Objective To establish a clinical test assay for detecting common respiratory viruses and enteroviruses (EV) by using mixed cultured Madin-Darby canine kidney cells (MDCK), human epidermoid cancer cells (Hep-2) and African green monkey kidney cells (Vero) to isolate common respiratory viruses and enteroviruses. Methods Throat swabs with influenza A and B viruses,adenovirus and EV71 were incubated with mixed cultured MDCK, Hep-2 and Vero in a single vial to observe the presence of cytopathic effects. Polymerase chain reaction (PCR), reverse transcription (RT)-PCR and monoclonal antibody-based immunofluorescene assay were also used for confirmatory test. Results The sensitive cell lines developed obvious cytopathic effects to the corresponding viruses, which were confirmed by the specific green particles observed by immunofluorescence assay and specific target PCR segments. Conclusions The shell-vial of mixed cells can simultaneously isolate different common respiratory viruses and EV. The isolated pathogens can be further confirmed by antigen test and PCR. This assay may improve the diagnosis of clinical viral diseases.

Objective To understand the sensitivity of cytopathogenic effect (CPE) in MadinDarby canine kidney cells(MDCK) that cultured influenza A pharyngeal swab specimens of patients for one,two and three passages. Methods Influenza A pharyngeal swab specimens of patients were inoculated in MDCK for three blind passages. The presence of CPE of every passage was observed by inverted microscope. Results Of the 279 influenza A pharyngeal swab specimens of patients tested by colloidal gold, the presence of CPE in MDCK for one,two and three passages was 65.9%(184/279),91.4%(255/279) and 96.4%(269/279), respectively. Two hundred and seventy-one of 279specimens were identified as influenza A by multiplex reverse transcription-polymerase chain reaction (RT-PCR). Conclusion The positive separation rate can reach more than 95% by inoculating influenza A pharyngeal swab specimens of patients in MDCK for three blind passages.

This article referred to the relevant guidelines for vaccine clinical trials and the accumulated experience in the development and review of human rabies vaccines, as well as combined the technical recommendations for human rabies vaccines from major health facilities such as WHO, to discuss the key considerations in the design of clinical trials for human rabies vaccines, including control vaccines, evaluation of efficacy endpoints, clinical subjects, the sample size, immunity persistence, re-exposure immunization schedule and post-marketing study. This article aimed to provide reference for the design of human rabies vaccine clinical trials.

β-thalassemia is a single-gene genetic disease caused by β globin gene mutations leading to the fact that red blood cells are unable to form normal adult hemoglobin, and then patients develop hemolytic anemia. Current treatment regimens mainly include allogenetic hematologic stem cell transplantation, symptomatic regular blood transfusions and the use of iron removers to reduce iron load. Some severe patients have quite poor prognoses and deadly consequences if not treated timely. Genetically modified autohematopoietic stem cells can provide a new treatment option for patients with β thalassemia, which may achieve a long-term and stable increase in hemoglobin level through a single dose, making one-time cure β-thalassemia possible. This paper reviews the key elements of clinical trial design for β-thalassemia gene therapy from the aspects of efficacy evaluation endpoints, clinical trial design, enrollment population, and subject monitoring in order to provide a reference for pharma-therapeutic research and development enterprises.

Objective To investigate the changes and related factors of maternal thyroid autoantibodies during early pregnancy. Methods Urinary iodine concentration( UIC) , serum thyroid stimulating hormone( TSH) , free thyroxine ( FT4 ) , thyroid-peroxidase antibody ( TPOAb ) , thyroglobulin antibody ( TgAb ) concentrations were determined in 7 190 women during early pregnancy in an iodine-sufficient region of China. Results The prevalence of TPOAb positivity and TgAb positivity were 8. 7% and 12. 0% respectively. The prevalence of overt hypothyroidism and subclinical hypothyroidism increased significantly in group of thyroid antibody positivity. The prevalence of TPOAb positivity and TgAb positivity presented a U-shaped curve, ranging from mild iodine deficiency to iodine excess, especially increased significantly in the group with UIC<100 μg/L. Conclusion Prevalence of thyroid antibodies positivity became higher during early pregnancy. The positive thyroid autoantibodies during pregnancy were significantly associated with maternal hypothyroidism. Both iodine excess and iodine deficiency are risk factors of positive thyroid antibodies.

Chimeric antigen receptor (CAR) T lymphocyte has shown attractive prospects in the treatment of lymphohematopoietic malignancies including B-cell lymphoblastic leukemia, B-cell lymphoma and multiple myeloma. Many applicants have submitted investigational new drug (IND) applications to Center for Drug Evaluation of National Medical Products Ggency, however, many of the INDs have problems in patient selection, prognostic indicators and risk management, etc, which might hinder the evaluation of the safety and efficacy of CAR-T cells. Thus, we made some suggestions on the above-mentioned problems through summarizing clinical experience and communicating with domestic clinical experts, which the sponsors and researchers can refer to when conducting CAR-T cell clinical trials for registration.

Objective: To analyze vaccination situation of oral live attenuated rotavirus vaccine (LLR strain) among children from six provinces in China. Methods: In 2014, we selected 12 counties in Guangdong, Jiangsu, Chongqing, Jiangxi, Heilongjiang and Gansu provinces by using stratified cluster random sampling method and extract information of children born from January 1, 2008 to December 31, 2012 from Children's Immunization Information System. We investigated ten children of each birth cohort in each county by checking the vaccination certification, and a total of 606 children were investigated. A survey was conducted to check the information of the children's vaccination certification with the data of Children's Immunization Information System by questionnaire including the basic information (province, county, name, gender, birth date, etc) and the rotavirus vaccination (vaccination date, dose, etc) to analyze the rotavirus vaccination situation. Results: 340 of 606 children were male. There were 121, 124, 122, 119 and 120 children born in 2008-2012, respectively. The proportions of the first and the second dose of rotavirus vaccination were 32.8% (199) and 9.7% (59). The proportion of the third dose of rotavirus vaccination among children born between 2008 and 2010 was 3.5% (13) since children born in 2011 and 2012 did not reach the age of third dose vaccination. The proportion of the first dose of rotavirus vaccination in high, middle and low per capita disposable income areas was 45.0% (91), 37.7% (77) and 15.5% (31) respectively (χ²= 43.15, P<0.001). Among 199 children vaccinated with the first dose of vaccine, the vaccination age mainly concentrated in 2 to 21 months, of which the peak was 5 to 13 months (66.8%, 133). The intervals between 2 doses of vaccination were mainly from 12 to 13 months (42.4%, 25) among the 59 children who received at least 2 doses of vaccine. In the 13 children vaccinated with 3 doses, the intervals between the second and the third dose were 12 months (5). Of the 271 doses of rotavirus vaccine vaccinated during 2008-2014, 34.7% (94 doses) were vaccinated in June-August, 88 were vaccinated simultaneously with 18 other vaccines, accounting for 32.5% of the total. Of the 18 other vaccines, inactivated vaccines such as diphtheria vaccine (30 doses), Hib vaccine (14 doses), group A meningitis vaccine (10 doses) were predominant. Conclusion The proportion of rotavirus vaccination was low and the vaccination age was relatively late. The vaccination mode was different from the recommendation of WHO. It is recommended that routine immunization of rotavirus vaccines should be carried out in early-months of children.