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Yirui Xie,1 Bohao Dai,1 Xiaotang Zhou,1 Huiting Liu,1 Wei Wu,2 Fei Yu,3 Biao Zhu1 1State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The Department of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, 310003, People’s Republic of China; 2State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, 310003, People’s Republic of China; 3Key Laboratory of Clinical in vitro Diagnostic Techniques of Zhejiang Province, Department of Laboratory Medicine, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310003, People’s Republic of ChinaCorrespondence: Yirui Xie; Biao Zhu, State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The Department of Infectious Diseases, The First Affiliated Hospital, School of Medicine, Zhejiang University, 79, QingChun Road, Hangzhou, 310003, People’s Republic of China, Tel/Fax +86-571-87236416, Email 1312019@zju.edu.cn; zhubiao1207@zju.edu.cnBackground: To evaluate the value and challenges of real-world clinical application of metagenomic next-generation sequencing (mNGS) for bronchoalveolar lavage fluid (BALF) in HIV-infected patients with suspected multi-pathogenic pneumonia.Methods: Fifty-seven HIV-infected patients with suspected mixed pneumonia who were agreed to undergo the bronchoscopy were recruited and retrospectively reviewed the results of mNGS and conventional microbiological tests (CMTs) of BALF from July 2020 to June 2022.Results: 54 patients were diagnosed with pneumonia including 49 patients with definite pathogens and five patients with probable pathogens. mNGS exhibited a higher diagnostic accuracy for fungal detection than CMTs in HIV-infected patients with suspected pulmonary infection. The sensitivity of mNGS in diagnosis of pneumonia in HIV-infected patients was much higher than that of CMTs (79.6% vs 61.1%; P < 0.05). Patients with mixed infection had lower CD4 T-cell count and higher symptom duration before admitting to the hospital than those with single infection. The detection rate of mNGS for mixed infection was significantly higher than that of CMTs and more co-pathogens could be identified by mNGS. The most common pattern of mixed infection observed was fungi-virus (11/29, 37.9%), followed by fungi-virus-bacteria (6/29, 20.7%) coinfection in HIV-infected patients with multi-pathogenic pneumonia.Conclusion: mNGS improved the pathogens detection rate and exhibited advantages in identifying multi-pathogenic pneumonia in HIV-infected patients. Early performance of bronchoscopy and mNGS are recommended in HIV-infected patients with low CD4 T cell counts and long duration of symptoms. The most common pattern of mixed infection observed was fungi-virus, followed by fungi-virus-bacteria coinfection in HIV infected patients with multi-pathogenic pneumonia.Keywords: HIV, multi-pathogenic pneumonia, metagenomic next-generation sequencing, conventional microbiological tests, bronchoalveolar lavage fluid

Aims To study the role and characteristics of activated microglia in poststroke depression (PSD) . Methods Twenty-four male Wistar rats were randomly divided into three groups: the poststroke (PS) group, PSD group, and Sham group. Neurobehavioral testing was performed 24 h postoperation. The body weights of the rats were regularly recorded, and behavioral testing was regularly performed at 1, 2 and 3 weeks postmodeling. Immunofluorescent staining was used to detect the microglial marker OX42. Real-time PCR was used to analyze the relative gene expression of microglial activation markers (TNF-a, IL-10, IL-1, TGF-β, CD86, iNOS, CD206, IL-1β, and Arg1) . Results The relative gene expression of proinflammatory markers (IL-1, TNF-a, iNOS, and IL1β) and anti-inflammatory markers (CD206 and Arg1) significantly increased in the hippocampal region compared with that in the right cerebral and left cerebral hemispheres in the PSD group. The relative gene expression of proinflammatory markers (TNF-a, IL-1, iNOS, and CD86) in the hippocampal region was significantly increased in the PSD group compared with that in the Sham and PS groups. The anti-inflammatory markers (TGF-β and CD206) in the hippocampal region were significantly increased in the PSD group compared with that in the Sham group, and the M2 marker Arg1 was significantly increased in the PSD group compared with that in the PS group. Correlation analysis showed that IL-1 was strongly negatively correlated with PSD . Conclusions Most microglia in the hippocampal region of PSD had a proinflammatory status and an anti-inflammatory status. IL-1 showed a strong negative correlation with PSD.

Background T cells play an important role in the prognosis of hepatitis B virus (HBV) infection, and are involved in the seroconversion of a patient from HBsAb negative to positive. To compare the T-cell receptor β-chain variable region (TcRBV) complementarity-determining region 3 (CDR3) in subjects with or without hepatitis B surface antigen (HBsAg) convert to hepatitis B surface antibody (HBsAb), the TcRBV was determined using high throughput sequencing (HTS). Methods The clonotype and diversity of CDR3 in peripheral blood mononuclear cells of subjects with resolved acute hepatitis B (AHB, HBsAb+, HBsAg-) (n = 5), chronic hepatitis B (CHB, HBsAb-, HBsAg+) (n = 5), and healthy controls (HC, HBsAb-, HBsAg-) (n = 3) were determined and analyzed using HTS (MiSeq). Results The overlapping rate of CDR3 clones of any two samples in AHB group was 2.00% (1.74% ~ 2.30%), CHB group was 1.77% (1.43% ~ 2.61%), and HC group was 1.82% (1.62% ~ 2.12%), and there was no significant difference among the three groups by Kruskal-Wallis H test. However, among the top 10 cumulative frequencies of clonotypes, only the frequency of clonotype (TcRBV20–1/BD1/BJ1–2) in AHB group was lower than that of HC group (P P > 0.05). Conclusions Thus, there are 57 TcRBV clonotypes that may be related to HBsAg seroconversion of AHB subjects, but the diversity of TcRBV CDR3 is not significantly related to the HBsAb positive status.

This study tests for evidence of tonal coarticulation effects, especially anticipatory effects, in production of non-native Chinese contour tones. Eighty second language learners of Chinese and ten native speakers participated in a main experiment and two supplementary experiments in which they produced both real and pseudo disyllabic words. Findings indicate that anticipatory coarticulation is relevant in L2 contour tone production. L2 speakers’ Tone 2 and to some extent Tone 4 tend to be less intelligible to native listeners when followed by tones starting with a high onset (Tone 1 or Tone 4) due to anticipatory coarticulation. Some similar and different tonal coarticulation effects between native Chinese speakers and second language learners of Chinese are also noted in the paper based on the experiment results. This study argues that a ‘universal’ coarticulatory constraint plays a role in shaping second language Chinese tone phonology.

Dielectric barrier discharge (DBD) plasma coupled with Fe–Mn doped AC (Fe–Mn/AC) was used to enhance the degradation of tetracycline hydrochloride (TCH) wastewater. Fe–Mn/AC catalysts with different Fe/Mn molar ratios were prepared by hydrothermal method, and the physical and chemical properties of the samples were explored by different characterization techniques, including XRD, SEM, TEM and XPS. The results showed that the combination of DBD with Fe2–Mn1/AC system had the highest effect, and the degradation efficiency of TCH could reach 98.8 % after 15 min treatment, which was 25.5 % higher than that of DBD-only. With the increase of discharge voltage and catalyst dosage, the degradation efficiency of TCH promoted. And initial pH had little effect on the degradation of TCH. In the combined system, the Fe2–Mn1/AC catalyst could retain an excellent stability and reusability. The addition of dimethyl sulfoxide (DMSO) showed that ·OH participated in the TCH degradation. The generated O3 might be catalyzed by Fe–Mn/AC catalyst to produce more ·OH. And more H2O2 was produced in DBD-only system than that in DBD-catalytic system. Nine main degradation intermediate products in the combined system were detected by HPLC-MS, and three possible degradation pathways were proposed.

We report here a novel HIV-1 recombinant form detected from a married HIV-positive man infected through homosexual behavior in Zhejiang, China. The breakpoint analysis of near full-length genome demonstrated a complex genome organization comprising two circulating recombinant forms (CRFs), CRF01_AE and CRF07_BC, both well-known CRFs in China. The parental CRF01_AE region (II) of recombinant clustered together with a previously reported cluster 4 lineage. The CRF07_BC regions (I and III) clustered within CRF07_BC references. The ongoing generation of intersubtype recombinant viruses increases the complexity of the HIV-1 epidemic and illustrates the necessity of persistent surveillance of the transmission of HIV-1.

Additional file 2: Figure S1. Frequency of V/J gene combinations have significant differences among the three groups (P

Additional file 1: Table S1. The information of amplification. Table S2. Clonal overlap rate (BUB index) among the three groups.

Background: Although gut microbiota dysbiosis has recently been reported in HIV infected individuals, the relationship between the gut microbiota and immune activation in patients with a different immune response to highly active antiretroviral therapy (HAART) is still not well understood. Gut microbiota and immune activation were studied in 36 non-HIV-infected subjects (Health Control) and 58 HIV-infected individuals which included 28 immunological responders (IR) and 30 immunological non-responders (INR) (≥500 and Fusobacterium, Ruminococcus_gnavus and Megamonas were more abundant, whereas Faecalibacterium, Alistipes, Bifidobacterium, Eubacteriumrectale and Roseburia were more depleted in the IR and INR groups than that in the health control. Ruminococcaceae and Alistipes were positively correlated with nadir and current CD4+ T-cell counts, but negatively correlated with CD8+CD57+ T-cell counts. Inflammation markers and translocation biomarkers (LPS) levels were positively correlated with the abundance of genus Ruminococcus and Fusobacterium, but were negatively correlated with the genus Faecalibacterium. The relative abundances of Escherichia-Shigella and Blautia were significantly higher in the IR than that in the INR group. Escherichia-Shigella were negatively correlated with the CD4/CD8 ratio but positively correlated with the amount of CD8+CD57+ T-cells. Roseburia and Blautia were negatively associated with nadir CD4+ T-cell and positively associated with CD8+CD57+ T-cell counts. Conclusions: Gut microbiota dysbiosis may be one of the factors contributing to different immune response to HAART. Fusobacterium, Alistipes, Ruminococcaceae, Faecalibacterium, Escherichia-Shigella, Roseburia and Blautia maybe the major genus contributed to different immune response in immunodiscordant and immunoconcordant patients on long-term suppressive ART. Blautia and Roseburia might be associated with treatment outcome.

The hierarchical microtextured/nanotextured topographies have been recognized to have better tissue integration properties, but the underlying mechanisms are only partially understood. Hedgehog signaling plays a pivotal role in developmental and homeostatic angiogenesis. We suppose that the Hedgehog-Gli1 signaling may play a significant role in the response of endothelial cells to microtextured/nanotextured topographies (MNTs). To confirm this hypothesis, we produced the MNTs decorated with TiO2 nanotubes of two different diameters (25 and 70 nm), and the proliferation, apoptosis, angiogenesis-related genes expression and Hedgehog signaling activity of human umbilical vein endothelial cells (HUVECs) grown onto these MNTs were measured. Our results showed that the MNTs induced significantly high expression of Sonic Hedgehog (SHH), Smoothened (SMO) and GLI1 in the HUVECs as well as high activation of Hedgehog-Gli1 signaling, compared to the smooth surface. The HUVECs grown on the MNTs showed significantly high levels of adhesion, proliferation and expression of angiogenesis-related genes, including angiopoietin-1 (ANG-1), vascular endothelial growth factor (VEGFA), vascular endothelial growth factor receptor 2 (VEGFR2) and endothelial nitric oxide synthase (ENOS); these enhancements were attenuated by siRNA-mediated depletion of SMO, which indicated a significant role of Hedgehog-Gli1 signaling in mediating the enhanced effect of the MNTs on the angiogenic potential of HUVECs. This study may contribute to the modification of biomaterial surfaces for better tissue integration and clinical performance.

Micro/nanotextured topographies (MNTs) can modulate cell-biomaterial interactions mostly by their controllable geometrics. Among them, TiO2 nanotubes, regarded as having a highly controllable nanoscale geometry, have been extensively investigated and applied and significantly affect diameter-dependent cell biological behaviors. In this study, we used five typical MNTs decorated with TiO2 nanotubes with diameters of 30, 50, 70, 100 and 120 nm to explore the optimal nanotube diameter for improving the biofunctional properties and to more deeply understand the underlying mechanisms by which these MNTs affect osteogenic differentiation by revealing the effect of beta1-integrin/Hedgehog-Gli1 signaling on this process. The MNTs affected MG63 osteoblast-like cell spreading, osteogenic gene expression (BMP-2, Runx2 and ALP), mineralization and ALP activity in a diameter-dependent pattern, and the optimal TiO2 nanotube diameter of 70 nm provided the best microenvironment for osteogenic differentiation as well as beta1-integrin/Hedgehog-Gli1 signaling activation. This enhanced osteogenic differentiation by the optimal-diameter TiO2 nanotubes of 70 nm was attenuated via suppression of the beta1-integrin/ Hedgehog-Gli1 signaling, which indicated a significant role of this pathway in mediating the diameter-dependent osteogenic differentiation promotional effect of MNTs with different TiO2 nanotube diameters. These results might provide deeper insights into the signal transduction mechanisms by which different nanoscale geometries influence cellular functions for biomaterial modification and biofunctionalization.

Introduction Human papillomavirus (HPV) vaccination has been proven to effectively protect against HPV infection and infection-associated cancer. However, there are concerns about the relationship between HPV vaccination and the risk of autoimmune disorders (ADs). Therefore, we performed a systematic review and meta-analysis to comprehensively evaluate the relationship between HPV vaccination and ADs risk. Methods To identify relevant studies, we conducted a systematic search in EMBASE and PubMed databases of scientific articles published through June 2018. Fixed or random effects models were adopted to estimate overall relative risk. Results In total, 20 studies (12 cohort studies, 6 case-control studies, and 2 randomized controlled trials) involving more than 169,000 AD events were included in our meta-analysis. Our results show that HPV vaccination was not associated with an increased risk of subsequent ADs (odds ratio [OR] = 1.003, 95% confidence interval (CI): 0.95–1.06), particularly among those with a prior ADs (OR = 0.82, 95% CI: 0.7–0.96). Most of the subgroup analysis results based on the location or type of ADs were consistent with the overall results. Conclusion No evidence of an association between HPV vaccination and ADs was found. Given the low number of estimates for individual AD, additional and larger observational studies are needed to verify our findings.

The relationship between hemostatic system and HBeAg seroconversion (SC) of chronic hepatitis B (CHB) patients is ill-defined. We therefore evaluate the predictive value of plasma ADAMTS13 (a disintegrin and metalloproteinase with a thrombospondin motif repeats 13) and VWF (von Willebrand factor) for CHB patients during 5-year entecavir (ETV) treatment. One hundred and fourteen HBeAg positive CHB patients on continuous ETV treatment were recruited. Liver biopsies were evaluated using the METAVIR scoring system, and plasma ADAMTS13 activity (ADAMTS13: AC) and VWF antigen (VWF: Ag) were determined at baseline, 3, 12, 24, 36, and 60 months, respectively. ETV treatment resulted in an increased ADAMTS13: AC and decreased VWF: Ag (both P P

Objective: We aimed to analyze alterations in T cell subgroups during different post-ischemic stroke (IS) phases to explore the possible mechanisms underlying stroke-induced immune depression (SIID). Methods: Sixty-four IS patients who met the entry criteria were divided into three groups: an acute phase group, a sub-acute phase group and a stable phase group. Fourteen healthy individuals were selected as normal controls. The phenotype distribution of T cells in patient peripheral blood was analyzed, and the immune checkpoint receptors programed cell death-1 (PD-1) and T cell immunoglobulin and mucin domain 3 (Tim-3) were detected in different T cell phenotypes. Results: Compared with the control group, the absolute number of CD4+ T cells and CD4+ T central memory (TCM) cells was significantly increased in the acute phase group but decreased in the sub-acute phase and stable phase groups compared with that in the acute phase group. PD-1 expression in CD4+ T cells in the stable phase group showed a significant increase compared with that in the acute phase group. The expression of PD-1 on CD4+ TCM cells and CD4+ T effector memory (TEM) cells showed significant decreases in the acute phase compared with control cells; however, in the sub-acute phase and the stable phase, PD-1 expression was significantly increased compared with that in the acute phase. Conclusions: T cell dysfunction, especially CD4+ T cell dysfunction, occurred during different IS phases. PD-1 was highly expressed in CD4+ T cells of different phenotypes after the acute phase and was associated with alterations in CD4+ T cells. Particularly, PD-1 was negatively correlated with the absolute number of TCM cells among different CD4+ T cell phenotypes, which may be one of the possible mechanisms of SIID.

Background. The etiology of immune reconstitution inflammatory syndrome (IRIS) in AIDS patients after the initiation of HAART remains unknown. Several researches indicated that the development of IRIS is associated with the production and variation of cytokines, whose gene expression are closely related to the Ca2+/CN-nuclear factor of activated T cells (NFAT) pathway.Methods. We studied the expression of NFAT isoforms and their major target cytokines genes in peripheral blood CD3+T cells of subjects through fluorescence quantitative PCR and explored the expression changes of these genes before and after HAART.Results. After the initiation of HARRT, NFAT1, IL-6, and IL-8 gene expression showed a reversal trend in the CD3+T cells of the IRIS group and changed from low expression before HARRT to high expression after HARRT. In particular, the relative gene expression of NFAT1 was markedly higher compared with the other three isoforms. The IRIS group also showed higher NFAT4, NFAT2, NFAT1, IL-1β, IL-10, IL-2, IL-18, and TNF-αgene expression than the non-IRIS group.Conclusion. This study suggested that high expression levels of IL-2, IL-6, IL-8, TNF-α, IL-1β, IL-10, IL-12, and IL-18 can predict the risk of IRIS. The increased expression of NFAT1 and NFAT4 may promote the expression of cytokines, such as IL-6, IL-8, and TNF-α, which may promote the occurrence of IRIS.

The profile of T-cell receptor beta-chain variable (TRBV) genes usually skews in subjects with virus infection or cancer. The gene melting spectral pattern (GMSP) can be used to determine the profile of the TRBV gene family. To explore the portrait of the TRBV family in peripheral blood lymphocytes from subjects who have recovered from acute hepatitis B virus infection (AHI), peripheral blood mononuclear cells (PBMCs) were separated and further sorted into CD4+ and CD8+ T-cell subsets. The molecular features of the TRBV complementary determining region 3 (CDR3) motifs were determined using GMSP analysis. When a GMSP profile showed a single peak, the monoclonally expanded TRBV gene was cloned and sequenced. Skewed expansions of multiple TRBV genes were observed among the CD4+ and CD8+ T-cell subsets and the PBMCs. The frequency of monoclonally expanded TRBV genes in the CD8+ T-cell subset was significantly higher than that of the CD4+ T-cell subset and the PBMCs. Compared to other members of the TRBV gene family, TRBV11, BV15 and BV20 were predominantly expressed in the repertoire of peripheral blood lymphocytes in recovered AHI subjects. The relatively conserved amino acid motifs of TRBV5.1 and BV20 CDR3 were also detected in the CD4+ and CD8+ T-cell subsets. These results demonstrate the presence of multiple biased TRBV families in recovered AHI subjects. TRBV11, BV15 and BV20, especially from the CD8+ T-cell subset, may be relevant to the pathogenesis of subjects with AHI. The preferentially selected TRBV5.1 and BV20 with the relatively conserved CDR3 motif may be potential targets for personalized treatments of chronic HBV infection.

Objective. To identify blood biomarkers to predict severity and mortality in AIDS PCP patients.Methods. Biomarkers including clinical parameters and plasma inflammatory cytokines were assessed in 32 HIV-infected patients withPneumocystispneumonia (PCP) at time of admission. Predictive value of the biomarkers for clinical severity and in-hospital mortality was evaluated by corresponding ROC curve.Results. Levels of CRP, WBC, LDH, HBDH, and Ferritin were significantly higher in the severe and nonsurvivor AIDS PCP patients. These important biochemical indicators have inverse correlation with oxygenation index, especially levels of LDH (P=0.008,R2=0.258), HBDH (P=0.001,R2=0.335), and Ferritin (P=0.005,R2=0.237). Plasma IL-8 and IL-6 levels were significantly higher in patients with PaO2/FiO2≤ 200 mmHg and nonsurvivors than in those with PaO2/FiO2> 200 mmHg and survivors. Severe and nonsurvival groups showed higher ratio of mean IL-6/IL-10 level (1.78 ± 1.56,P<0.001; 1.11 ± 0.72,P=0.043), larger AUC (95% CI 0.781–1.000,P<0.001; 95% CI 0.592–0.917,P=0.043), and more significantly inverse correlation with the oxygenation index.Conclusion. Plasma IL-8, LDH, and HBDH levels and IL-6/IL-10 ratio could be helpful for early evaluation of the severity and predicting fatal outcomes in AIDS PCP patients.

Bacterial translocation and the development of sepsis after orthotopic liver transplantation (OLT) may be promoted by immunological damage to the intestinal mucosa or by quantitative and qualitative changes in intestinal microbiota. This study monitored structural shifts of gut microbiota in rats with OLT using PCR-denaturing gradient gel electrophoresis (DGGE) and real-time quantitative PCR (RT-qPCR). RT-qPCR targets six major microorganisms (Domain Bacteria, Bacteroides, Bifidobacteria, Enterobacteriaceae, Lactobacillus and Clostridium leptum subgroup). Isograft, Allograft and Sham model were studied. Bacterial translocation to host organs and plasma endotoxin were determined. Alteration in gut microbiota was associated with the elevation of plasma endotoxin and a higher rate of bacterial translocation (BT) to liver in rats with acute rejection. Dynamic analysis of DGGE fingerprints showed that the gut microbiota structure of animals in the three groups was similar before the operation. But significant alterations in the composition of fecal microbiota in Allograft group were observed at 1 and 2 weeks after the OLT. The acute rejection was accompanied by the shifts of gut microbiota towards members of Bacteroides and Ruminococcus. Results from RT-qPCR indicated that Bacteroides significantly increased at 2 weeks after the OLT, whereas numbers of Bifidobacterium spp. decreased at 1 week and recovered at 2 weeks after the OLT. In summary, our data showed that rats with acute rejection after OLT exhibited significant structure shifts in the gut microbiota which dominant by overgrowth of Bacteroides and Ruminococcus, and these were associated with elevation of plasma endotoxin and higher rate of BT.

The present study investigated the alteration of intestinal microbiota, innate immunity-related genes, and bacterial translocation in rats with cirrhosis and liver transplantation.Specific pathogen-free Sprague-Dawley rats were randomized into 4 groups: (1) normal controls (N); (2) liver cirrhosis (LC); (3) normal control groups with liver transplantation (LTN); and (4) liver cirrhosis with liver transplantation (LTC). We examined plasma endotoxin, bacterial tacslocation, denaturing gradient gel electrophoresis (DGGE) profile of intestinal mucosa-associated bacteria, abundance of key bacterial populations, and expression of innate immunity-related gene.The LTC and LC group, showed higher endotoxin levels (1.08±0.73 EU/mL and 0.74±0.70 EU/mL, respectively) than the N group (0.27±0.13 EU/mL; P.05). the incidence of bacterial translocation (BT) to liver and mesenteric lymph nodes (MLN), and the number of total bacteria were increased significantly in the LTC and LC groups compared with the N group (P.05). The counts of Lactobacilli and Bacteroides were lower, whereas Enterobacteria were higher in the LC than the N group (P.05). Mucins (MUC2, MUC3) and Toll-like receptors (TLR2, TLR4) messenger RNA (mRNA) expression were significantly higher in the LC and LTC groups than the N group (P.05). The marked difference between the groups in the overall structure of the bacterial community was also generated by DGGE profiles.Liver cirrhosis disturbs intestinal microbiota and innate immunity-related genes, which contributes to endotoxemia and bacterial translocation. These had not completely recovered in cirrhotic rats until 1 month after orthotopic liver transplantation.

The seroprevalence of Kaposi's sarcoma-associated herpesvirus (KSHV) in Chinese patients with chronic hepatitis B, the relationship between KSHV and hepatitis B virus (HBV) infection, and the influence of glycyrrhizic acid on KSHV replication in vivo are undefined. Plasma was collected from 211 patients with chronic hepatitis B. Antibody to KSHV ORF65 was evaluated by ELISA, and real-time PCR was used to quantify KSHV DNA and HBV DNA. The KSHV ORF65 positivity rate in patients with chronic hepatitis B was found to be 28% (59/211): 27.3% (44/161) in males and 30% (15/50) in females (P0.05). The seroprevalence of KSHV increased with age until reaching the highest rate (37.1%) in the 31-40 years age group. HBV DNA loads in patients with chronic hepatitis B infected with KSHV were higher than those without KSHV infection (9.2 log (10) IU/ml vs. 7.8 log (10) IU/ml, P0.05). The average KSHV DNA loads in patients with HBV genotype B, C, and mixed (B/C) were 409.1, 484.5, and 352 copies/ml, respectively (P0.05). Patients treated with glycyrrhizic acid had lower KSHV DNA levels than those without therapy (204.7 copies/ml vs. 533.9 copies/ml, P0.05). The KSHV ORF65 positivity rates tended to increase with age, but were not related to gender or HBV genotypes. The data indicated the interaction between KSHV and HBV, and the inhibiting effect of glycyrrhizic acid on KSHV replication in patients with chronic hepatitis B.

The epidemiology and strain features of Kaposi's sarcoma-associated herpesvirus (KSHV) in transplant recipients and HIV-infected patients in China is not well described.To determine KSHV seroprevalence and characterize strains of KSHV in transplant recipients and HIV-infected patients in China.Patients were screened for KSHV infection using a KSHV open reading frame (ORF) 65 recombinant protein-dependent ELISA. The 172bp fragment of KSHV ORF26 was amplified by nested PCR and bilaterally sequenced from KSHV-seropositive peripheral blood mononuclear cells. ORF26 gene diversity and cladograms were analyzed and compared to predominant international strains.KSHV seroprevalence in renal transplant recipients [41% (44/107)] and HIV-infected patients [39% (29/74)] were significantly higher than in healthy people [13% (16/122)] (P0.01), but KSHV seroprevalence in liver transplant recipients [15% (5/33)] was similar to that of healthy people (P0.05). ORF26 DNA was detected in 47% (23/49) of KSHV-seropositive organ transplant recipients; 28% (8/29) of KSHV-seropositive HIV-infected patients; and 19% (3/16) of KSHV-seropositive healthy people. Organ transplant recipients had a significantly higher rate of KSHV DNAemia than healthy people (P0.05). The 13 KSHV strains that were analyzed were genetically close to the Hungary AY707887 strain and belonged to the A3 subtype.The seroprevalence of KSHV was elevated in renal transplant recipients and HIV-infected patients in China, and KSHV DNAemia was more common in these patients and liver transplant recipients. KSHV strains from Zhejiang Province belonged to the A3 subtype.

Immune reconstitution inflammatory syndrome (IRIS) is a severe clinical complication emerging during the initial months of antiretroviral therapy (ART) in HIV infected patients. Mycobacterium avium complex (MAC) associated IRIS typically develops in severely immunosuppressed individuals, who have an excellent response to ART. Cutaneous MAC infection is uncommon. The skin lesions almost invariably responded well to specific anti-mycobacterial treatment for MAC infection in IRIS patients. Here, we report a case whose cutaneous MAC lesions continue exacerbation despite with anti-mycobacterial treatment. But rapid clinical remission occurred after thalidomide adding to the antituberculosis treatment. Furthermore, immunomodulatory effect of thalidomide on cytokine levels such as decreasing of TNF-α, increasing of IL-4 and IL-6 were observed in the patient. Thalidomide may be an effective treatment for cutaneous MAC infection during IRIS. This is the first report of talihdomide treatment for cutaneous MAC infection during IRIS, and needs further mechanism and clinical trial investigation in this regard.

The previous studies all focus on the effect of probiotics and antibiotics on infection after liver transplantation. Here, we focus on the effect of gut microbiota alteration caused by probiotics and antibiotics on hepatic damage after allograft liver transplantation. Brown-Norway rats received saline, probiotics, or antibiotics via daily gavage for 3 weeks. Orthotopic liver transplantation (OLT) was carried out after 1 week of gavage. Alteration of the intestinal microbiota, liver function and histopathology, serum and liver cytokines, and T cells in peripheral blood and Peyer's patch were evaluated. Distinct segregation of fecal bacterial diversity was observed in the probiotic group and antibiotic group when compared with the allograft group. As for diversity of intestinal mucosal microbiota and pathology of intestine at 2 weeks after OLT, antibiotics and probiotics had a significant effect on ileum and colon. The population of Lactobacillus and Bifidobacterium in the probiotic group was significantly greater than the antibiotic group and the allograft group. The liver injury was significantly reduced in the antibiotic group and the probiotic group compared with the allograft group. The CD4/CD8 and Treg cells in Peyer's patch were decreased in the antibiotic group. The intestinal Treg cell and serum and liver TGF-β were increased markedly while CD4/CD8 ratio was significantly decreased in the probiotic group. It suggested that probiotics mediate their beneficial effects through increase of Treg cells and TGF-β and deduction of CD4/CD8 in rats with acute rejection (AR) after OLT.

The importance of diseases of tomato caused by begomoviruses is increasing worldwide. Here, we report that several begomoviruses are associated with tomato leaf curl disease in Yunnan province, China. 14 tomato samples showing leaf curl symptoms were collected in three districts in Yunnan, and they fell into four groups according to their reaction with a panel of 16 monoclonal antibodies in TAS-ELISA. Comparison of partial DNA-A sequences amplified with degenerate primers confirmed the existence of several types of begomoviruses. The complete DNA-A sequences of 4 isolates (Y25, Y41, Y72, Y161), corresponding to the four groups, were determined. Sequence comparisons and phylogenetic analysis revealed that they corresponded to four previously identified begomoviruses. Groups I, II and IV are most closely related to Tomato yellow leaf curl China virus (TYLCCNV), Tobacco curly shoot virus (TbCSV) and Tobacco leaf curl Yunnan virus (TbLCYNV), respectively, while Group III shows close relationships with Tomato yellow leaf curl Thailand virus (TYLCTHV). In addition, all isolates in Groups I and III were found to be associated with DNAbeta molecules, while satellite DNA was not found in virus isolates in Groups II and IV. The complete DNAbeta sequences of three isolates from Group III (Y72, Y77, Y79) were determined. Sequence analysis showed that Y72beta, Y77beta and Y79beta seem to be different from other characterised DNAbeta, sharing the highest nucleotide sequence identity with DNAbeta of TbCSV.

Virus isolate Y23V, obtained from squash showing leaf curl symptoms in Yunnan, China, was readily differentiated from four studied Chinese begomovirus isolates in reactions with a set of monoclonal antibodies raised against begomoviruses. The complete nucleotide sequence (2714 nts) of the DNA-A-like molecule of Y23V was determined. The DNA-A of Y23V is most closely related to that of tomato yellow leaf curl Thailand virus-[1] (TYLCTHV-[1]) (84% sequence identity). However, the AC1 and AC4 gene of Y23V DNA-A resembled to Pepper leaf curl virus from Bangladesh (PepLCBDV). The DNA-A of Y23V has three distinct regions: the region from 74-2071 nts is 95% identical to TYLCTHV-[1] excluding a 27 nt deletion; the following 386 nts are 91% identical to PepLCBDV and the rest of the DNA-A is not closely related to any reported begomovirus. Y23V, therefore, is considered to have arisen by recombination. The 84% sequence identity of Y23V with TYLCTHV-[1] allows Y23V to be considered as a distinct begomovirus species, for which the name squash leaf curl Yunnan virus (SLCYNV) is proposed.