Genetic diversity of Chinese rabies viruses: evidence for the presence of two distinct clades in China.

There have been three major rabies epidemics in China since the 1950s. To gain more insights into the molecular epidemiology of rabies viruses (RVs) for the third (the current) epidemic, we isolated RV from dogs and humans in major endemic areas, and characterized these isolates genetically by sequencing the entire glycoprotein (G) gene and the G-L non-coding region. These sequences were also compared phylogenetically with RVs isolated in China during previous epidemics and those around the world. Comparison of the entire G genes among the Chinese isolates revealed up to 21.8% divergence at the nucleotide level and 17.8% at the amino acid level. The available Chinese isolates could be divided into two distinct clades, each of which could be further divided into six lineages. Viruses in clade I include most of the Chinese viruses as well as viruses from southeast Asian countries including Indonesia, Malaysia, the Philippines, Thailand, and Vietnam. The viruses in the other clade were found infrequently in China, but are closely related to viruses distributed worldwide among terrestrial animals. Interestingly, most of the viruses isolated during the past 10 years belong to lineage A viruses within clade I whereas most of the viruses isolated before 1996 belong to other lineages within clades I and II. Our results indicated that lineages A viruses have been predominant during the past 10 years and thus are largely responsible for the third and the current epidemic in China. Our results also suggested that the Chinese RV isolates in clade I share a common recent ancestor with those circulating in southeast Asia.

Investigation of the role of healthy dogs as potential carriers of rabies virus.

To investigate whether healthy animals are potential carriers of rabies virus in China, 153 domestic dogs were collected from a rabies enzootic area, Anlong county in Guizhou Province, and monitored for 6 months. Initially, findings of rabies virus antigen in the saliva of 15 dogs by an enzyme-linked immunosorbent assay (ELISA) test suggested they might be carriers. These 15 dogs were kept under observation for 6 months. None of the dogs showed any clinical signs of rabies during the observation period. Moreover, using the ELISA test alone, detection of rabies virus antigen in saliva of some animals was not consistent during the observation period. However, none of the saliva samples collected either at the time of acquisition or during the observation period was found to be positive for rabies virus RNA by reverse transcriptase-polymerase chain reaction (RT-PCR). Furthermore, neither viral antigen nor viral RNA was detected in the brain samples collected at the time of euthanasia. These results do not provide support for the contention that healthy dogs act as carriers in rabies. Caution is urged when preliminary and nondefinitive tests, such as ELISA, are used to infer clinical status related to rabies.

Avian influenza viruses (AIVs) H9N2 are in the course of reassorting into novel AIVs.

In 2013, two episodes of influenza emerged in China and caused worldwide concern. A new H7N9 avian influenza virus (AIV) first appeared in China on February 19, 2013. By August 31, 2013, the virus had spread to ten provinces and two metropolitan cities. Of 134 patients with H7N9 influenza, 45 died. From then on, epidemics emerged sporadically in China and resulted in several victims. On November 30, 2013, a 73-year-old woman presented with an influenza-like illness. She developed multiple organ failure and died 9 d after the onset of disease. A novel reassortant AIV, H10N8, was isolated from a tracheal aspirate specimen that was obtained from the patient 7 d after onset. This case was the first human case of influenza A subtype H10N8. On 4 February, 2014, another death due to H10N8 avian influenza was reported in Jiangxi Province, China.

Spontaneous ruptured primary hepatic angiosarcoma coincident with Schistosoma Japonica liver fibrosis.

AIM: We report herein a case of spontaneous ruptured primary hepatic angiosarcoma coincident with Schistosoma Japonica liver fibrosis and review the correlative literature. METHODS: The resected specimen was examined by histopathological and immunohistochemical evaluation. RESULTS: The final diagnosis was spontaneous ruptured primary hepatic angiosarcoma coincident with Schistosoma Japonica liver fibrosis Conclusion: Considering the nature of primary hepatic angiosarcoma, in particular the ruptured hepatic angiosarcoma, it is obviously desirable to avoid any unnecessary delay or definitive surgical treatment. It is presumed that angiosarcoma in the liver has a possible association with S. japonicum and the deposition of ovae in liver.

Protective effect of curcumin against liver warm ischemia/reperfusion injury in rat model is associated with regulation of heat shock protein and antioxidant enzymes.

AIM: To investigate the hypothesis that the protective effects of curcumin in hepatic warm ischemia/reperfusion (I/R) injury are associated with increasing heat shock protein 70 (Hsp70) expression and antioxidant enzyme activity. METHODS: Sixty Sprague-Dawley male rats were randomly divided into sham, I/R, C + I/R groups. The model of reduced-size liver warm ischemia and reperfusion was used. Curcumin (50 mg/kg) was administered by injection through a branch of superior mesenteric vein at 30 min before ischemia in C + I/R group. Five rats were used to investigate the survival during 1 wk after operation in each group. Blood samples and liver tissues were obtained in the remaining animals after 3, 12, and 24 h of reperfusion to assess serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), liver tissue NO(2)(-) + NO(3)(-), malondialdehyde (MDA) content, superoxide dismutase (SOD), catalase (CAT), nitricoxide synthase (NOS) and myeloperoxidase (MPO) activity, Hsp70 expression and apoptosis ratio. RESULTS: Compared with I/R group, curcumin pretreatment group showed less ischemia/reperfusion-induced injury. CAT and SOD activity and Hsp70 expression increased significantly. A higher rate of apoptosis was observed in I/R group than in C + I/R group, and a significant increase of MDA, NO(2)(-) + NO(3)(-) and MPO level in liver tissues and serum transaminase concentration was also observed in I/R group compared to C + I/R group. Curcumin also decreased the activity of inducible NO synthase (iNOS) in liver after reperfusion, but had no effect on the level of endothelial NO synthase (eNOS) after reperfusion in liver. The 7 d survival rate was significantly higher in C + I/R group than in I/R group. CONCLUSION: Curcumin has protective effects against hepatic I/R injury. Its mechanism might be related to the overexpression of Hsp70 and antioxidant enzymes.

Molecular characterization of rabies virus isolates in China during 2004.

Human rabies cases have been on the rise during the past few years in China and a total of 2651 cases were reported in 2004. To better understand the current rabies epidemics in China, we isolated rabies viruses from dogs and humans from five provinces and characterized these isolates genetically by sequencing the entire nucleoprotein (N) gene. Comparison of the N genes among these isolates revealed 86.6-99.9% homology and these viruses can be grouped into three lineages. Phylogenetic analysis indicates that all the Chinese isolates have a close relationship with viruses circulating in Asian canine population. When compared with rabies viruses isolated previously, the three lineages were similar to three of the four groups. Thus, our data suggest that rabies viruses currently circulating in China were similar, if not identical, to those reported in the previous epidemics.

Primary malignant fibrous histiocytoma of the small bowel: a report of an additional case in duodenum.

AIMS: We report herein an additional case of primary malignant fibrous histiocytoma (MFH) in the duodenum and provide a review of the existing literature. METHODS AND RESULTS: A 61-yr-old Chinese man was admitted to our hospital with symptoms of melena, anorexia, and weight loss. An abdominal computed tomography (CT) and gastrointestinal barium meal examination demonstrated a tumor of the duodenum suggestive of primary malignancy. The tumor was successfully treated by pancreaticoduodenectomy. It was histopathologically and immunohistochemically diagnosed to be a storiform-type primary MFH of the duodenum. There have been a total of 40 cases of primary malignant fibrous histiocytoma of the small bowel documented in the literature including our Chinese cases. CONCLUSION: Primary malignant fibrous histiocytoma of the small bowel, especially in the duodenum is extremely rare. The final diagnosis is made only after pathological and immunopathological examination of the tumor. The malignant potential of such tumors is high. The prognosis may be mainly dependent on the invasion and metastasis of tumor, while tumor size is irrelevant. The treatment should be surgery if possible. Early surgical intervention may be the best form of management that may offer the patient good result.

Intraoperative radiofrequency thermal ablation combined with portal vein infusion chemotherapy and transarterial chemoembolization for unresectable HCC.

BACKGROUND/AIMS: We conducted a preliminary study to determine the feasibility of therapy consisting of intraoperative radiofrequency thermal ablation combined with portal vein infusion chemotherapy and transarterial chemoembolization (IRFAPA) for unresectable hepatocellular carcinoma (HCC). METHODOLOGY: Between September 2001 and June 2004, 34 patients with unresectable HCC were enrolled into a prospective study. 18 cases underwent IRFAPA (group I and 16 cases underwent percutaneous RF ablation (PRFA, group II). Patients' outcomes for IRFAPA and PRFA were recorded and compared. RESULTS: Patients undergoing IRFAPA or PRFA were similar in age, liver function, tumor size, serum AFP, distribution of tumor, mortality, complication and complete ablation rates. In five patients in group II seven new lesions were found during operation. The rate of distant intrahepatic recurrence between the two groups had differences (11.1% vs. 50.0%, P=0.023) although the cumulative recurrence-free survival between the two groups had no differences (P=0.7808). There was a significant difference in the overall survival (P=0.0407). The 1-year and 3-year cumulative overall survival rate was 87.5% and 73.3%, 52.2% and 20.4% in group I and group II, respectively. CONCLUSIONS: IRFAPA is an effective and safe procedure for unresectable HCC. IRFAPA is preferred to PRFA therapy if the patients' conditions can tolerate laparotomy.

Comparison of data-fitting models for schistosomiasis: a case study in Xingzi, China.

When modelling prevalence data, epidemiological studies usually employ either Gaussian, binomial or Poisson models. However, reasons are seldom given in the literature why the chosen model was felt to be the most appropriate. In this study, we compared all three models for fitting schistosomiasis risk in Xingzi county, Jiangxi province, People's Republic of China. Parasitological data from conventional surveys were available for 36,208 individuals aged between 6 and 65 years from 42 sampled villages and used in combination with environmental data to map the spatial patterns of schistosomiasis risk. The results show that the Poisson model fitted the data best and this model identified the role of environmental risk factors in explaining the geographical variation of schistosomiasis risk. These factors were further used to develop a predictive map, which has important implications for the control and eventual elimination of schistosomiasis in the People's Republic of China.

Analysis of HER2 gene amplification and protein expression in esophageal squamous cell carcinoma.

The HER2 gene, which is located on chromosomes 17, is a therapeutic target for cancer. Amplification of HER2 has been described in several tumor types. However, few studies of HER2 gene amplification and protein expression in esophageal carcinoma have been conducted. This study was to investigate the relationship between the expression of HER2/neu and the clinical characteristics, including survival rate, of esophageal squamous carcinoma. The clinical data of 145 patients admitted in Renmin Hospital of Wuhan University, from 2000 to 2005, were reviewed. The HER2 protein expression and gene status in 145 esophageal carcinomas were evaluated using immunohistochemistry and fluorescence in situ hybridization. The survival rate was calculated by the Kaplan-Meier method and the log-rank test using SPSS13.0 software. Compared to normal esophageal epithelium (23/95, 24.2%), HER2 protein was overexpressed in most esophageal squamous carcinoma tissues (60/145, 41.4%), of which 45 (31.0%) were 2+ and 15 (10.4%) were 3+, HER2 overexpression associated significantly with HER2 gene amplification. There is a correlation between the overexpression of HER2 and the differentiation of the carcinoma, the HER2 gene amplification and the differentiation of the carcinoma and the tumor stage. According to univariate analysis, there was a significant difference in survival rates when cases with and without HER-2/neu overexpression or amplification were compared. HER-2/neu amplification/overexpression may be used as an independent prognostic factor in patients with esophageal squamous cancer, and patients with HER-2/neu amplification/overexpression might be potential candidates for new adjuvant therapies that involve the use of humanized monoclonal antibodies.

Microvessel damage may play an important role in tumoricidal effect for murine h(22) hepatoma cells with hyperthermia in vivo.

BACKGROUND: In the present study, murine H(22) hepatoma cells were provided hyperthermia with different thermal dose in vitro and in vivo, thereafter we investigated the apoptosis, necrosis rates, and intratumoral microvessel density (MVD) to determine that microvessel damage plays an important role in the tumoricidal effect of hyperthermia. METHODS: H(22) hepatoma cells were inoculated in the right hind legs of mice with immunosuppression. Local hyperthermia was administered to these mice for 15, 30, and 45 min, respectively. After hyperthermia, some mice with heat treatment of 30 min were killed at 3, 6, 12, 24, 48, 72, and 96 h after operation and others were immediately sacrificed. All tumor tissues were removed. They were analyzed for the death rate of tumor cells by flow cytometer (FCM) and observed MVD by immunohistochemistry. H(22) hepatoma cells in vitro were also given hyperthermia for 15, 30, and 45 min, respectively, and analyzed for the death rate by FCM. RESULTS: Most of the dead cells were apoptotic cells in the initiation phase of hyperthermia, then the necrosis rates rose gradually. The difference of death rates between in vivo and in vitro was significant for hyperthermia for 15 min, 30 min, and 45 min (P < 0.05). A strong positive linear correlation (r = -0.879) was observed between the death rate of tumor cells and MVD. CONCLUSION: Our study has shown that microvessel damage may play an important role in tumoricidal effect of hyperthermia.

[An investigation on hepatitis E virus infection status among livestock in Xi'an area].

OBJECTIVE: To explore the carrier state of hepatitis E virus(HEV) in livestock in Xi'an area. METHODS: Bile samples from swine, canine, sheep and cow were collected from a local slaughtering house. Reverse transcriptase nested polymerase chain reaction (RT-nPCR) was employed to amplify the ORF2 region in HEV RNA genome. All positive samples were sequenced and compared with data from GenBank. Homology analysis was conducted based on the outcome of sequencing. RESULTS: 194, 178, 79 and 191 bile samples from swine, canine, cow and sheep were collected. Positive rates with RT-nPCR method in these domestic animals were 4.10%, 0%, 0% and 0% respectively. Genetic distance analysis indicated that strains being identified were close to genotype IV of HEV, then genotype I, II and III in nucleic acid. Same outcome was shown by the same analysis on amino acid. CONCLUSION: Swine was the only reservoir of HEV in livestock and genotype IV was the prevalent genotype.

Primary epithelioid angiosarcoma of the male breast: report of a case.

We report a case of primary epithelioid angiosarcoma of the male breast. The patient was a 20-year-old Chinese man who presented with a huge tumor just below the left nipple. Histopathological examination and immunohistochemical analysis confirmed a diagnosis of primary epithelioid angiosarcoma of the male breast, without axillary lymph node metastasis. We review the relevant literature on this rare malignant tumor.

The protective effects of 17beta-estradiol on hepatic ischemia-reperfusion injury in rat model, associated with regulation of heat-shock protein expression.

BACKGROUND: Ischemia-reperfusion (I/R) injury, which was commonly seen in the field of hepatic surgical intervention, impaired liver regeneration and predisposed to liver failure. Previous studies have shown gender dimorphic response of the liver for various hepatic stresses including I/R injury, hemorrhagic shock-resuscitation, liver cirrhosis, endotoxemia, and chronic alcoholic consumption, and demonstrated gender dimorphism in hepatocellular dysfunction after experimental trauma and hemorrhage. The objective of this study was to examine the hypothesis that the protective effects of 17beta-estradiol (E2) in hepatic I/R injury were associated with increasing heat-shock protein 70 expression. MATERIALS AND METHODS: Sprague-Dawley male and female rats were randomly divided into male and female sham, I/R, and E2 + I/R groups. The model of reduced-size liver ischemia and reperfusion was used. Except for the sham-operated groups, all rats were subjected to 70% liver ischemia for 45 min followed by resection of the remaining 30% nonischemic lobes and reperfusion of ischemic tissue. For each group, five rats were used to investigate the survival during a week after operation; blood samples and liver tissues were obtained in the remaining animals after 3, 12, and 24 h of reperfusion to assess serum alanine aminotransferase, aspartate aminotransferase, liver tissue NO(2)(-) + NO(3)(-), malondialdehyde content, superoxide dismutase, nitric oxide synthase, and myeloperoxidase activity, Hsp70 expression, and apoptosis ratio. RESULTS: Compared with I/R groups, male and female E2 + I/R groups showed less I/R-induced injury, and SOD and eNOS activity and Hsp70 expression were increased significantly (P < 0.01). A higher rate of apoptosis was observed in the I/R group versus the E2 + I/R group, a significant increase of MDA, NO(2)(-) + NO(3)(-), and MPO of liver tissues and serum transaminase were also observed in the I/R group versus the E2 + I/R group. The survival rate was significantly higher in the male E2 + I/R group than in the male I/R group. CONCLUSION: E2 pretreatment had protective effects on liver in hepatic I/R injury. The mechanism of this protection might be related to overexpression of Hsp70.

[Molecule character of rabies virus in 3 counties in Hunan China].

OBJECTIVE: To study the rabies molecular biology features in animals between high incidence area of rabies and no rabies cases area in Hunan. METHODS: detect saliva of dogs and brains of dogs and cats by direct immunofluorescence assay, review positive samples by RT-PCR, sequencing extract RNA virus for genetic analysis. RESULTS: 12 were detected rabies virus antigen and positive nucleoside acid in 82 dogs from Wugang city also 1 in 17 from Dongkou county; the positive rate: Wugang 14.63 percent, Dongkou 5.88 percent. No rabies virus was detected in 67 samples of dog brains from Fenghuang County. Also none in 28 samples of cat brains. Amplificating N gene of rabies virus from positive samples of dog brain's tissue (No Wg13, Dk13) by RT-PCR, it shows that homology of nucleoside acid between two strain of virus is 99.4 percent; also 99.1 percent of amino acid. The homology of nucleoside acid (amonio acid) among Wg13 stain and Chinese strain CTN and aG strain is 89.4 percent (98.2 percent) and 86.1 percent (95.1 percent); The homology of nucleoside acid (amonio acid) among Dk13 stain Chinese strain CTN and aG strain is 89.1 percent (98.0 percent), 86.1 percent (94.9 percent).Compare with isolated rabies virus from abroad, the homology between two strains and Indonesia is 92.8 percent and 93.2 percent, the most similar of them. The strains isolated from other countries including Japan, Sri Lanka and India are relatively lower; The sequence of gene Wg13 and Dk13 were taken replacement of amino acid. CONCLUSION: Two strains are belong to type I rabies virus, comparing its N gene with current using vaccine strains, both are in same group, and homology are relatively higher.

[Analyses on the isolation and characterization of canine rabies virus isolates in Henan].

OBJECTIVE: To analyze the genetic characteristics of two rabies virus isolates from Henan province and to compare their relations hip with known rabies virus isolates and vaccine strains. METHODS: Rabies viral antigens were detected in 100 canine brains by immunofluorescence assay method. Rabies virus was isolated through inoculating the suspensions of positive brains into suckling mice. N gene and G were amplified by RT-PCR and sequenced. Phylogenetic trees were constructed for the analysis on genetic characteristics of rabies virus. RESULTS: Two rabies virus strains were isolated (Henan Hb1 and Henan Sq1). Data from sequential comparison revealed that the nucleotide and amino acid identities of N and G gene between the two isolates were 99.3% and 98.9%, and 98.7% and 98.4% respectively. The two isolates were more closely related to CTN, with the homogeny of N gene and G gene as 89.1% and 85.6%-85.7% at the nucleotide level, but 97.6%-98.0% and 92.3% at the amino acid level respectively, than to other vaccine strains. When comparing with other known viruses including Chinese isolates, the two stains shared closer identity with the isolates from Indonesia, and the rates of homogeny of N and G gene were 92.1%-93.2% and 91.9%-92.1% at the nucleotide level, 97.5%-98.6% and 96.0%-96.2% at the amino acid level, respectively. Data from the deduced amino acid sequences revealed that some amino acid residues including the residues in the N and G antigenic sites were substituted in the two isolates. Furthermore, phylogenetic analysis showed that the two isolates were also more closely related to the strains from Indonesia and vaccine strain CTN than to any other known street viruses and vaccine strains. CONCLUSION: Both Henan Hbl and Henan Sql belonged to genotype 1. However, the N and G gene diverged from known street viruses and vaccine strains at either nucleotide level or amino acid level.

[The survey of 21 cases of human rabies in Anlong county in Guizhou province].

OBJECTIVE: To analyze 21 cases of rabies from February 8 to May 1 in 2004 in Anlong county in Guizhou province, and to explore the possible factors causing the epidemics. METHODS: 21 cases of rabies were investigated. The canine brains were collected, and indirect immunofluorescence assay was used to detect rabies virus antigen in the brains. RESULTS: From February 8 to May 1st in this year, 21 cases of rabies were reported, with 5.12/100,000 of the prevalence rate. Dogs accounted for 20 cases, and cat for one case. Median incubation period was 36.52 days, but less than 15 days in 6 cases. Among 21 cases, 17 cases were not correctly treated, and 9 cases received vaccine, with 3 cases vaccinated in time. No cases received passive immunization of antirabies serum or human antirabies immune globulin. A total of 73 dogs' brains were examined for rabies viral antigen by IFA, and 9 (12.33%) were positive. CONCLUSION: The increasing number of dog, high rate of virus carrier in dogs, incorrect treatment of the wound, and as well as low inoculating rate of rabies vaccine might be responsible for the outbreak of rabies in Anlong county.