, urea nitrogen, serum and renal AGEs, 24 h urine protein, etc were measured, the RT-PCR method was used to detect the levels of receptor for advanced glycation end products (RAGE) mRNA, the Western blotting technique was used to test the protein expression levels, and the pathologic changes of mice pancreas and kidney were observed. Results: Morroniside could significantly decrease the fasting blood glucose levels, alleviate the symptoms of polydipsia, polyphagia, polyuria, and weight loss, increase the insulin production, and reduce the levels of 24 h urine protein, serum urea nitrogen, creatinine, serum and renal AGEs. Furthermore, morroniside could also anesis the lesions of pancreas and kidney and reduce the levels of RAGE mRNA and protein expression in renal cortex. Conclusion: Morroniside has the protactive effects on DN mice and high-dose morroniside was much better. The mechanism may be related to the reduced levels of AGEs and RAGE mRNA and protein expression.

Left ventricular function in newborn infants of mothers with gestational diabetes mellitus / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To evaluate left ventricular function in newborn infants of mothers with gestational diabetes mellitus (GDM).</p><p><b>METHODS</b>Forty newborn infants of mother with GDM (GDM group) and forty normal newborn infants (control group) were enrolled in this study. Two-dimensional speckle tracking imaging was used to measure interventricular septal thickness, posterior left ventricular wall thickness and left ventricular ejection fraction in both groups. Left ventricular rotation and torsion were evaluated for all participants.</p><p><b>RESULTS</b>Interventricular septal thickness in the GDM group was much higher than in the control group (0.45±0.06 mm vs 0.34±0.05 mm; P<0.05). Posterior left ventricular wall thickness in the GDM group was also higher than in the control group (0.45±0.17 mm vs 0.31±0.02 mm; P<0.05). There was no difference in the left ventricular ejection fraction between the two groups (P>0.05). Peak subendocardial rotation, peak subepicardial rotation, peak bulk rotation and peak mural torsion were higher in the GDM group than in the control group (P<0.05).</p><p><b>CONCLUSIONS</b>Cardiac function may be impaired in newborn infants of mothers with GDM, with changes in left ventricular shape and abnormalities of left ventricular rotation and torsion. However, infants have a normal ventricular blood ejection under the cardiac compensation. Two-dimensional speckle tracking imaging technique can be used for early detection of left ventricular function.</p>

Clinical analysis of early surgical operation in infective endocarditis / 中华胸心血管外科杂志

Objective The surgery time for patients with infective endocarditis (IE) has been transformed.It has been supported that,for the patients with surgical indications,the surgery time should be as early as possible to improve the clinical outcome.The purpose of the research is to identify whether the patients with IE could get further benefit from early surgery.Methods Between June 1996 and July 2011,135 IE patients'data has been collected retrospectively,all of whom were verified through the modified Duke categories.The patients were devided into group A( the new therapeutic schedule group after 2008 ) and group B( the traditional therapeutic schedule group before 2008 ) by the year of 2008.The end points of observation were death associated with IE,cardiac failure,embolism,and re-infection.The comparison between the groups was by means of non-parameter rank and inspection test,variance analysis,t test,chi-square test,fisher exact test.The outcome comparison between the groups was via the Kaplan-Meier survival analysis.Results There were no significant differences in baseline data between the groups.No survival differences could be observed via the Kaplan-Meier analysis( Log Rank P =0.189).During the following-up visit,compared with the patients in group B,the mortality in group A is lower(9.4％ vs.23.0％,P=0.016),the incidence of heart failu re was less in group A (5.4％ vs.26.2％,P ＜0.001 ).No differences could be found in re-infection between the two groups(0 vs.4.9％,P =0.112 ). More patients in group A underwent surgery (67.6％ vs.32.8％,P ＜0.001 ).Conclusion The new therapeutic sehedule of IE coull reduce the mortality rate and promote the cardiae funetion.The incidence of re-infeetion didn't increase.

Applicability of a sensitive duplex real-time PCR assay for identifying B/Yamagata and B/Victoria lineages of influenza virus / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To develop a novel sensitive duplex real-time PCR assay for accurately identifying B/Yamagata and B/Victoria lineages of influenza virus type B.</p><p><b>METHODS</b>50 HA (hemagglutinin) gene sequences coding for B/Yamagata and B/Victoria lineage, respectively, were randomly downloaded for GenBank and analyzed by software MEGA. Primers and probes specific for HA gene of B/Yamagata and B/Victoria lineages were designed by Primer Primer and then applied in the duplex real-time RT-PCR method that was followed developed. Influenza virus B type and A type isolated in our laboratory and typing-confirmed by HAI method were used as reference strains to determine the specificity of this assay and the sensitivity of the duplex amplification was evaluated by viral load testing in terms of in vitro transcribed RNA copy number.</p><p><b>RESULTS</b>In 2006-2010, 793 influenza virus type B strains were isolated from 17 765 throat swab samples, among which 152 strains were differentiated as By lineage and 641 as Bv lineage by this assay. These results was agreement with that determined by HAI assay. This developed assay allows to accurately identify approximately 10(2) copies/microl for Bv and By lineage virus with intra- and inter-coefficient of variation (CV) < 3.5% and nearly 100% specificity.</p><p><b>CONCLUSIONS</b>This method provides sensitive and robust tool for routine diagnosis and on-time epidemiological examination of influenza virus, which could be applied in influenza surveillance laboratories for rapid molecular diagnosis.</p>

Simultaneous detection of influenza virus type B and influenza A virus subtypes H1N1, H3N2, and H5N1 using multiplex real-time RT-PCR.

Use of multiplex real-time reverse transcription polymerase chain reaction (RT-PCR) for the simultaneous detection of influenza type B virus and influenza A virus subtypes H5N1, H3N2, and H1N1 has been described. The method exhibited a high specificity and sensitivity of approximately 10(1)-10(2) copies per microliter or 10(-3)-10(-2) TCID50/L for each subtype, as well as a high reproducibility with coefficient of variation (CV) ranging from 0.27% to 4.20%. The assays can be performed commendably on various models of real-time PCR instruments; including ABI7500, ROCH 2.0, and Mx3005p. In an analysis of 436 clinical samples from patients during the year 2009, this detection method has successfully identified 261 positive samples, as compared to only 189 positive samples using the conventional cell culture systems, and at the same time further differentiated them as 35 type B, 21 subtype H1N1, and 205 subtype H3N2. The results indicate that the multiplex real-time RT-PCR method is a potential tool for rapid screening of influenza virus from a large pool of clinical samples during flu pandemics and facilitates early influenza virus identification in most public health laboratories around the world.

Compare real-time RT-PCR with two culture methods for influenza virus detection / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Real-time RT-PCR, cell culture and embryonated eggs culture for influenza detection were compared by analyzing the data of influenza surveillance in Shenzhen in second half of 2009.</p><p><b>METHODS</b>1092 clinical samples (throat swabs) collected during second half of 2009 were tested by real-time RT-PCR, cell culture and embryonated eggs culture, and the results were analyzed by statistical methods.</p><p><b>RESULTS</b>The positive rate were 54.21%, 27.11% and 16.21% using real-time RT-PCR, cell culture and embryonated eggs culture, and the sensitive were 100%, 50% and 29.9%. The lowest dilutions of virus detected by real-time RT-PCR were 10(-2) TCID50/ml.</p><p><b>CONCLUSION</b>The sensitive of real-time RT-PCR was higher than culture and the specificity was also very high. It was more suitable for emergency detect. The sensitive of cell culture for H3N2 subtype was higher, and sensitive of embryonated eggs culture for type B was higher.</p>

Analyses of serological and genetic characteristics on novel H1N1 influenza A virus from the infected patient in Shenzhen / 病毒学报

Analysis of serological and genetic characteristics on 2009 swine-origin influenza A (H1N1) virus (S-OIV) isolated from four patients with severe disease in Shenzhen were performed. Microneutralization assay showed that the neutralizing antibody titers of the infected patients did not exceed 1 : 20 in a short term post infection, which could not neutralize the viruses efficiently. Hemagglutination inhibition (HI) tests confirmed that the antigenicity of S-OIV from the patients was distinct from the seasonal influenza A virus, but similar to the reference strains of S-OIV. Phylogenetic and molecular analysis showed that S-OIV from the patients still belonged to the classical swine lineages and did not have the genetic characteristics of highly pathogenic influenza virus. Several amino acid residue mutations on HA protein were detected, which seemed not to affect the virulence and pathogenicity of the viruses. Further, A His 275 Tyr mutation on NA protein of a virus strain was detected, which induced the oseltamivir resistance of the virus.

Laboratory diagnosis and molecular characterization analysis of the H5N1 influenza virus isolated from the first human case in Shenzhen, China / 病毒学报

The tracheal aspirates and serum samples of a suspected human case of high-pathogenic avian influenza (firstly found in Shenzhen, China) were collected and tested by a series of assays. The results showed that the RNA extracted from the tracheal aspirate specimens of the patient was confirmed positive for H5N1 avian influenza virus by Real-time PCR. The H5N1 avian influenza virus was isolated from patient's tracheal aspirates on MDCK cell and was named A/Guangdong/2/06(H5N1). The viral load of tracheal aspirates collected at different time points were detected by Real-time PCR. The virus microneutralization and the antigenic ratio of human H5N1 isolated were also assayed. It was found that when the virus load decreased gradually after the disease onset, the serum neutralizing antibody titer in the patient increased to 1 : 160 and subsequently decreased gradually. By molecular analysis, the eight gene segments of A/Guangdong/2/06 revealed to be similar to that of H5N1 avian influenza viruses isolated from south China in 2005-2006. However, there were obvious differences in the gene sequence of the detected H5N1 viral RNA as compared with that of the strains isolated from Vietnam, Thailand and Indonesia.

Study on molecular epidemiological characteristics of influenza H1N1 viruses circulating in Shenzhen,China from 2005 to 2007 / 中华流行病学杂志

Objective To study the genetic and epidemiological characteristics of HA1 of influenza H1N1 viruses circulating in Shenzhen from 2005 to 2007. Methods The HA1 region was analyzed by RT-PCR and subsequently sequenced to analyze the HA1 genetic evolution. Phylogenetic analysis was confirmed on the homology of nucleitide comparing with the reference viruses of vaccines recommended by WHO and representative virus confirmed by China CDC. Relationship between isolation rates and genetic evolutions was explored. Results The average isolation rate from 2005 to 2007 was 7.16%. Of the isolates, the proportions of influenza H1N1 viruses in 2005, 2006 and 2007 were 56.14%, 66.03%,3.61% ,respectively. Data from HA1 phylogenetic analysis showed that there were at least three clades circulated in Shenzhen. Different viruses isolated during January to April were clustered with A/New Caledonia/20/1999 viruses isolated in the latter months of 2005 clustered with A/Solomon Island/3/2006 and viruses from 2006 to 2007 were in the same clade with A/GDLH/219/2006. Results showed that most viruses had a deletion of lysine at position 130. Compared with A/New Caledonia/20/1999, the virus isolated after May of 2005 occurred T82K, Y94H, R146K, R209K, T267N amino acid substitution, while some virus isolated after May 2006 took place the amino acid substitutions of A190T, H193Y,E195D (located at antigenic site B) and R146K(antigenic site A). The sequences at the receptor-binding sites and glycosylation sites were conserved. Compared with referring viruses, A/SZ/68/2007 had 50 amino acid substitutions in the HA1 region.Of these,eleven and six were located at antigenic sites and receptor-binding sites,respectively.Four amino acid substitution resulted in the deletion of glycosylation site.Conclusion Three different genetic lineages of influenza H1N1 virus were circulated in the population in Shenzhen during 2005-2007.The special virus named A/SZ/68/2007 should be paid further attention on its antigenic and epidemiological characteristics.