Evolving Therapeutic Landscape of Intracerebral Hemorrhage: Emerging Cutting-Edge Advancements in Surgical Robots, Regenerative Medicine, and Neurorehabilitation Techniques.

Intracerebral hemorrhage (ICH) is the most serious form of stroke and has limited available therapeutic options. As knowledge on ICH rapidly develops, cutting-edge techniques in the fields of surgical robots, regenerative medicine, and neurorehabilitation may revolutionize ICH treatment. However, these new advances still must be translated into clinical practice. In this review, we examined several emerging therapeutic strategies and their major challenges in managing ICH, with a particular focus on innovative therapies involving robot-assisted minimally invasive surgery, stem cell transplantation, in situ neuronal reprogramming, and brain-computer interfaces. Despite the limited expansion of the drug armamentarium for ICH over the past few decades, the judicious selection of more efficacious therapeutic modalities and the exploration of multimodal combination therapies represent opportunities to improve patient prognoses after ICH.

Environmental surveillance reveals co-circulation of distinctive lineages of enteroviruses in southwest China's border cities, 2020-2022.

AIMS: Enteroviruses are significant human pathogens associated with a range of mild to severe diseases. This study aims to understand the diversity and genetic characterization of enteroviruses circulated in southwest China's border cities by using environmental surveillance. METHODS AND RESULTS: A total of 96 sewage samples were collected in three border cities and a port located in Yunnan Province, China from July 2020 to June 2022. After cell culture and VP1 sequencing, a total of 590 enterovirus isolates were identified, belonging to 21 types. All PV strains were Sabin-like with ≤6 nucleotide mutations in the VP1 coding region. Echovirus 6, echovirus 21 (a rare serotype in previous studies), and coxsackievirus B5 were the predominant serotypes, which accounted for 21.19%, 18.31%, and 13.39% of the total isolates, respectively. The prevalence of the common serotypes varied across different border cities and periods. Phylogenetic analysis revealed the presence of multiple evolutionary lineages for E21, E6, and E30, some of which formed distinct branches. CONCLUSIONS: High diversity of enteroviruses and distinct lineages of predominant serotypes circulated in southwest China's border cities.

α-Linolenic acid-regulated testosterone biosynthesis via activation of the JNK-SF-1 signaling pathway in primary rooster Leydig cells.

As a functional fatty acid, α-linolenic acid (ALA) is essential in promoting animal testosterone biosynthesis. This study investigated the effects of ALA on testosterone biosynthesis and the possible mechanism underlying the signaling pathway in primary Leydig cells of the rooster. METHODS: Primary rooster Leydig cells were treated with ALA (0, 20, 40, or 80 µmol/L) or pretreated with a p38 inhibitor (50 µmol/L), a c-Jun NH2-terminal kinase (JNK) inhibitor (20 µmol/L), or an extracellular signal-regulated kinase (ERK) inhibitor (20 µmol/L) before ALA treatment. Testosterone content in the conditioned culture medium was detected using an enzyme-linked immunosorbent assay (ELISA). The expression of steroidogenic enzymes and JNK-SF-1 signaling pathway factors was detected using real-time fluorescence quantitative PCR (qRT-PCR). RESULTS: Supplementation with ALA significantly increased testosterone secretion within culture media (P < 0.05), and the optimized dose was 40 µmol/L. Compared with the control group, steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (P450scc), and 3ß-hydroxysteroid dehydrogenase (3ß-HSD) mRNA expression significantly increased (P < 0.05) in the 40 µmol/L ALA group; 17-hydroxylase/c17-20 lyase (P450c17) and p38 mRNA expressions were not significantly different in the 40 µmol/L ALA group; ERK and JNK mRNA expressions were significantly upregulated (P < 0.05) in 40 µmol/L ALA group. In the inhibitor group, testosterone levels were significantly downregulated (P < 0.05). Compared with the 40 µmol/L ALA group, StAR, P450scc, and P450c17 mRNA expressions were significantly decreased (P < 0.05), and 3ß-HSD mRNA expression in the p38 inhibitor group did not change; StAR, P450scc, and 3ß-HSD mRNA expressions were significantly decreased (P < 0.05), and P450c17 mRNA expression in ERK inhibitor group did not change; StAR, P450scc, 3ß-HSD, and P450c17 mRNA expressions were significantly decreased (P < 0.05) in JNK inhibitor group. Additionally, the increased steroidogenic factor 1 (SF-1) gene expression levels induced by ALA were reversed when the cells were pre-incubated with JNK and ERK inhibitors. The levels in the JNK inhibitor group were significantly lower than those in the control group (P < 0.05). CONCLUSION: ALA may promote testosterone biosynthesis by activating the JNK-SF-1 signaling pathway to upregulate StAR, P450scc, 3ß-HSD, and P450c17 expression in primary rooster Leydig cells.

RhoA improves cryopreservation of rooster sperm through the Rho/RhoA-associated kinase/cofilin pathway.

Cryopreservation of rooster sperm leads to relatively low semen quality due to cytoskeletal damage during the freeze-thawing process. This study aimed to explore how the addition of RhoA recombinant protein affected the viability and subcellular structure of rooster sperm after freeze-thawing and elucidated the molecular mechanisms of sperm cryopreservation. Semen quality and acrosome integrity testing revealed that the addition of 0.5 µg/mL RhoA recombinant protein to the cryoprotectant fluid significantly increased sperm motility, survival rate, linearity, straight-line velocity, and acrosome integrity after freeze-thawing (P < 0.05). Ultrastructure analysis of cryopreserved sperm showed structural damage to the sperm plasma membrane, nuclear membrane, and tail. However, compared to the control, these structural changes were reduced upon the addition of RhoA recombinant protein to the cryoprotective fluid (P < 0.05). Western blotting revealed that the expression of Rho/RhoA-associated kinase and p-cofilin was increased, and cofilin expression was decreased after sperm cryopreservation with recombinant RhoA protein. Treatment with Y-27632, a ROCK antagonist, suppressed ROCK and p-cofilin expression and decreased semen quality, acrosome integrity, and ultrastructure integrity. In summary, we have demonstrated a cryoprotective effect in spermatozoa involving the Rho/ROCK pathway during freeze-thawing. Furthermore, the addition of 0.5 µg/mL RhoA recombinant protein to the cryoprotective fluid improved rooster semen quality and subcellular structural homeostasis after freeze-thawing via the Rho/ROCK pathway. This pathway may regulate the dynamic reorganization of the actin cytoskeleton by regulating the cofilin phosphorylation.

Molecular epidemiology of hepatitis B virus genotypes and subgenotypes in ethnic minority populations, Yunnan province, China.

The aim of our study was to determine the distribution of hepatitis B virus (HBV) genotypes and subgenotypes in ethnic minorities in Yunnan province to provide evidence supporting the theoretical basis for hepatitis B prevention and control. We obtained serum samples and demographic data from 765 individuals reported by Yunnan province who had either acute or chronic HBV infection and were from one of 20 ethnic minority populations: Achang, Bai, Brown, Tibetan, Dai, Deang, Dulong, Hani, Hui, Jingpo, Lahu, Yi, Lisu Miao, Naxi, Nu, Pumi, Wa, Yao, or Zhuang people. We sequenced the HBV DNA and determined the genotypes and subgenotypes of the isolated HBVs. We mapped the genotype and subgenotype distribution by ethnic minority population and conducted descriptive analyses. There were four genotypes among the 20 ethnic groups: genotype B (21.3% of samples), C (76.6%), D (1.8%) and I (0.3%). The most common subgenotype was C1. There were no genotype differences by gender (P = 0.954) or age (P = 0.274), but there were differences by region (P < 0.001). There were differences in genotype distribution (P < 0.001) and subgenotype distribution (P = 0.011) by ethnic group. Genotype D was most prominent in Tibet and most HBV isolates were C/D recombinant viruses. The only two genotype I virus isolates were in Zhuang people. Susceptibility and geographic patterns may influence HBV prevalence in different ethnic populations, but additional research is needed for such a determination.

Health and economic costs of an import-initiated measles outbreak in an international border area of Yunnan Province.

BACKGROUND: Measles outbreaks often require labor- and resource-intense response. A border-area measles outbreak occurred in Yunnan province that required outbreak response immunization for its containment. We report results of our investigation into the outbreak and the health sector costs of the response activities, with the goal of providing evidence for policy makers when considering the full value of vaccines and of measles elimination. METHODS: We conducted case investigations to determine sources of infection and routes of transmission. Costs of outbreak response activities incurred by health sector were determined through retrospective interviews and record reviews of staff. RESULTS: In total, 67 measles cases were confirmed, including 4 cases imported from Myanmar. Among the 33 cases aged between 8 months and 14 y old, 22 (66∙7%) had received 2 doses of MCV; 2 (6∙0%) had received 1 dose of MCV; 9 (27∙3%) had not received MCV. The first 4 cases had been infected in Myanmar, and we identified 8 transmission clusters with a total of 62 cases. Transmission among Yunnan province residents occurred in schools, family settings, and at gatherings. The overall cost to control the outbreak was $214,774, for a per-case cost of $3,206. The outbreak response vaccination campaign accounted for 64% of the total outbreak costs. CONCLUSIONS: Despite high population immunity among Yunnan province children and adolescents, an import-related measles outbreak occurred among individuals who were not vaccinated or had vaccine failure in the across-border area. The economic cost of the outbreak was substantial. Investment in a sensitive measles surveillance system to detect outbreaks in a timely manner, maintaining high population immunity to measles, and reinforcing cross-border collaboration with neighboring countries support achieving and sustaining measles elimination in the border areas of China.

Measles vaccination among children in border areas of Yunnan Province, Southwest China.

BACKGROUND: Border areas are at high risk of measles epidemics. This study aimed to evaluate the effectiveness of the implementation of the routine two-dose measles containing vaccine (MCV) program in border counties of Southwest China. METHODS: Data used in the study were derived from a cross-sectional survey among 1,467 children aged 8 to 84 months from five border counties of Yunnan Province, Southwest China in 2016. The participants were recruited using a multistage sampling method. Primary guardians of the children were interviewed to collect information on vaccination history, socio-economic status, and knowledge about immunization. Both coverage and timely coverage for the first (MCV1) and the second (MCV2) dose of MCV were calculated. The Kaplan-Meier method was performed to estimate the cumulative coverage of MCV, and Log-rank tests were adopted to compare the differences across counties and birth cohorts. Univariate and multivariate logistic regression models were used to investigate the predictors of delayed MCV1 vaccination. RESULTS: The coverage for MCV1 and MCV2 were 97.5% and 93.4%, respectively. However, only 63.8% and 84.0% of the children received MCV1 or MCV2 on time. Significant differences in the cumulative coverage were detected across counties and birth cohorts. Results of the multivariate logistic regression analysis indicated that children whose primary guardian knew the schedule of MCV were less likely to receive MCV1 late (OR = 0.63, P<0.01). For the guardians, doctors at vaccination units were the primary and also the most desired source of vaccination information. CONCLUSIONS: Although the coverage for MCV is high in border areas of Southwest China, the timeliness of MCV vaccination seems suboptimal. Tailored information from local health professionals may help to reduce untimely vaccination.

Measles immunity in the China-Myanmar border region, Lincang city, Yunnan province, 2017.

Background: A recent measles importation-related outbreak in the China-Myanmar border region required outbreak response immunization to stop the outbreak, raising the question of measles immunity in the area. We conducted two measles serological surveys to develop a seroepidemiological profile of native Chinese citizens and registered Myanmar immigrants to identify strategies to promote measles elimination in the border region.Methods: We conducted a clinic-based measles serosurvey of Myanmar registered immigrants (n = 300) and a population-based serosurvey of Chinese Lincang residents (n = 355). Serum samples were tested with enzyme-linked immunosorbent assays to assess immunity to measles.Results: We found that Myanmar immigrants had a seropositivity rate of 85.3% (95% CI: 81.3-89.4) with a GMT of 924.9 mIU/ml, and Lincang Chinese nationals had a seropositivity rate of 94.6% (95% CI:92.3-97.0) with a GMT of 1363.3 mIU/ml. Myanmar children 2-6 years of age and 7-14 years of age were more likely to be susceptible to measles than same-aged Chinese children: odds ratios 23.00 (95% CI: 5.10-103.69, P < .001) and 7.95 (95% CI: 1.66-38.01,P = .009), respectively.Conclusions: We recommend conducting a catch-up vaccination program for Myanmar children ≤15 years old.

Reduction patterns of Japanese encephalitis incidence following vaccine introduction into long-term expanded program on immunization in Yunnan Province, China.

BACKGROUND: Japanese encephalitis (JE) is a leading cause of childhood viral encephalitis both at global level and in China. Vaccination is recommended as a key strategy to control JE. In China most JE cases have been reported in southwest provinces, which include Yunnan. In this study, we quantify the epidemiological shift of JE in Yunnan Province from 2005 to 2017, covering before and after the introduction of JE vaccination into routine Expanded Program on Immunization (EPI) in 2007. METHODS: We used routinely collected data in the case-based JE surveillance system from 2005 through 2017 in Yunnan. Cases were reported from hospital and county-level Centers for Disease Control in line with the National JE Surveillance Guideline. Epidemiological data were extracted, analysed and presented in appropriate ways. Immunization coverage was estimated from actual JE doses administered and new births for each year. RESULTS: A total 4780 JE cases (3077 laboratory-confirmed, 1266 clinical and 437 suspected) were reported in the study period. Incidence of JE (per 100 000 population) increased from 0.95 in 2005 to 1.69 in 2007. With increase in vaccination coverage, incidence rates decreased steadily from 1.16 in 2009 to 0.17 in 2017. However, seasonality remained similar across the years, peaking in June-September. Banna (bordering Myanmar and Laos), Dehong (bordering Myanmar), and Zhaotong (an inland prefecture) had the highest incidence rates of 2.3, 1.9, and 1.6, respectively. 97% of all cases were among local residents. As vaccination coverage increased (and incidence decreased), proportion of JE cases among children < 10 years old decreased from 70% in 2005 to 32% in 2017, while that among adults ≥20 years old increased from 12 to 48%. There were a large number of JE cases with unknown treatment outcomes, especially in the earlier years of the surveillance system. CONCLUSIONS: The 13-year JE surveillance data in Yunnan Province showed dramatic decrease of total incidence and a shift from children to adults. Improving vaccination coverage, including access to adults at risk, and strengthening the JE surveillance system is needed to further control or eliminate JE in the province.

Measles outbreak prevention and control among adults: Lessons from an importation outbreak in Yunnan province, China, 2015.

BACKGROUND: The incidence of measles in Yunnan Province among vaccine target-age children has decreased markedly after attaining and sustaining high 2-dose coverage of measles containing vaccine (MCV) through routine immunization services and supplementary immunization activities (SIAs). Most cases of measles now occur among adults. In 2015, we investigated a measles outbreak among adults to determine transmission patterns and the potential role of nosocomial transmission. METHODS: We enhanced measles surveillance using retrospective active case search. We conducted case investigations to determine sources of infection and routes of transmission; laboratory testing included serologic and molecular diagnostic methods. RESULTS: Twenty-two outbreak-associated cases of measles were identified; most (86.36%) were among individuals 20 to 39 years of age (range, 7 months to 43 years). We interviewed 19 individuals who had acquired measles. The first 3 cases were infected in Tibet; 12 (63%) were hospital-acquired infections; 2 (11%) were community-acquired; and 2 (11%) were family-contact cases. We conducted outbreak response immunization (ORI) that provided MCV without regard to vaccination status; 10,596 residents between 20 and 40 years of age were vaccinated. A serological survey conducted during the ORI showed that 84% of the 20-40 year-olds were immune to measles prior to vaccination. Post-vaccination serological testing showed 100% were immune. CONCLUSIONS: Despite high population immunity among children and adolescents, imported measles virus transmission occurred among adults in a provincial cross-border area. Nosocomial transmission and measles immunity gaps among adults poses a threat to measles elimination and highlights the strategy of targeting adults during ORI to outbreaks with adult-to-adult transmission.

[Complete Genome Characterization of Vaccine-derived Polioviruses Isolated in Yunnan,China].

To explore the genomic characterization of 4vaccine-derived poliovirus(VDPV)strains isolated from 2acute flaccid paralysis(AFP)cases in Yunnan Province in 2010 and 2012,respectively,the complete genome sequences of the 4strains were determined. Sequence analysis revealed that the complete genome length of the type Ⅱ and type Ⅰ VDPV was 7439nt and 7441 nt, respectively. Nucleotide and amino acid sequence similarities of type II VDPV were 95.4% and 97.7%,respectively,and type I VDPV were93.9% and 97.9%,respectively as compared with those of Sabin strains. Nucleotide substitutions were found at two important attenuation sites (nt 481 and nt in type Ⅱ VDPV, and three important attenuation sites(nt480,nt2795 and nt6203)in type I VDPV. Type 2 and type 1VDPV strains had 1.0% and2.3% divergence with Sabin strains, respectively. Similarity plot analysis showed multiple recombination events in the genome of the 4strains,which showed that the recombination was common and complex. Analysis of the characteristics of VDPVs on molecular level could provide valuable information on evolutionary dynamics and lay foundation for developing scientific and feasible strategy to control VDPV.

Molecular epidemiology and recombination of human enteroviruses from AFP surveillance in Yunnan, China from 2006 to 2010.

The study represents the genetic overview of non-polio enteroviruses (NPEV) isolated from acute flaccid paralysis (AFP) cases in Yunnan Province from 2006 to 2010. Molecular typing based on VP1 nucleotide sequence was carried out on 98 NPEV isolates, and 33 serotypes were identified. EV-B was detected most frequently with an overall prevalence of 71.4%, followed by EV-A (18.4%) and EV-C (10.2%). No EV-D was identified. NPEV positive rate was higher in children <3 years of age and in summer and autumn months. Clinically, 68.4% patients presented with fever, and 16 cases (16.3%) were classified as Guillain-Barré syndrome, followed by myositis (13.3%). The phylogenetic analysis on the VP1 and 3D regions of prevalent serotypes provided evidence for recombination events among them. EV-A71, an important pathogen previously demonstrated to be associated with paralysis, had also been detected (n = 8) in this study and they all belonged to genotype C4. Great genetic divergence between Yunnan isolates and strains from other regions of the world was revealed. The findings of the study are of great importance for further research on molecular evolution of EV under the circumstance of no specialized EV surveillance system in China.

Risk factors associated with immunoprophylaxis failure against mother to child transmission of hepatitis B virus and hepatitis B vaccination status in Yunnan province, China.

OBJECTIVE: To explore the risk factors associated with immunoprophylaxis failure against mother to child transmission of hepatitis B virus (HBV) and hepatitis B vaccination status in Yunnan province, China. METHODS: Multicenter cluster sampling was used to select pregnant women who were positive for hepatitis B surface antigen (HBsAg). HBV immunoprophylaxis was carried out for the newborns. Blood samples were collected and tested for HBV markers from 7 to 10 month old infants. The factors were analyzed by univariate and logistic regression. RESULTS: A total of 2765 mothers and their infants were enrolled. The failure rate of prevention of mother to child transmission (PMTCT) was 4.12%. The rate of timely HepB1 vaccination within 24h was 98.04%, the rate of three-dose vaccination was 92.30% and the rate of hepatitis B immune globulin (HBIG) administration was 68.97%. Place of residence, maternal education, gestational age and birth weight were related to administration of HBV immunoprophylaxis. It was remarkable that the rate of HBIG administration of infants was only 63.89% with whose mothers were both HBsAg and hepatitis B e antigen (HBeAg)-positive. Further analysis showed that there were three risk factors associated with HBV immunoprophylaxis failure: mothers who were positive for HBsAg and HBeAg, maternal HBVDNA level, and HBIG administration or not. CONCLUSIONS: PMTCT of HBV was well implemented in Yunnan. However, in order to achieve optimal prevention of vertical HBV transmission, it is mandatory to make additional efforts to improve the implementation of regulatory HBV immunoprophylaxis, especially for HBsAg-positive pregnant women.

[Evaluation on the efficacy of prevention programs and relevant factors targeting mother-to-infant transmission on hepatitis B virus in Yunnan province].

OBJECTIVE: To explore the efficacy of prevention programs and relevant factors targeting mother-to-infant transmission of HBV in Yunnan province. METHODS: In Yunnan province, we selected HBsAg positive pregnant women that delivered in hospital from January 1st through June 30th, 2011. Newborns of these pregnant women were under PMTCT (prevention of mother to child treatment) program and followed. Every infant was drawn 2 ml venous blood and questionnaire survey was carried out when the baby was 7-12 month-old and completed the vaccination processes. Serum samples of them were then collected and detected on the 5 serological indicators of HBV. RESULTS: were analyzed statistically. RESULTS: There were 2 765 infants in the study program. The success rate of PMTCT was 95.88% . Rates of coverage on both timely-birth dose and 3 doses of HepB were 97.03% and 92.30% respectively. The overall vaccinated rate and timely-birth vaccinated rate on hepatitis B immunoglobulin (HBIG) were 68.97% and 94.49% respectively. The success rate of PMTCT was 97.16% after administration of passive-active immune-prophylaxis (HepB and HBIG), compared to the rate as 93.01% when vaccinated with HepB only. Significant differences were seen in the successful rates of PMTCT between combined and non-combined immunization. Either the combined or non-combined immunization, there were significant differences seen in the success rates of PMTCT regardless the positivity status of HBsAg or HBeAg, among the infected mothers. CONCLUSION: The efficacy of passive-active immune-prophylaxis program seemed to be better than the one without combined immunization. It was vitally important for the infants whose mothers' HBsAg and HBeAg status were positive, to receive regular and timely combined immunization. In order to promote the PMTCT in Yunnan province, vaccinated rate on HBIG should be further improved.

[Molecular epidemiological analysis of ECHO7 virus isolated from sewage water in Yunnan Province, China].

To investigate the epidemic and evolutionary trends of enterovirus (EV) in the external environment of Yunnan Province, China, molecular typing was performed on 4 EV strains that were isolated from environmental sewage in Yunnan. The VP1 region of isolates was amplified by RT-PCR using universal enterovirus primers, and the amplified VP1 region was sequenced for GenBank BLAST search and genotype analysis. The 4 EV strains were identified as ECHO7. Their nucleotide and amino acid homologies with the VP1 sequences of 68 ECHO7 strains retrieved from GenBank were measured by Mega software analysis. Our findings showed that ECHO7 strains from environmental sewage and population samples were in different evolutionary branches. These strains showed typical geographical and temporal differences; In addition, there were different transmission chains at the same time and in the same area. ECHO7 strains isolated from sewage water and patients with acute flaccid paralysis during the same period in Yunnan belonged to different clusters and evolved at different speeds. Special concerns are needed for this problem. Continuous molecular biological surveillance of human EV in the external environment of Yunnan will provide strong support for early warning of EV diseases.

Complete genome characterization of a novel enterovirus type EV-B106 isolated in China, 2012.

Human enterovirus B106 (EV-B106) is a recently identified member of enterovirus species B. In this study, we report the complete genomic characterization of an EV-B106 strain (148/YN/CHN/12) isolated from an acute flaccid paralysis patient in Yunnan Province, China. The new strain had 79.2-81.3% nucleotide and 89.1-94.8% amino acid similarity in the VP1 region with the other two EV-B106 strains from Bolivia and Pakistan. When compared with other EV serotypes, it had the highest (73.3%) VP1 nucleotide similarity with the EV-B77 prototype strain CF496-99. However, when aligned with all EV-B106 and EV-B77 sequences available from the GenBank database, two major frame shifts were observed in the VP1 coding region, which resulted in substantial (20.5%) VP1 amino acid divergence between the two serotypes. Phylogenetic analysis and similarity plot analysis revealed multiple recombination events in the genome of this strain. This is the first report of the complete genome of EV-B106.

Prevalence of asymptomatic Bordetella pertussis and Bordetella parapertussis infections among school children in China as determined by pooled real-time PCR: a cross-sectional study.

BACKGROUND: Studies have documented that older children and adolescents act as a reservoir of Bordetella pertussis infection for young infants who have not yet completed their primary immunization schedule. Asymptomatic pertussis infection has been reported during outbreaks. This cross-sectional study aimed to investigate whether B. pertussis and Bordetella parapertussis can colonize the nasopharynx of healthy school children, using culture and pooled real-time PCR with targets for insertion sequences IS481 and IS1001. METHODS: Nasopharyngeal (NP) swabs were taken from 629 asymptomatic school children aged 7 to 15 y in 4 counties of China during the period July-September 2011. The number of subjects included in each county ranged from 153 to 165. The 4 counties selected are located in the north, south, east, and southwest regions of China. NP swabs were inoculated onto Regan-Lowe agar for isolation of suspected Bordetella organisms. Pooled real-time PCRs were used to detect B. pertussis and B. parapertussis based on the IS481 and IS1001 targets separately. RESULTS: Of the 629 subjects, 2 (0.3%) and 30 (4.8%) were confirmed to be culture-positive and PCR-positive, respectively, for B. pertussis, and 1 (0.2%) and 13 (2.1%) were confirmed to be culture-positive and PCR-positive, respectively, for B. parapertussis. All culture-positive samples were also PCR-positive. Furthermore, positive B. pertussis and B. parapertussis samples were found in all counties. CONCLUSIONS: Our results indicate that asymptomatic B. pertussis infections are common in school children in China, and asymptomatic B. parapertussis infections are more prevalent than previously documented.

[Study on the molecular typing and epidemiology of non-polio enterovirus in Yunnan Province, China during 2006-2010].

In order to explore the genotype distribution and molecular evolution of non-polio enterovirus (NPEVs)in Yunnan Province,the People's Republic of China, we sequenced and analyzed the partial VP1 coding region of 105 NPEVs isolated from acute flaccid paralysis (AFP) surveillance in Yunnan province during a 5- year study period from 2006 to 2010. The viral genomes of 105 NPEVs were translated to corresponding amino acid sequences and compared with those of the prototype strains, and the phylogenetic tree was constructed among these VP1 nucleotide sequences and other prototype strains from GenBank. Analysis showed that 18 isolates were classified into 7 serotypes of human enterovirus A species, while 77 isolates into 22 serotypes of B and 10 isolates into 4 serotypes of C species. However, we did not isolate any viruses which belonged to human enterovirus D species. Thus, under AFP surveillance, human enterovirus B species accounted for 73. 3% of the 105 isolates and was considered as the predominant one,followed by human enterovirus A(17. 1%) and human enterovirus C(9. 5%). Phylogenetic analysis showed that various serotypes of the virus and the corresponding prototype strains or other representative strains clustered into the same grooup, however, Yunnan strains and prototype strains were located in the different branches (except CA2,EV90 and EV76). The degree of variation was different even among the same genotype strains. This report showed that different genotype strains spread widely in Yunnan Province.

Gene chip array for differentiation of mycobacterial species and detection of drug resistance.

BACKGROUND: Gene chip array can differentiate isolated mycobacterial strains using various mycobacterium specific probes simultaneously. Gene chip array can evaluate drug resistance to isoniazid and rifampin of tuberculosis strains by detecting drug resistance related gene mutation. This technique has great potential for clinical application. We performed a retrospective study to investigate the capability of gene chip array in the rapid differentiation of species and detection of drug resistance in mycobacterium, and to evaluate its clinical efficacy. METHODS: We selected 39 patients (54 clinical mycobacterium isolates), used gene chip array to identify the species of these isolates and detect drug resistance to isoniazid and rifampin in Mycobacterium tuberculosis isolates. Meanwhile, these patients' clinical data were analyzed retrospectively. RESULTS: Among these 39 patients whose mycobacterium culture were positive, 32 patients' isolates were identified as Mycobacterium tuberculosis, all of them were clinical infection. Seven patients' isolates were identified as non-tuberculosis mycobacterium. Analyzed with their clinical data, only two patients were considered as clinical infection, both of them were diagnosed as hematogenous disseminated Mycobacterium introcellulare infection. The other five patients' isolates were of no clinical significance; their clinical samples were all respiratory specimens. Clinical manifestations of tuberculosis and non-tuberculous mycobacterial infections were similar. Isoniazid resistance was detected in two tuberculosis patients, while rifampin resistance was detected in one tuberculosis patient; there was another patient whose Mycobacterium tuberculosis isolate was resistant to both isoniazid and rifampin (belongs to multidrug resistance tuberculosis). The fact that this patient did not respond to routine anti-tuberculosis chemotherapy also confirmed this result. CONCLUSIONS: Gene chip array may be a simple, rapid, and reliable method for the identification of most mycobacterial species and detection of drug resistance in Mycobacterium tuberculosis. It is useful in diagnosis, treatment, and hospital infection control of mycobacterial infections, and it may have a great potential for clinical application.

[Molecular typing of enteroviruses from healthy children in the border areas of Yunnan Province and Myanmar and the genetic characteristics of ECHO7 and ECHO13 in 2009].

To explore the enteroviruses surveillance among healthy children under 15 years old in the border areas of Yunnan Province and Myanmar in 2009. The stool samples were collected from the healthy children under 15 years old who came from the border areas of Myanmar and Yunnan Province, virus isolation and sequencing were conducted for all the 271 samples. 6 strains of polioviruses (PVs) were detected from 271 stools with an isolation rate of 2.8%, which belonged to vaccine strains and 24 non-polioviruses (NPVs) were detected with an isolation rate of 8.9%. 24 NPVs belonged to human enterovirus group B (HEV-B) with 6 serotypes, HEV-A, HEV-C and HEV-D viruses were not isolated. Among them, 13 NPVs were E7 (54.17%) and 5 NPVs were E13 (20.83%). Our results showed that the enterovirus carrying rate in the border areas of Yunnan province was higher than the rate of routine acute flaccid paralysis (AFP) detection system. The HEV-B viruses were the only enteroviruses isolated. The phylogenetical analysis showed that Echovirus 7(E7) and 13 (E13) exhibited genetic polymorphism.