Clinical feature changes of a COVID-19 patient from mild to critical condition and cardiopulmonary pathological results / 中华心血管病杂志

Objective: To analyse the clinical history, laboratory tests and pathological data of a patient who suffered from novel coronavirus pneumonia(COVID-19) and provide reference for the clinical treatment of similar cases. Methods: Data of clinical manifestation, laboratory examination, bronchoscopy, echocardiography and cardiopulmonary pathological results were retrospectively reviewed in a case of COVID-19 with rapid exacerbation from mild to critical condition. Results: This patient hospitalized at day 9 post 2019 novel coronavirus(2019-nCoV) infection, experienced progressive deterioration from mild to severe at day 12, severe to critical at day 18 and underwent extracorporeal membrane oxygenation(ECMO) and continuous renal replacement therapy(CRRT) as well as heart lung transplantation during day 28-45 post infection, and died at the second day post heart and lung transplantation. The patient had suffered from hypertension for 8 years. At the early stage of the disease, his symptoms were mild and the inflammatory indices increased and the lymphocyte count decreased continuously. The patient's condition exacerbated rapidly with multi-organ infections, and eventually developed pulmonary hemorrhage and consolidation, pulmonary hypertension, right heart failure, malignant ventricular arrhythmias, liver dysfunction, etc. His clinical manifestations could not be improved despite viral RNAs test results became negative. The patient underwent lung and heart transplantation and finally died of multi organ failure at the second day post lung and heart transplantation. Pathological examination indicated massive mucus, dark red secretions and blood clots in bronchus. The pathological changes were mainly diffused pulmonary hemorrhagic injuries and necrosis, fibrosis, small vessel disease with cardiac edema and lymphocyte infiltration. Conclusions: The clinical course of severe COVID-19 can exacerbate rapidly from mild to critical with lung, liver and heart injuries.

Unsatisfying antiviral therapeutic effect in patients with mother-to-child transmissed chronic hepatitis B virus infection: a prospective multi-center clinical study / 中华医学杂志(英文版)

BACKGROUND@#Few data are available regarding the progression of liver disease and therapeutic efficacy in chronic hepatitis B virus (HBV) carriers infected by mother-to-child transmission (MTCT). This study aimed to investigate these two aspects by comparing the adult chronic HBV carriers in MTCT group with those in horizontal transmission group.@*METHODS@#The 683 adult chronic HBV patients qualified for liver biopsy including 191 with MTCT and 492 with horizontal transmission entered the multi-center prospective study from October 2013 to May 2016. Biopsy results from 217 patients at baseline and 78 weeks post antiviral therapy were collected.@*RESULTS@#Patients infected by MTCT were more likely to have e antigen positive (68.6% vs. 58.2%, χ = -2.491, P = 0.012) than those with horizontal transmission. However, in patients with MTCT, levels of alkaline phosphatase (ALP) (P = 0.031), Fibroscan (P = 0.013), N-terminal propeptide of Type III procollagen (PIIINP) (P = 0.014), and Laminin (LN) (P = 0.006) were high, in contrast to the patients with horizontal transmission for whom the levels of albumin (ALB) (P = 0.041), matrix metalloproteinase-3 (MMP-3) (P = 0.001) were high. The 47.2% of patients with MTCT and 36.8% of those with horizontal transmission had significant liver fibrosis (P = 0.013). Following antiviral therapy for 78 weeks, 21.2% and 38.0% patients with MTCT and horizontal transmission acquired hepatitis B e antigen (HBeAg) clearance, respectively (P = 0.043), and the virological response rates were 54.7% and 74.1% in the MTCT and horizontal groups, respectively (P = 0.005). MTCT was a risk factor for HBeAg clearance and virological response.@*CONCLUSION@#Adult patients with MTCT were more prone to severe liver diseases, and the therapeutic efficacy was relatively poor, which underlined the importance of earlier, long-term treatment and interrupting perinatal transmission.@*TRIAL REGISTRATION@#NCT01962155; https://clinicaltrials.gov.

Unsatisfying antiviral therapeutic effect in patients with mother-tochild transmissed chronic hepatitis B virus infection: a prospective multi-center clinical study / 中华医学杂志(英文版)

Background@#Few data are available regarding the progression of liver disease and therapeutic efficacy in chronic hepatitis B virus (HBV) carriers infected by mother-to-child transmission (MTCT). This study aimed to investigate these two aspects by comparing the adult chronic HBV carriers in MTCT group with those in horizontal transmission group.@*Methods@#The 683 adult chronic HBV patients qualified for liver biopsy including 191 with MTCT and 492 with horizontal transmission entered the multi-center prospective study from October 2013 to May 2016. Biopsy results from 217 patients at baseline and 78 weeks post antiviral therapy were collected.@*Results@#Patients infected by MTCT were more likely to have e antigen positive (68.6% vs. 58.2%, χ2 = -2.491, P = 0.012) than those with horizontal transmission. However, in patients with MTCT, levels of alkaline phosphatase (ALP) (P = 0.031), Fibroscan (P = 0.013), N-terminal propeptide of Type III procollagen (PIIINP) (P = 0.014), and Laminin (LN) (P = 0.006) were high, in contrast to the patients with horizontal transmission for whom the levels of albumin (ALB) (P = 0.041), matrix metalloproteinase-3 (MMP-3) (P = 0.001) were high. The 47.2% of patients with MTCT and 36.8% of those with horizontal transmission had significant liver fibrosis (P = 0.013). Following antiviral therapy for 78 weeks, 21.2% and 38.0% patients with MTCT and horizontal transmission acquired hepatitis B e antigen (HBeAg) clearance, respectively (P = 0.043), and the virological response rates were 54.7% and 74.1% in the MTCT and horizontal groups, respectively (P = 0.005). MTCT was a risk factor for HBeAg clearance and virological response.@*Conclusion@#Adult patients with MTCT were more prone to severe liver diseases, and the therapeutic efficacy was relatively poor, which underlined the importance of earlier, long-term treatment and interrupting perinatal transmission.@*Trial Registration@#NCT01962155; https://clinicaltrials.gov.

Clinical Observation of Pulmonary Rehabilitation Therapy for Patients with Severe and Extremely Severe Chronic Obstructive Pulmonary Disea / 中山大学学报(医学科学版)

@#【Objective】To explore the effect of pulmonary rehabilitation on patients with severe and extremely severe chronic obstructive pulmonary disease（COPD）. 【Methods】68 patients with chronic obstructive pulmonary disease admitted to Chaozhou people′s hospital from January 2014 to January 2016 were enrolled in the study，including 51 males and 17 females ，aged（64±8） years old. All patients were divided into treatment group（34 cases） and control group（34 cases）by random number. All patients received routine medication and life style guidance，while pulmonary rehabilitation was added to the treatment group. Follow up for 12 months ，and the data of lung function ，6 minute walking test（6MWT），COPD assessment test（CAT）scores and hospital anxiety and depression scale（HADs）scores were collected regularly.【Results】No statistically significant difference was found between the two groups in forced expiratory volume in one second/prediction（FEV1/Pre），FEV1/FVC，6MWT and HADs score after six months′ follow-up（P > 0.05），while the CAT scores of treatment group was superior to the control group（P < 0.05）. No statistically difference was found between the two groups in FEV1/Pre and HADs scores after 12 months′ follow-up（P > 0.05），while FEV1/FVC，6MWT and CAT scores of treatment group were superior to control group（P < 0.05，P < 0.05，P < 0.01，respectively）. The 6MWT and CAT scores of treatment group were superior to pretreatment after 6 months′ follow-up（P < 0.01，P < 0.001）and 12 months′ follow-up（P < 0.001，P < 0.001）.【Conclusion】Pulmonary rehabilitation for patients with severe and extremely severe COPD can effectively improve their symptoms，lung function，quality of life and exercise endurance.

Retinal hemodynamic influence of compound xueshuantong capsule on nonproliferative diabetic retinopathy after laser photocoagulation / 国际眼科杂志(Guoji Yanke Zazhi)

AlM: To observe retinal hemodynamic influence of compound xueshuantong capsule on nonproliferative diabetic retinopathy ( NPDR) after laser photocoagulation.METHODS:A total of 41 patients (72 eyes) with NPDR after laser photocoagulation were enrolled in this study. They were all given compound xueshuantong capsule, and used color Doppler flow imaging for detection of retinal hemodynamics. RESULTS: After treatment, patients with retinal blood perfusion significantly improved; central retinal arterial peak systolic velocity ( PSV ) , end - diastolic velocity (EDV) and medial velocity (Vm) were increased, while the resistance index ( Rl) decreased. The difference have statistical significance (P CONCLUSlON: Compound xueshuantong capsule can improve retinal blood perfusion for nonproliferative diabetic retinopathy after laser photocoagulation, which is related to improvement of visual prognosis.

Efficacy of Peg-interferon alpha-2a combinated with entecavir on HBeAg positive chronic hepatitis B patients with high serum hepatitis B viral loads / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>This study aimed at evaluating the efficacy and safety of a combination treatment of entecavir and Peginterferon alpha-2a for HBeAg positive chronic hepatitis B patients with high serum hepatitis B viral loads.</p><p><b>METHODS</b>60 treatment-naive HBeAg-positive CHB patients with high serum hepatitis B viral loads were enrolled and randomly divided into three groups: group A received Peginterferon alpha-2a monotherapy for 48 weeks (n = 20); group B received entecavir monotherapy for more than 48 weeks (n = 20); group C received Peginterferona alpha-2a combined with entecavir for 12 weeks, then Peginterferon alpha-2a monotherapy for 36 weeks (n = 20). Virological response, ALT normalization, HBeAg and HBsAg seroclearance rate were analysed at the end of 4, 12 and 24 weeks after the treatment.</p><p><b>RESULTS</b>The ratio of undetectable hepatitis B virus (HBV) DNA were 50% and 10%, 95% and 25% and 100% and 30% in group C and group A respectively, 50% and 20%, 95% and 75% and 100% and 90% in group C and group B respectively at the end of 4, 12 and 24 weeks of treatment. The differences were significant between group C and A (Z = -4.6, P < 0.001), group C and B (Z = -2.53, P = 0.0114). ALT normalization rate was significantly lower in group A than that of group C (Z = -2.63, P = 0.0086). HBeAg levels declined more in group C than the other two groups after 24 weeks of treatment.</p><p><b>CONCLUSIONS</b>For HBeAg positive chronic hepatitis B patients with high serum hepatitis B viral loads, combination treament of Peginterferon alpha-2a with entecavir is more effective than Peginterferon alpha-2a monotherapy in virologic response and ALT normalization after 24 weeks of treatment.</p>

Soluble expression of A/H1N1 influenza virus HA with Drosophila S2 cell line and its bio-activity identification / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To express soluble HA of A/H1N1 influenza virus in drosophila S2 cell line and identify its bio-activity.</p><p><b>METHODS</b>HA gene was amplified from A/Shenzhen/71/09 virus strain using RT-PCR, then we constructed pAC5.1-HA expression vector, which was co-transfected into S2 cell with pCoblast vector. After transfection, stable S2 cell was selected through Blasticindin. HA in the supernatant was identified with Western Blot assay and purified with Ni-column. Recombinant HA was immunized into BALB/c mice 3 times, and the Abs titers were evaluated with ELISA.</p><p><b>RESULTS</b>We successfully cloned HA gene with 1.7 x 10(3) bp of A/Shenzhen/71/09 virus strain and got recombinant pAC5. 1-HA expression vector. Stable S2 cell line was established after transfection and selection, which continuously expressed HA with molecular weight 75 x 10(3) D. After immunization with HA, the Abs titers were 1:1280 and 1: 5120 respectively on 10 d, 30 d.</p><p><b>CONCLUSION</b>We expressed soluble HA with good bio-activity, which contributed to research on immune diagnosis, subunit vaccine, and monoclonal Abs for influenza.</p>

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>

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>

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 association between HBV genotyping and clinical characteristics and expression of TH1/TH2 cytokines / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore the association between HBV genotyping and clinical characteristics and expression of TH1/TH2 cytokines.</p><p><b>METHODS</b>The expression of IL-4 and IFN-gamma was detected with flow cytometry for 102 HBV infections and 48 healthy controls. 50 CHB patients were randomly selected for HBV genotyping with real-time fluorescence PCR assay.</p><p><b>RESULTS</b>Higher expression of IL-4 in peripheral blood was detected in patients with HBV infection than healthy controls (P < 0.001); No significant differences on expression of Th1/Th2 cytokines were observed in CHB patients with different HBV DNA levels or HBeAg status (P > 0.05). There were 34 (68%) patients with genotype B infection and 16 (32%) with genotype C infection. Compared to patients with genotype B infection, the patients with genotype C infection showed higher levels of IL-4 (P = 0.018), and Th1/Th2 ratio decreased,but the difference was not statistically significant (P = 0.2262).</p><p><b>CONCLUSION</b>The different expression of TH1/TH2 cytokines may elucidate cellular immune response and clinical outcome difference between patients with genotype B infection and genotype C infection.</p>

Loss of the balance between Th17 and Th1 populations in HIV/AIDS patients / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To evaluate the Th17/Th1 response in HIV infected patients and the mutual relationship between the response of Th17 and Th1.</p><p><b>METHODS</b>38 chronic HIV infected patients as well as 24 healthy volunteers were performed in this study. The patients were divided into two groups, one group before treatment, the other after therapy. The whole blood intracellular cytokine staining was used, samples detected by BD FACSCanto, after that, the expression of CD4+ IL-17+ T cell and CD4 IFN-gamma+ T cell were analyzed by FACSDiva software and lastly compared the differences among different groups.</p><p><b>RESULTS</b>The expression of CD4+ IL-17+ T cell in naive-therapy patients were significantly lower than that of the healthy controls (1.14 +/- 0.7)9% vs (3.98 +/- 1.14)%, P = 0.000, but increased remarkably after HARRT(highly antiretroviral treatment) (2.22 +/- 1.00)%, P = 0.001; however there were no significant differences in the expression of CD4+ IFN-gamma+ T cell before and after therapy (34.35 +/- 24.38)% vs (42.10 +/- 15.57%), also with the healthy control (P = 0.383). The frequency of CD4 IL-17+ T cell was positively correlated with CD4+ T counts (R = 0.345, P = 0.034), but no significant correlations was observed between the expression of CD4+ IFN-gamma T cell and CD4+ T counts (R = -0.247, P = 0.136).</p><p><b>CONCLUSION</b>The infection of HIV virus down-regulated Th17 immune response and disturbed the balances between Th17 and Th1 evidently in human. Th17 response may play an important role in the pathogenesis of HIV infection.</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>

The expression of some homing and co-receptor molecules on CD4+ T cells in AIDS patients / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To evaluate the expression of CD49d, CCR9, CD62L and CCR5 on CD4+ T cells in AIDS patients before and after HAART.</p><p><b>METHODS</b>The study was performed in 42 cases of AIDS patients and 18 cases of healthy controls. The expression of CD49d, CCR9, CD62L and CCR5 on CD4+ T cells, and CD45RO on CD4+ CCCR9+ and CD4+ CCR5+ T cells in AIDS patients and healthy controls were analysed by Flow cytometry. Software BD FACSDiva was used to calculate the percentage of expression.</p><p><b>RESULTS</b>The number of peripheral CD4+ T cells in group pre-HAART was decreased compared with group HAART (P < 0.01); the frequency of CD3+ CD4+, CD4+ CCR9+, CD4+ CCR5+ T cells in group pre-HAART were decreased compared with group HAART (P < 0.01), the frequency of CD4+ CD49d+, CD4+ CD62L+, CD4+ CCR9+ CD45RO , CD4+ CCR9+ CD45RO- ,CD4+ CCR5+ CD45RO+, CD4+ CCR5+ CD45RO- T cells in group pre-HAART were significantly decreased compared with group HAART (P < 0.001); the frequency of CD3+ CD4, CD4+ CD62L+, CD4+ CCR5+ T cells in group HAART were significantly decreased compared with group HIV-neg (P < 0.05).</p><p><b>CONCLUSIONS</b>Not only the number but the function of CD4' T cells was impaired in AIDS patients: the lower expression frequency of gut homing receptor molecule CD49d and CCR9,1ymph node homing molecule CD62L, coreceptor molecule CCR5. HAART can partially reverse this pathological phenomena. CD49d, CCR9 and CD62L may be suggested to indicate the progression of AIDS and immunologic reconstitution after HAART.</p>

Effect of hepatitis B virus C protein on function of natural killer cell in NK-92 cells / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the influence of hepatitis B virus C protein on the function of natural killer cell.</p><p><b>METHODS</b>Recombinant eukaryotic expression plasmid pHBI-CMV-HBC was constructed and confirmed by double restrictive enzyme digestion and DNA sequencing analysis. Then the recombinant plasmid was transfected into NK-92 cells with lipofectamine encapsuled. The transfected NK-92 cells containing expressive HBV C protein was confirmed by Western Blot analysis. ELISA was employed to determine the IFN-gamma level secreted by NK-92 cells. And finally the cytotoxicities of NK cells were analysed by MTT colorimetry, with the hepatoblastoma cell line (HepG2) as target cell.</p><p><b>RESULTS</b>Western blotting confirmed the expression of HBV C protein in the NK-92 cells transfected with pHBI-CMV-HBC. NK cytotoxicities and IFN-gamma secretion level of NK-92 cells transfected with recombinant plasmid significantly increased compared to control NK-92 cells transfected with blank plasmid (P < 0.01) and untransfected NK-92 cells(P < 0.01).</p><p><b>CONCLUSION</b>Transient expression of HBC can increase IFN-gamma secretion and cytotoxicities of NK-92 cells.</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>

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>

Dynamics of in vitro amyloid fiber formation of yeast prion protein Sup35NM / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To investigate the dynamics of amyloid fiber formation of yeast (Saccharomyces cerevisiae) prion protein Sup35NM under the native condition to provide materials and clues for the elucidation of amyloid fiber formation.</p><p><b>METHODS</b>The Sup35NM gene was cloned and expressed in E. coli. The recombinant Sup35NM protein was purified under denaturing conditions through Nickel-Sepharose chromatography. Aliquots were removed at designated time points for transmission electron microscopy (TEM), circular dichroism (CD) spectra, protease K resistance assay, as well as thioflavin T (ThT) binding assay.</p><p><b>RESULTS</b>The Sup35NM expressed and purified under denaturing conditions. The morphological alteration of the Sup35NM in PBS (pH7.4) during the protein aggregation and amyloid fiber formation was visualized by TEM. The CD assay showed that the course of amyloid fiber formation underwent a conformational shift from alpha-helix to beta-sheet. The fibers had higher capacity of resistance to protease K digestion compared to the monomers. ThT fluorescence assay displayed a rapid growth phase before reaching a final equilibrium phase during the fiber formation, and the higher concentration of Sup35NM could greatly accelerate the fiber formation in vitro.</p><p><b>CONCLUSION</b>Yeast prion protein Sup35NM forms amyloid readily under native conditions in vitro. The dynamics of Sup35NM amyloid formation may provide supporting evidences for the nucleating polymerization models of amyloid fiber formation.</p>

Gene expression profile of peripheral blood monocyte in patients with fulminant hepatitis B by cDNA microarray / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate peripheral blood monocyte (PBMC) gene expression profile in patients with fulminant hepatic failure (FHF) by cDNA microarray.</p><p><b>METHODS</b>Microarrays consisting of 8,192 human cDNAs and labelled cDNAs prepared from PBMC in both 10 FHF patients and 10 asymtomatic surface antigen carriers (ASC) were applied to analyze gene expression. Relative ratios of gene expression in individuals were obtained by comparing the hybridization results, by GenePix 4000B scanning and by ImaGene3.0 software analysis, of Cy5-labelled cDNA from FHF patients with those of Cy3-labelled cDNA from ASC.</p><p><b>RESULTS</b>249 genes out of 8,192 were identified differently, at least two times. Most of the genes (79%) involved in cell signaling transduction, cell cycles, metabolism, inflammatory response and apoptosis, whose mRNAs were differently regulated.</p><p><b>CONCLUSIONS</b>These results suggest that HBV infection alters a broad range of cellular genes expression during developing into FHF and provide a framework for future functional study on the genes expressed differently.</p>