[Clinical characteristics and prognosis of 12 cases of lupus nephritis complicated with thrombotic microangiopathy]. / 12ä¾‹ç‹¼ç–®æ€§è‚¾ç‚Žåˆå¹¶è¡€æ “æ€§å¾®è¡€ç®¡ç— å˜ä¸´åºŠç‰¹ç‚¹åŠé¢„åŽåˆ†æž.

OBJECTIVES: To investigate the clinical characteristics, pathological features, treatment regimen, and prognosis of children with lupus nephritis (LN) and thrombotic microangiopathy (TMA), as well as the treatment outcome of these children and the clinical and pathological differences between LN children with TMA and those without TMA. METHODS: A retrospective analysis was conducted on 12 children with LN and TMA (TMA group) who were admitted to the Department of Nephrology, Children's Hospital of Nanjing Medical University, from December 2010 to December 2021. Twenty-four LN children without TMA who underwent renal biopsy during the same period were included as the non-TMA group. The two groups were compared in terms of clinical manifestations, laboratory examination results, and pathological results. RESULTS: Among the 12 children with TMA, 8 (67%) had hypertension and 3 (25%) progressed to stage 5 chronic kidney disease. Compared with the non-TMA group, the TMA group had more severe tubulointerstitial damage, a higher Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) score at onset, and higher cholesterol levels (P<0.05). There were no significant differences between the two groups in the percentage of crescent bodies and the levels of hemoglobin and platelets (P>0.05). CONCLUSIONS: There is a higher proportion of individuals with hypertension among the children with LN and TMA, as well as more severe tubulointerstitial damage. These children have a higher SLEDAI score and a higher cholesterol level.

A prognostic model based on DNA methylation-related gene expression for predicting overall survival in hepatocellular carcinoma.

Background: Hepatocellular carcinoma (HCC) continues to increase in morbidity and mortality among all types of cancer. DNA methylation, an important epigenetic modification, is associated with cancer occurrence and progression. The objective of this study was to establish a model based on DNA methylation risk scores for identifying new potential therapeutic targets in HCC and preventing cancer progression. Methods: Transcriptomic, clinical, and DNA methylation data on 374 tumor tissues and 50 adjacent normal tissues were downloaded from The Cancer Genome Atlas-Liver Hepatocellular Carcinoma database. The gene expression profiles of the GSE54236 liver cancer dataset, which contains data on 161 liver tissue samples, were obtained from the Gene Expression Omnibus database. We analyzed the relationship between DNA methylation and gene expression levels after identifying the differentially methylated and expressed genes. Then, we developed and validated a risk score model based on the DNA methylation-driven genes. A tissue array consisting of 30 human hepatocellular carcinoma samples and adjacent normal tissues was used to assess the protein and mRNA expression levels of the marker genes by immunohistochemistry and qRT-PCR, respectively. Results: Three methylation-related differential genes were identified in our study: GLS, MEX3B, and GNA14. The results revealed that their DNA methylation levels were negatively correlated with local gene expression regulation. The gene methylation levels correlated strongly with the prognosis of patients with liver cancer. This was confirmed by qRT-PCR and immunohistochemical verification of the expression of these genes or proteins in tumors and adjacent tissues. These results revealed the relationship between the level of relevant gene methylation and the prognosis of patients with liver cancer as well as the underlying cellular and biological mechanisms. This allows our gene signature to provide more accurate and appropriate predictions for clinical applications. Conclusion: Through bioinformatics analysis and experimental validation, we obtained three DNA methylation marker: GLS, MEX3B, and GNA14. This helps to predict the prognosis and may be a potential therapeutic target for HCC patients.

Evaluation of a new frequency-volume chart for children with primary monosymptomatic nocturnal enuresis: a prospective, comparative study.

INTRODUCTION: To improve compliance with voiding diaries in children with primary monosymptomatic nocturnal enuresis (PMNE), a new modified 3-day weekend frequency-volume chart (FVC) was designed, and the compliance and validity of this modified FVC was evaluated by comparing with the International Children's Continence Society (ICCS) recommended voiding diary. METHODS: A total of 1200 patients with PMNE were enrolled in the study from 13 centers in China and were randomly assigned to record this modified FVC or the ICCS-recommended voiding diary. The primary outcome measure was the compliance, assessed by comparing the completing index and the quality score of diaries between two groups. The secondary outcome measure was the validity, evaluated by comparing the constituent of subtypes, micturition parameters and response rate to desmopressin. RESULTS: Among the 1200 participants enrolled in the study, 447 patients completed the ICCS-recommended voiding diary and 469 completed the modified diary. The diurnal completing index and the quality score of the modified FVC group were better than those of the ICCS group. In addition, there was no significant difference between these two groups in the subtype classification, or in the response rate to desmopressin. CONCLUSIONS: The modified FVC could be applied to obtain the voiding characteristics of children with PMNE as the ICCS-recommended voiding diary does and offers a reasonable and better choice for children with PMNE from the unselected population in the future.

Effects of lower body positive pressure treadmill on functional improvement in knee osteoarthritis: A randomized clinical trial study.

BACKGROUND: Knee joint pain and stiffness are the two main symptoms of knee osteoarthritis (OA) and thus restrict a patient's activities, such as walking and walking up and downstairs. The lower body positive pressure (LBPP) treadmill as one of the emerging body weight support system devices brings new hope for exercise-related rehabilitation for knee OA patients. AIM: To investigate the biomechanical effects and the subjective clinical assessment of LBPP treadmill walking exercise when compared with conventional therapy in mild to moderate knee OA patients. METHODS: Eighteen patients with mild-to-moderate knee OA were recruited in this randomized controlled trial (RCT) study. The eligible knee OA patients were randomly assigned to two groups: LBPP and control groups. The patients in the LBPP group performed an LBPP walking training program for 30 min/session per day, 6 d per week for 2 wk whereas the patients in the control group performed walking on the ground for the same amount. All patients underwent clinical assessments and three-dimensional gait analysis at pre- and 2-wk post-treatment. RESULTS: The Western Ontario and McMaster Universities Arthritis Index and visual analog scale scores in both the LBPP group and control group were found to decrease significantly at the post-treatment point than the pre-treatment point (LBPP: 70.25 ± 13.93 vs 40.50 ± 11.86; 3.88 ± 0.99 vs 1.63 ± 0.52; control: 69.20 ± 8.88 vs 48.10 ± 8.67; 3.80 ± 0.79 vs 2.60 ± 0.70, P < 0.001). Moreover, compared with the control group, the LBPP group showed more improvements in walking speed (P = 0.007), stride length (P = 0.037), and knee range of motion (P = 0.048) during walking, which represented more improvement in walking ability. CONCLUSION: The results of our RCT study showed that the LBPP group has a greater effect on improving gait parameters than the conventional group, although there was no significant advantage in clinical assessment. This finding indicates that LBPP treadmill walking training might be an effective approach for alleviating pain symptoms and improving lower extremity locomotion in mild to moderate knee OA patients.

Lipidation of polyethylenimine-based polyplex increases serum stability of bioengineered RNAi agents and offers more consistent tumoral gene knockdown in vivo.

Recently we have established a novel approach to produce bioengineered noncoding RNA agents (BERAs) in living cells that carry target RNAi molecules (e.g., siRNA and miRNA) and thus act as "prodrugs". Using GFP-siRNA-loaded BERA (BERA/GFP-siRNA) as a model molecule, this study was to define the in vitro and in vivo knockdown efficiency of BERAs delivered by liposome-polyethylenimine nanocomplex (lipopolyplex or LPP). Compared to in vivo-jetPEI® (IVJ-PEI) and polyplex formulations, LPP offered greater protection of BERA/GFP-siRNA against degradation by serum RNases. Particle sizes and zeta potentials of LPP nanocomplex remained stable over 28 days when stored at 4 °C. Furthermore, comparable levels of BERA/GFP-siRNA were delivered by LPP and IVJ-PEI to luciferase/GFP-expressing human SK-Hep1-Luc-GFP or A549-Luc-GFP cells, which were selectively processed into target GFP-siRNA and subsequently knocked down GFP mRNA and protein levels. In addition, LPP-carried BERA/GFP-siRNA was successfully delivered into xenograft tumors and offered more consistent knockdown of tumoral GFP mRNA level in an orthotopic hepatocellular carcinoma (HCC) SK-Hep1-Luc-GFP xenograft mouse model, while IVJ-PEI formulation showed larger variation. These findings demonstrated that lipidation of polyplexes improved serum stability of biologic RNAi molecules, which was efficiently delivered to orthotopic HCC tissues to knock down target gene expression.

Combination therapy with bioengineered miR-34a prodrug and doxorubicin synergistically suppresses osteosarcoma growth.

Osteosarcoma (OS) is the most common form of primary malignant bone tumor and prevalent among children and young adults. Recently we have established a novel approach to bioengineering large quantity of microRNA-34a (miR-34a) prodrug for miRNA replacement therapy. This study is to evaluate combination treatment with miR-34a prodrug and doxorubicin, which may synergistically suppress human OS cell growth via RNA interference and DNA intercalation. Synergistic effects were indeed obvious between miR-34a prodrug and doxorubicin for the suppression of OS cell proliferation, as defined by Chou-Talalay method. The strongest antiproliferative synergism was achieved when both agents were administered simultaneously to the cells at early stage, which was associated with much greater degrees of late apoptosis, necrosis, and G2 cell cycle arrest. Alteration of OS cellular processes and invasion capacity was linked to the reduction of protein levels of miR-34a targeted (proto-)oncogenes including SIRT1, c-MET, and CDK6. Moreover, orthotopic OS xenograft tumor growth was repressed to a significantly greater degree in mouse models when miR-34a prodrug and doxorubicin were co-administered intravenously. In addition, multiple doses of miR-34a prodrug and doxorubicin had no or minimal effects on mouse blood chemistry profiles. The results demonstrate that combination of doxorubicin chemotherapy and miR-34a replacement therapy produces synergistic antiproliferative effects and it is more effective than monotherapy in suppressing OS xenograft tumor growth. These findings support the development of mechanism-based combination therapy to combat OS and bioengineered miR-34a prodrug represents a new natural miRNA agent.

Bioengineering Novel Chimeric microRNA-34a for Prodrug Cancer Therapy: High-Yield Expression and Purification, and Structural and Functional Characterization.

Development of anticancer treatments based on microRNA (miRNA/miR) such as miR-34a replacement therapy is limited to the use of synthetic RNAs with artificial modifications. Herein, we present a new approach to a high-yield and large-scale biosynthesis, in Escherichia coli using transfer RNA (tRNA) scaffold, of chimeric miR-34a agent, which may act as a prodrug for anticancer therapy. The recombinant tRNA fusion pre-miR-34a (tRNA/mir-34a) was quickly purified to a high degree of homogeneity (>98%) using anion-exchange fast protein liquid chromatography, whose primary sequence and post-transcriptional modifications were directly characterized by mass spectrometric analyses. Chimeric tRNA/mir-34a showed a favorable cellular stability while it was degradable by several ribonucleases. Deep sequencing and quantitative real-time polymerase chain reaction studies revealed that tRNA-carried pre-miR-34a was precisely processed to mature miR-34a within human carcinoma cells, and the same tRNA fragments were produced from tRNA/mir-34a and the control tRNA scaffold (tRNA/MSA). Consequently, tRNA/mir-34a inhibited the proliferation of various types of human carcinoma cells in a dose-dependent manner and to a much greater degree than the control tRNA/MSA, which was mechanistically attributable to the reduction of miR-34a target genes. Furthermore, tRNA/mir-34a significantly suppressed the growth of human non-small-cell lung cancer A549 and hepatocarcinoma HepG2 xenograft tumors in mice, compared with the same dose of tRNA/MSA. In addition, recombinant tRNA/mir-34a had no or minimal effect on blood chemistry and interleukin-6 level in mouse models, suggesting that recombinant RNAs were well tolerated. These findings provoke a conversation on producing biologic miRNAs to perform miRNA actions, and point toward a new direction in developing miRNA-based therapies.

Chimeric MicroRNA-1291 Biosynthesized Efficiently in Escherichia coli Is Effective to Reduce Target Gene Expression in Human Carcinoma Cells and Improve Chemosensitivity.

In contrast to the growing interests in studying noncoding RNAs (ncRNAs) such as microRNA (miRNA or miR) pharmacoepigenetics, there is a lack of efficient means to cost effectively produce large quantities of natural miRNA agents. Our recent efforts led to a successful production of chimeric pre-miR-27b in bacteria using a transfer RNA (tRNA)-based recombinant RNA technology, but at very low expression levels. Herein, we present a high-yield expression of chimeric pre-miR-1291 in common Escherichia coli strains using the same tRNA scaffold. The tRNA fusion pre-miR-1291 (tRNA/mir-1291) was then purified to high homogeneity using affinity chromatography, whose primary sequence and post-transcriptional modifications were directly characterized by mass spectrometric analyses. Chimeric tRNA/mir-1291 was readily processed to mature miR-1291 in human carcinoma MCF-7 and PANC-1 cells. Consequently, recombinant tRNA/mir-1291 reduced the protein levels of miR-1291 target genes, including ABCC1, FOXA2, and MeCP2, as compared with cells transfected with the same doses of control methionyl-tRNA scaffold with a sephadex aptamer (tRNA/MSA). In addition, tRNA-carried pre-miR-1291 suppressed the growth of MCF-7 and PANC-1 cells in a dose-dependent manner, and significantly enhanced the sensitivity of ABCC1-overexpressing PANC-1 cells to doxorubicin. These results indicate that recombinant miR-1291 agent is effective in the modulation of target gene expression and chemosensitivity, which may provide insights into high-yield bioengineering of new ncRNA agents for pharmacoepigenetics research.

A general approach to high-yield biosynthesis of chimeric RNAs bearing various types of functional small RNAs for broad applications.

RNA research and therapy relies primarily on synthetic RNAs. We employed recombinant RNA technology toward large-scale production of pre-miRNA agents in bacteria, but found the majority of target RNAs were not or negligibly expressed. We thus developed a novel strategy to achieve consistent high-yield biosynthesis of chimeric RNAs carrying various small RNAs (e.g. miRNAs, siRNAs and RNA aptamers), which was based upon an optimal noncoding RNA scaffold (OnRS) derived from tRNA fusion pre-miR-34a (tRNA/mir-34a). Multi-milligrams of chimeric RNAs (e.g. OnRS/miR-124, OnRS/GFP-siRNA, OnRS/Neg (scrambled RNA) and OnRS/MGA (malachite green aptamer)) were readily obtained from 1 l bacterial culture. Deep sequencing analyses revealed that mature miR-124 and target GFP-siRNA were selectively released from chimeric RNAs in human cells. Consequently, OnRS/miR-124 was active in suppressing miR-124 target gene expression and controlling cellular processes, and OnRS/GFP-siRNA was effective in knocking down GFP mRNA levels and fluorescent intensity in ES-2/GFP cells and GFP-transgenic mice. Furthermore, the OnRS/MGA sensor offered a specific strong fluorescence upon binding MG, which was utilized as label-free substrate to accurately determine serum RNase activities in pancreatic cancer patients. These results demonstrate that OnRS-based bioengineering is a common, robust and versatile strategy to assemble various types of small RNAs for broad applications.

Hydroa vacciniforme-like cutaneous T-cell lymphoma.

A 14-year-old Chinese girl had a 6-year history of recurrent lesions on her head, face, and limbs. Epstein-Barr virus (EBV)-IgM was positive. Histopathological findings revealed focal lymphocyte invasion in subcutaneous panniculus adiposus, mainly surrounding the blood vessels. Immunohistochemistry showed CD3+, CD4+, CD5+, CD8+, TIA-1+, GrB+, CD56-, and L26-. In situ hybridization staining for EBV-encoded small nonpolyadenylated RNA (EBER)-1 was positive. The patient showed significant improvement in clinical symptoms after being treated with acyclovir and IFN-α in this patient.

CYP2C9*3(1075A > C), ABCB1 and SLCO1B1 genetic polymorphisms and gender are determinants of inter-subject variability in pitavastatin pharmacokinetics.

A pharmacokinetics study was conducted in 12 Chinese volunteers following a single dose of 1 mg, 2 mg and 4 mg of pitavastatin calcium in an open-label, randomized, three-period crossover design. Plasma concentrations of pitavastatin acid and pitavastatin lactone were determined by a HPLC method. Single-nucleotide polymorphisms (SNPs) in ABCB1, ABCG2, SLCO1B1, CYP2C9 and CYP3A5 were determined by TaqMan (MGB) genotyping assay. An analysis was performed on the relationship between the aforementioned SNPs and dose-normalized (based on 1 mg) area under the plasma concentration-time curve extrapolated to infinity [AUC(0-infinity)] and peak plasma concentration (Cmax) values of the acid and lactone forms of pitavastatin. Pitavastatin exhibited linear pharmacokinetics and great inter-subject variability. Compared to CYP2C9*1/*1 carriers, CYP2C9*1/*3 carriers had higher AUC(0-infinity) and Cmax of pitavastatin acid and AUC(0-infinity) of pitavastatin lactone (P<0.05). With respect to ABCB1 G2677T/A, non-G carriers had higher Cmax and AUC(0-infinity) of pitavastatin acid, and Cmax of pitavastatin lactone compared to GT, GA or GG genotype carriers (P<0.05). Gene-dose effects of SLCO1B1 c.521T> C and g.11187G > A on pharmacokinetics of the acid and lactone forms were observed. Compared to non-SLCO1B1*17 carriers, SLCO1B1*17 carriers had higher Cmax and AUC(0-infinity) of the acid and lactone forms (P<0.05). Significant sex difference was observed for pharmacokinetics of the lactone. Female SLCO1B1 521TT subjects had higher Cmax and AUC(0-infinity) of pitavastatin lactone compared to male 521TT subjects, however, such gender difference disappeared in 521 TC and 521CC subjects. Pitavastatin pharmacokinetics was not significantly affected by ABCB1 C1236T, ABCB1C3435T, CYP3A5*3, ABCG2 c.34G > A, c.421C > A, SLCO1B1 c.388A>G, c.571T>C and c.597C>T. We conclude that CYP2C9*3, ABCB1 G2677T/A, SLCO1B1 c.521T>C, SLCO1B1 g.11187G > A, SLCO1B1*17 and gender contribute to inter-subject variability in pitavastatin pharmacokinetics. Personalized medicine should be necessary for hypercholesterolaemic patients receiving pitavastatin.

An overview of ABC and SLC drug transporter gene regulation.

Membrane transporters play a significant role in drug absorption, distribution and excretion, and they consequently affect the pharmacokinetics, efficacy and safety of a drug. Under certain circumstances, such as pathological processes or exposure to certain substances, the expression of drug transporters is modified in cells. Change in transporter expression and function may affect cellular drug disposition resulting in different drug responses. This raises a number of questions such as which drugs are likely to modulate the expression of drug transporters, what factors support this process, and which transporters are influenced in a particular situation. In this paper, we summarize recent findings to find an answer to these questions. Particularly, we present an overview of the transcription factors involved in the regulation of a given drug transporter, the signaling transduction pathways that contribute to drug transporter gene expression, and xenobiotics and endobiotics that initiate the processes.

[Etiology survey on virus of acute respiratory infection in Guangzhou from 2006 to 2009].

OBJECTIVE: To investigate the pathogens of acute respiratory infection (ARI) in Guangzhou from 2006 to 2009. METHODS: A total of 1554 cases of ARI patients in Second Affiliated Hospital of Guangzhou Medical College from September 2006 to September 2009, were recruited in the survey. The sample of throat and pharyngeal swab were collected from each patient.11 types of virus including influenza A (FluA), influenza B (FluB), adenovirus (ADV), human rhinovirus (HRV), respiratory syncytial virus (RSV), parainfluenza virus type 1, type 2, type 3 (HPIV1, HPIV2, HPIV3), human metapneumovirus (MPV) and human coronavirus (HCoV) type 229E, type OC43 were detected by Fluorescence Quota PCR method. The epidemic feature and clinical characteristic of each virus were then analyzed. RESULTS: Virus were found in 1024 samples in total, accounting for 65.9% (1024/1554). RSV was the most common virus, which was found in 261 samples (16.8%); and followed by HRV as 13.9% (216/1554), FluA as 11.6% (181/1554), MPV as 6.5% (101/1554), FluB as 6.4% (99/1554), HPIV as 4.9% (76/1554), ADV as 3.5% (55/1554) and HCoV as 2.3% (35/1554). HPIV and HCoV shared a similar infection ratio among different age groups. The infection ratio of FluA and FluB was highest among 15-24 years old group, accounting for 16.5% (29/176) and 7.4% (13/176) respectively. MPV, RSV and HRV were the main pathogens caused infection among children under 4 years old, accounting for 9.7% (49/503), 21.7% (109/503) and 18.9% (95/503). The infection ratio of ADV was 6.0% (19/318), which was the most common pathogen among 5-14 years old patients. The incidence rate of HPIV and HRV showed no obvious seasonal features; while the prevalence of FluA, FluB, RSV, ADV, MPV and HCoV changed significantly in different seasons.22.2% (227/1024) ARI patients co-infected other respiratory virus.90.1% (163/181) FluA patients, 88.9% (88/99) FluB patients and 92.7% (51/55) ADV patients had high fever symptoms. CONCLUSION: RSV was the main pathogen of ARI, and the new-found virus MPV was also another crucial pathogen. Some pathogens' incidence rate were related to the season and patient's age. Co-infections of other respiratory virus were also detected in parts of ARI patients.

[Research progress of zebrafish used in drug metabolism].

Zebrafish is widely used as a model organism in the process of drug discovery. It expresses drug metabolizing enzymes like cytochrome P450 (CYP450), uridine 5'-diphospho-glucuronosyltransferase (UGT) and nuclear receptors like pregnane X receptor (PXR), aryl hydrocarbon receptor (AHR), etc. This article summarized the profiles of main drug metabolizing enzymes and nuclear receptors, and reviewed the advances on xenobiotics metabolism in zebrafish.

Is acquired symmetrical acrokeratoderma a new dermatosis? Two case reports and Chinese literature review.

BACKGROUND: A series of cases characterized by symmetrical acral hyperkeratosis, mainly involving the dorsal surface of the hands, feet, and wrists, but sparing the palmoplantar areas, as well as rapid immersion upon exposure to water have been recently described in China, but similar disorders have not been reported in the English literature. METHODS: The clinicopathologic features of two cases of acquired symmetrical acrokeratoderma were reported and 27 Chinese patients were reviewed. RESULTS: The disorder typically occurred in young or middle aged men. Brown to black hyperkeratotic patches were symmetrically distributed particularly on the wrists and dorsum of hands, fingers, and feet, but without involvement of palms and soles. The lesions became whitish with mild swelling immediately after contact with water and improved generally in winter. Histopathologic examination revealed epidermal hyperkeratosis, acanthosis, and papillomatous hyperplasia, as well as superficial perivascular lymphohistiocytic infiltrate. Main ultrastructural features of the immersed lesion were epidermal hyperkeratosis and spongiosis with partial split of the desmosomes. CONCLUSION: This disorder may be a new dermatosis, and the term "acquired symmetrical acrokeratoderma" could exactly reflect its clinicopathologic characteristics.

[Molecular epidemiological characteristics of norovirus gastroenteritis outbreaks in Guangdong from 2005 to 2008].

To study the molecular epidemiological characteristics of norovirus gastroenteritis outbreaks in Guangdong, we collected fecal and anal swabs specimens from 24 outbreaks of acute gastroenteritis from 2005 to 2008 to detect norovirus. Specimens were detected by RT-PCR and then sequenced. The descriptive data were also collected. According to our research, 19 of 24 outbreaks of gastroenteritis were positive for norovirus. The occurrence time was from October to next February mainly. The strains in 2005 belonged to G II-3 genotype and all outbreaks occurred in kindergarten and school. But from autumn of 2006, the outbreaks were all caused by G II-4/2006b variant and occurred in universities and community. The number of outbreaks in 2007 increased greatly and covered all over province. The nucleotide sequences of Guangdong strains in some sites showed high regional identity. Our results showed that with the shift of genotype from G II-3 to G II-4, occurrence of norovirus outbreaks increased greatly. The outbreaks of norovirus caused by G II-4/2006b variant spreaded widely and the involved population covered children and adult, indicating the strong invasiveness of this variant.

[Investigation of human metapneumovirus in children with acute respiratory tract infections in Guangzhou areas].

OBJECTIVE: To find out the status of human metapneumovirus (hMPV) infection in children under 14 years old with acute respiratory tract infections (ARI) in Guangzhou, analyze the epidemiology and clinical characteristics among the hMPV-infected children, and provide some basis for research of hMPV. METHODS: All 521 throat and pharyngeal swabs were collected among the children with acute respiratory tract infections in outpatient departments and those admitted to the wards from September 2006 to August 2008. Then total nucleic acid was extracted from respiratory specimens. The 213 nucleosides of nucleoprotein gene were detected by RT-PCR and 16 strong positive samples were picked to compare with the sequence of hMPV in GenBank after the sequence of the amplification products were determined. Then applied statistical analysis to the data of the collected patients. RESULTS: All 521 samples were detected by RT-PCR, and confirmed that N gene was positive in 39 samples with a detection rate of 7.49%, and the peak time was in October and April. The 16 amplification products were compared by using the analysis of gene sequence. The nucleocapsid protein (N) gene similarity to BJ1897 of Beijing was up to 99%, and to AY550156 of Thailand was up to 97%, genotype B was the most common genotype. CONCLUSION: There existed hMPV infection in children acute respiratory system diseases in Guangzhou areas, in which the children under the age of 6 years were accounted for the main group, however there was no difference in gender. The main symptoms of the patients with hMPV infection were high fever and cough symptom of catarrh. Co-infections other than respiratory virus with hMPV were detected as 41.03% of positive samples.

Multiple outbreaks of acute hemorrhagic conjunctivitis due to a variant of coxsackievirus A24: Guangdong, China, 2007.

Acute hemorrhagic conjunctivitis (AHC) is usually caused by enterovirus 70, coxsackievirus A24(CA24v) and adenoviruses. Several outbreaks of AHC caused by a CA24v have occurred since it was imported into China in 1971. Multiple outbreaks of AHC reappeared in 10 cities of Guangdong during June to November in 2007. The epidemic began in the June, and spread extensively, with a peak in the September. A total of 31,659 cases were reported to center for disease control and prevention of Guangdong, it was estimated that the number of actual AHC was >200 thousands. Forty conjunctival swab specimens were collected from the cases diagnosed clinically with AHC. (RT)-PCR testing on these conjunctival specimens revealed the presence of an enterovirus, and this was confirmed by 16 isolates. We demonstrated the most likely etiological agent for the multiple outbreaks was a variant of coxsackievirus A24 by molecular typing using a partial VP1 sequence. Sequence comparison and phylogenetic analyses of the VP1 and 3Cpro gene regions were performed by Neighbor-joining method, the strains from different outbreaks and different geographical areas within Guangdong had no sequence divergence in 2007. The representative isolates from mainland of China including Hangzhou, Ningbo, Beijing, Yunnan, Liaoning, and Henan were analyzed in this study. Phylogenetic analysis revealed theses isolates were located in different clusters, a close phylogenetic and chronological relationship with Singaporean, South Korean and Thailand isolates had been observed. This confirms CA24v circulated in China's mainland has not evolved independently, but co-evolved with the isolates of Southeast Asia.

[Characteristics, evolution and variation of M genes of human avian H5N1 strains in Guangdong].

In order to reveal variation and evolution of M genes of human avian H5N1 influenza strains, the M genes of human avian H5N1 strains in Guangdong were sequenced and the M genes of global strains were searched out from Internet. They were analyzed by DNAStar 5. 0 and their revolutionary speeds were studied by means of combining the epidemiological data. It was found that M1 genes of 53 H5N1 strains and M2 genes of 51 strains during 1997-2006 were homologously classified into two groups: the strains from Hong Kong during 1997 (G I) were one group and the strains from Hong Kong, Vietnam, Thailand, Indonesia, China mainland, Turkey, Iraq, Azerbaijan, Egypt during 2003-2006 (G II ) were the another group. There were 20 substitutions of amino acids in M1 gene of all strains (7.94%, 20/252), where there were 9 amino acids in strains during 2003-2006 differing from the strains in 1997, meanwhile there were 22 substitutions of amino acids in M2 gene of all strains (22.7%, 22/97), where there were 4 amino acids in strains during 2003-2006 differing from the strains in 1997. In the synonymous variation, Ks values in M1 were 26.8 x 10(-6)-42.6 x 10(-6) Nt/d, and Ka values 4.39 x 10(-6)-6.98 x 10(-6) Nt/d, where there was more rapid speed of synonymous substitution than that of replacement, which showed that there existed less human immunological pressure and negative selective pressure by biological test. Ks values in M2 were 13.1 x 10(-6)-23.4 x 10(-6) Nt/ d, and Ka values 9.1 x 10(-6)-16.2 x 10(-6) Nt/d; where the ratios of Ks to Ka was 1.0-1.6 times as there was the neutral selective pressure in TL-676-05 strain. There was an amino acid substitution of S224, N in M1 gene of strains during 2003-2006 and an increas in a glycoprotein domain NSS224-226. The secondary structure of M2 protein varied as the substitution of C50 F of eight strains from Indonesia in 2005. The strains G I did not reemerge after Hong Kong human avian H5N1 influenza event. An increase of a glycoprotein domain NSS224-226 in M1 protein during 2003-2006 might be related with virus pathogenicity. Human avian H5N1 influenza M gene evolved frequently in nature, which might have an impact on its capacity of human-to-human transmission.