To determine the etiologic agent of an outbreak of influenza viruses from Changsha Foothill Mountain International School in 2009, and to analyze the HA Gene Characteristic of the H1N1 influenza viruses. Twenty-five nasopharyngeal swab specimens from the outbreak of influenza viruses were tested by conventional RT-PCR and influenza viruses isolated simultaneously. Virus isolated (A/Yuelu/314/2009) from the outbreak was sequenced by CEQTM 8000 Genetic Analysis System and the sequencing results submitted to GenBank (Accession No: FJ912843), then the sequencing data was analyzed by ClustalX and Mega4.1softwares. Results showed the influenza viruses A(H1N1) of positive were 18 cases by influenza viruses isolated tests and 21 cases by conventional RT-PCR, respectively. The nucleotide and amino acid sequence homology of the HA gene of A/Yuelu/314/2009 are 99% compare with the vaccine strain (A/Brisbane/59/2007) in 2008~2009 years. The HA sequence data also showed that had 6 amino acid mutations (V148A, S158N, G202A, I203D, A206T, W435R), and the S158N located at antigenic site B of HA protein. Nine potential glycosylation sites (27, 28, 40, 71, 151, 176, 303, 497, 536) in the HA sequence of A/Yuelu/314/2009 is the same with A/Brisbane/59/2007, and the sequences of potential glycosylation sites were conserved. In this study, laboratory evidence diagnosed seasonal influenza A virus (H1N1) as the etiologic agent of the outbreak. The virus isolated (A/Yuelu/314/2009) strain of H1N1 subtype is not a new variant in Changsha in 2009 compare with the vaccine strain (A/Brisbane/59/2007), the outbreak of influenza A virus (H1N1) from Changsha Foothill Mountain International School maybe are caused by the change in genetic characteristics between vaccine strains and the decreased of immunity to influenza A virus (H1N1) in the crowd.

OBJECTIVE:To investigate the effect of shen-fu injection on toxic reaction relief in the chemotherapy for moderate to advanced non-small cell lung cancer(NSCLC).METHODS:130 patients of NSCLC were randomly divided into treatment group and control group,2 groups both received the second-generation regimen for chemotherapy,the major chemotherapeutic agents included vinorelbine,gemcitabine and paclitaxel,the treatment group was given intravenous shen-fu injection 60ml/d for continuous2weeks plus chemotherapy.RESULTS:The toxic reactions in treatment group significantly decreased compared to that in the control group(P

Abstract: Objective　To investigate the molecular characteristic and evolutionary trends of full-genome sequences of coxsackievirus A2 (CV-A2) and A5 (CV-A5) in Changsha City. Methods　The CV-A2 and CV-A5 strains were isolated and detected from patients with hand, foot and mouth disease (HFMD) cases. The full-genome sequences of CV-A2 and CV-A5 strains were obtained using NGS sequencing. Homology and phylogenetic tree analysis were performed, and the recombination regions of the strains were examined by SimPlot software. Results　The full-genome sequences of CV-A2 and CV-A5 strains were obtained from routine surveillance cases of HFMD in Changsha in 2019. The CV-A2 strain was named S281/Changsha/CHN/2019 with the full-genome sequence of 7 422 bp long; the CV-A5 strain was named S272/Changsha/CHN/2019 with the full-genome sequence of 7 425 bp long. Homology analysis of the isolates by comparison with the nucleic acid sequences of CV-A2 and other CV-A2 strains in China showed that the non-structural protein region shared lower similarity than that of structural protein region. The CV-A2 showed 79.20% similarity with Fleetwood strain (NC038306), showed the highest similarity 95.60% with MN419014 strain from Hubei Province. The non-structural protein 3C and 3D region shared the lowest similarity with MN419014, 90.51 and 92.06%, respectively. Phylogenetic tree analysis showed that 3C and 3D regions were located in the CV-A4 branch. Amino acid mutation sites were found in non-structural protein region, and the amino acid sequence in structural protein region was conserved. SimPlot analysis showed that genetic recombination was found in the 3C and 3D region of CV-A2 strains. The full-genome sequence of CV-A5 showed 80.7% similarity with the Swartz (AY421763) and 97.43% similarity with the strain (MH111030) from Australian. Homology analysis showed that the non-structural protein region shared lower similarity than that of structural protein region, based on full-genome of CV-A5. Phylogenetic tree analysis showed that CV-A5 and MH111030 were in the same branch, indicating that CV-A5 strain not from local. The amino acid sequence of CV-A5 strain was conserved. Conclusions　The CV-A2 strain in Changsha City shared genome sequence information with CV-A4, and the CV-A5 strain was imported from abroad. Our findings are expected to understand the molecular and recombination characteristics of CV-A2 and CV-A5, provided the data of evolution and genetic features of the coxsackievirus, and interrupt disease transmission in a timely and effective manner.

OBJECTIVETo identify the self-preparation monoclonal antibody which target to clenbuterol, and set up the standard curve to clenbuterol (CL) detection.

METHODSThe affinity constants and activity of the monoclonal antibody which target to CL were determined by ELISA. ELISA was also used to confirm whether the monoclonal antibody had any across-reaction with BSA and CL analogues. The rat ascites which contains the monoclonal antibody target to CL was purified by (NH4)2SO4 salt-out method and further by affinity column. At last, the CL detection standard curve which based on indirect competition ELISA was established.

RESULTSThe ELISA experiment showed that the antibody titer was 10(6) and the monoclonal antibody affinity constants was 2.90 x 10(10) L/mol. The result of the indirect competition ELISA confirmed that the monoclonal antibody had no cross-reaction with BSA and a few kind of CL analogue. CL detection standard curve based on indirect competition ELISA was established, which R2 was 0.9812, and the lowest detectable limit was 1.0 ng/ml.

CONCLUSIONThe standard curve based on indirectly competitioning ELISA was established. The self-preparation monoclonal antibody which target to CL has high affinity and high specific to CL, which had established the foundation to the advanced development of the CL immune test paper and CL ELISA kit.

Animals ; Antibodies, Monoclonal ; chemistry ; isolation & purification ; Antibody Affinity ; Clenbuterol ; immunology ; Cross Reactions ; Enzyme-Linked Immunosorbent Assay ; Limit of Detection ; Rats

Child ; China ; epidemiology ; Genes, Viral ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; virology

OBJECTIVETo study the characteristics of the chest X-ray images in children infected with enterovirus 71.

METHODSA total of 120 children with enterovirus 71 infection between April, 2010 and July, 2011 were classified into three groups according to the disease condition: mild (31 cases), severe (43 cases) and life-threatening (46 cases). The period from the onset of clinical symptoms to the first chest X-ray imaging examination and the results of the first chest X-ray findings were compared among the three groups.

RESULTSThe period from the onset of clinical symptoms to the first chest X-ray imaging examination in the mild, severe and life-threatening groups was 26-48 hrs (median 37 hrs), 10-36 h (median 23 hrs) and 2-36 hrs (median 19 hrs) respectively. Chest X-ray abnormalities were initially observed at 30 hrs after the onset of clinical symptoms in the mild group, at 23 hrs in the severe group and at 2 hrs in the life-threatening group (P<0.01). The mild group presented an initial imaging abnormality rate of 5.8%, the severe group 81.3% and the life-threatening group 100%. The life-threatening group showed a significantly higher initial X-ray abnormality rate than the other two groups (P<0.01). In terms of chest X-ray performance, the mild group usually presented lung marking thickening or vagueness. Most children in the severe group presented lung effusion and consolidation. Signs of pulmonary edema were found in the life-threatening group, and lesions in the life-threatening group were characterized by wide distribution and many lung lobe involvements.

CONCLUSIONSThe interval between the onset of clinical symptoms and the initial chest X-ray examination, the period of time of, and the onset of clinical symptoms, at which chest X-ray abnormalities, the abnormality rate and the severity of chest X-ray findings may be paralleled to the clinical situation in children with enterovirus 71 infection.

Child, Preschool ; Enterovirus A, Human ; Enterovirus Infections ; diagnostic imaging ; Female ; Humans ; Infant ; Male ; Radiography, Thoracic

OBJECTIVETo compare the clinical efficacy in the treatment of allergic rhinitis (AR) of lung qi deficiency and cold syndrome between Jin's three-needle therapy and western medication.

METHODSSixty-six patients were randomized into an acupuncture group and a western medication group, 33 cases in each one. In the acupuncture group, acupuncture was applied at three-nose points [Yingxiang (LI 20), Shangyingxiang (EX-HN 8) and Yintang (GV 29); Cuanzhu (BL 2) was added for frontal headache] and three-back points [Dazhu (BL 11), Fengmen (BL 12) and Feishu (BL 13)], once every day. Ten treatments made one session. Two sessions of treatment were required. In the western medication group, desloratadine oral suspension was prescribed, 5 mg each time, once a day, for 20 days. The scores of the symptoms and physical signs in AR patients as well as the clinical efficacy were observed between the two groups.

RESULTSThe total effective rate was 93.9% (31/33) in the acupuncture group, which was better than 72.7% (24/33) in the western medication group (P < 0.05). After treatment, the scores of AR symptoms and physical signs as well as the total score were all reduced compared with those before treatment in the two groups (all P < 0.01). The score of every item in the acupuncture group was lower than that in the western medication group after treatment (score of symptoms: 4.70 +/- 2.07 vs 6.55 +/- 2. 69, score of physical signs: 0.85 +/- 0.67 vs 1.45 +/- +0.62, total score: 5.36 +/- 2.70 vs 8.00 +/- 2.91, all P < 0.01).

CONCLUSIONJin's three-needle therapy achieves superior efficacy on AR of lung-qi deficiency and cold syndrome, which is better than desloratadine oral suspension.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Adolescent ; Adult ; Child ; Female ; Humans ; Lung ; physiopathology ; Male ; Middle Aged ; Needles ; Qi ; Rhinitis, Allergic ; Rhinitis, Allergic, Perennial ; physiopathology ; therapy ; Young Adult

To develop a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for rapid and sensitive detection of Norwalk GII. 4 primers which recognized 6 distinct regions on the RNA-dependent RNA polymerase gene of Norwalk GII were designed and used for LAMP assay. Norwalk GII RNA was amplified under isothermal conditions (65 degrees C) for 120 min, and LAMP results were then judged with naked eye, SYBR Green I staining, electrophoretic analysis and restriction digestion. To evaluate the specificity of the RT-LAMP, 48 fecal specimens of Norwalk GII and 12 fecal specimens of group A rotaviruses were tested. To compare the sensitivity of the RT-LAMP with that of conventional RT-PCR, Norwalk GII RNA was serially diluted and amplified by RT-LAMP and RT-PCR, respectively. With 46 fecal specimens of Norwalk GII, observation with naked eyes, SYBR Green I staining and electrophoretic analysis were able to detect the PCR products in the RT-LAMP assay. The specificity of RT-LAMP products was also confirmed by digestion of the RT-LAMP products with restriction enzymes. No RNA amplification was observed in 2 fecal specimens of Norwalk GII and 12 fecal specimens of group A rotaviruses. The specificity of the RT-LAMP assay with regard to RT-PCR were 100% for Norwalk GII. The detection limits of RT-LAMP was 15.6 pg/tube for Norwalk GII and similar to that of a RT-PCR assay. Compared to RT-PCR, the RT-LAMP assay has been proven to be a rapid, sensitive, specific and accurate method for detection of the Norwalk GII in fecal specimens, and that RT-LAMP assay is potentially useful for the rapid detection of Norwalk GII from fecal specimens in outbreaks of infectious diarrhea.
Caliciviridae Infections ; virology ; Feces ; virology ; Humans ; Norwalk virus ; genetics ; isolation & purification ; RNA Replicase ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viral Proteins ; genetics

OBJECTIVETo investigate the indications, safety and difficulties of one stage thyroidectomy and bilateral neck lymph node dissection for well-differentiated thyroid carcinoma.

METHODSA retrospective review was carried out in 36 well-differentiated thyroid carcinoma patients so treated from 1990 to 2004. Various incisions including H, L and modified Kocher types were selected according to the location of primary tumor and status of cervical lymph node metastasis. Either total thyroidectomy or sub-total thyroidectomy combined with bilateral neck lymph node dissection according to the principles of modified radical neck lymph node dissection: preserving the internal jugular vein, spinal accessory nerve and sternocleidomastoid muscles.

RESULTSThere was no operative death in this group. Postoperative complications included: 2 wound bleeding, 3 recurrent laryngeal nerve resection due to tumor involvement, 1 recurrent laryngeal nerve injury, 2 unilateral internal branch of superior laryngeal nerve injury, 9 unilateral external branch of superior laryngeal nerve injury, 3 unilateral accessory nerve injury, 5 unilateral sympathetic nerve injury, 2 unilateral phrenic nerve injury, 6 chylus fistula, 13 temporary hypoparathyroidism, 2 permanent hypoparathyroidism. The dissected lymph nodes were found to be positive from 0 to 21 in each patient with a mean of 8.3. Of the 36 patients: 31 had bilateral positive lymph nodes; 3 unilateral positive; 2 bilateral negative lymph nodes. The follow up period ranged from 1 to 13 years, Three patients died of distant metastasis, 1 died of cerebral vascular accident. 7 patients lost in follow-up. Totally, 25 patients are still alive, 3 patients had local relapse and were surgically treated again.

CONCLUSIONThe procedure of one-stage thyroidectomy and bilateral neck lymph node dissection for well-differentiated thyroid carcinoma is safe, as it is mandatory that at least one unilateral internal jugular vein should be preserved; one unilateral recurrent laryngeal nerves and accessory nerves should not be injured. Well-differentiated thyroid carcinoma patients whose bilateral cervical lymph nodes are clinically suspected to be positive (obviously enlarged, hard, purplish grapelike lymph node) or are confirmed pathologically to be positive are indications for one-stage thyroidectomy and bilateral neck lymph node dissection. Total or sub-total thyroidectomy should be undertaken with emphasis that at least one parathyroid with blood supply should be preserved. It is of utmost importance that not only the cancer be completely resected but the function of the organs be preserved.

Adenocarcinoma, Follicular ; secondary ; surgery ; Adult ; Carcinoma, Papillary ; secondary ; surgery ; Female ; Follow-Up Studies ; Humans ; Hypothyroidism ; etiology ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neck Dissection ; Neoplasm Recurrence, Local ; Postoperative Complications ; Retrospective Studies ; Survival Rate ; Thyroid Neoplasms ; pathology ; surgery ; Thyroidectomy ; methods

BACKGROUNDA high mortality rate of pancreatic cancer becomes a bottleneck for further treatment with long-term efficacy. It is urgent to find a new mean to predict the early onset of pancreatic cancer accurately. The authors hypothesized that genetic variants of cationic trypsinogen (PRSS1) gene could affect trypsin expression/function and result in abnormal activation of protease activated receptor-2 (PAR-2), then lead to pancreatic cancer. The aim of this study was to elaborate some novel mutations of PRSS1 gene in the patients with pancreatic cancer.

METHODSTotally 156 patients with pancreatic cancer and 220 unrelated individuals as controls were enrolled in this study. The mutations of PRSS1 gene were analyzed by direct sequencing. K-ras Mutation Detection Kit was used to find the general k-ras gene disorder in the pancreatic cancer tissue. Then the clinical data were collected and analyzed simultaneously.

RESULTSThere were two patients who carried novel mutations which was IVS 3 + 157 G > C of PRSS1 gene in peripheral blood specimens and pancreatic cancer tissue. What's more, it was surprising to find a novel complicated mutation of exon 3 in PRSS1 gene (c.409 A > G and c.416 C > T) in another young patient. The complicated mutation made No. 135 and No. 137 amino acid transfer from Thr to Ala and Thr to Met respectively. No any mutation was found in the normal controls while no mutations of k-ras gene were detected in the three patients.

CONCLUSIONMutations of PRSS1 gene may be an important factor of pancreatic cancer.

Adult ; Asian Continental Ancestry Group ; Female ; Humans ; Male ; Mutation ; Pancreatic Neoplasms ; genetics ; Trypsin ; genetics