Effects of scalp acupuncture plus acupuncture exercise therapy on walking ability in children with spastic cerebral palsy / 针灸推拿医学（英文版）

Objective: To compare the effect of scalp acupuncture and scalp acupuncture plus acupuncture exercise therapy (AET) on walking ability in children with spastic cerebral palsy (CP). Methods: A total of 60 spastic CP children with gross motor function classification system (GMFCS) grades Ⅰ-Ⅲ were divided into a control group and an observation group by the random number table method, with 30 cases in each group. Both groups were treated with the same conventional rehabilitation and scalp acupuncture therapy for CP. The control group received conventional rehabilitation first and then scalp acupuncture. The observation group received AET, which was to receive the conventional rehabilitation and scalp acupuncture simultaneously. Before and after treatment, the clinical efficacy was evaluated by the modified Ashworth scale (MAS) score, scores of dimensions D and E of the gross motor function measure (GMFM) scale, walking speed, and walking distance. Results: During treatment, there were 2 dropouts in the observation group. After 3 courses of treatment, the MAS scores in both the control group and observation group decreased compared with the same group before treatment (P<0.05), and the scores of dimensions D and E of the GMFM, walking speed, and walking distance were increased (P<0.05); the between-group comparison showed that the MAS score in the observation group was lower than that in the control group (P<0.05), and the scores of dimensions D and E of the GMFM, walking speed, and walking distance in the observation group were higher or longer than those in the control group (P<0.05). Conclusion: W ith the same treatments, scalp acupuncture combined with AET is superior to the conventional scalp acupuncture method in reducing lower-limb muscle tone, improving standing balance ability, and walking stability in children with spastic CP.

Challenges and considerations on quality control and evaluation of pathogen metagenomic next-generation sequencing / 生物工程学报

Metagenomic next-generation sequencing (mNGS) could be used for pathogen detection from nearly all types of clinical samples. Especially, the unique diagnostic capability of pathogen mNGS detecting unknown causative agent of infectious diseases makes this method become an importation complement and irreplaceable component for conventional routine laboratory test. However, the complexity of the testing process, the rapid product update, and the insufficiency in quality control and evaluation methods that all make clinical transformation, industry development, and regulation of this technology full of challenge and uncertainty. This review briefly introduces the technical advantages and challenges, and describes the general workflow and quality control steps in details. Finally, it focuses on current considerations regarding quality evaluation methods and standards for pathogen mNGS.

Molecular detection and genotyping of human papillomavirus / 中华肿瘤杂志

The development of cervical cancer is associated with persistent infection of high-risk human papillomavirus. The detection and genotyping of HPV could be used to evaluate the epidemiology of HPV infections, monitor HPV vaccine efficacy, and screen cervical cancer. There are a lot of commercially available molecular tests for HPV, based on different methodologies and detection systems. In principle, it mainly includes two categories, namely signal amplification and target amplification. Most of them are based on PCR amplification, such as fluorescent PCR, PCR-reverse hybridization, etc. The performances of detection reagents are different. In addition, HPV genotyping assays based on next-generation sequencing and quantitative HPV detection kits are developed. However, only a small number of commercial assays have been clinically verified. A large number of assays which may bring greater values in the screening of cervical cancer are needed to be clinically validated.

The investigation and analysis of college students' awareness, cognition and usage of shared courses on the internet / 中华医学教育探索杂志

Objective To investigate college students' awareness and cognition of 84 shared courses on the Internet and analyze the discrepancies between different ethnicity and majors in Xinjiang Medical University. Methods Purposive sampling was adopted with questionnaire of 1 448 students of 26 classes in Xinjiang Medical University. The survey aimed to investigate students' awareness and cognition of these In- ternet courses. Obtained data were statistically analyzed using SPSS 13.0, and the test level α=0.05. Result The hits of preventative medicine, Chinese medicine, clinical medicine and pharmacy were 19.0%, 33.3%, 35.7% and 11.9% respectively. Among the courses whose hits exceeded 50,000, clinical medicine accounted for 50%, preventive medicine for 21.4%, Chinese medicine and pharmacy for 14.3% respectively, there being no significant difference in overall hits (P>0.05). 90.1% of the surveyed students knew shared courses on the Internet, there being no difference between gender, ethnicity and profession (P>0.05). 89.8% had an accurate understanding of the concept of shared courses on the Internet, there being differences between gender and major ( χ2gender=11.013, P=0.026; χ2major=136.08, P=0.000) without significant differences in ethnicity ( χ2=11.378, P=0.497). 71.9% of the students used shared courses on the Internet as reference resources for the course study, 46.6% for the learning content, 35.5%for the understanding of other resources related to the course, 30.1% for lab-class and exercises, 10% for discussion on online course forum. The Han and Kazak students used the courses as a study, experiment and exercise tool, while the Uyghur and Kazak mainly used these for learning and discussion on the forum, there being differences in ethnicity ( χ2=26.889, P=0.001); the usage rate of the courses of preventive medicine students is higher, whereas pharmacy students relatively low, there being significant differences in major ( χ2=38.01, P=0.004). The Kazakh and Uygur students mainly used the courses to formulate learning plans, and the Han students to learn the current curriculum. 44.7% of preventive medicine students used the courses to improve their abilities, which was significantly higher than those of other majors. Only 18.4% and 1% of pharmacy students used the courses to formulate learning plans and to improve their abilities, which were both lower than those of other majors, showing ethnic and professional differences ( χ2ethnic=37.654, P=0.001; χ2major=73.68, P=0.000). Conclusion Students' awareness of shared courses on the Internet is high and their cognition is accurate. However, there are differences in the ways and purposes of the employment of the courses between different ethnicity and majors. The main reason may be related to major, the quality of the courses, as well as the lack of effec-tive supervision and evaluation system, suggesting that the management department in our university should strengthen the supervision and evaluation of the courses and give full play to the important role of shared courses on the Internet.

Situation and suggestions on IVD industrial standards / 中国医疗器械杂志

This paper briefly introduces the working procedure of in vitro diagnostic products (IVD) industrial standards, and elaborates the importance of professional standards for production and supervision. Based on the analysis of working progress during the past 10 years, some problems and countermeasures on project setting, participation, standard material, personnel training, work cycle are put forward, which are helpful for the future development of the IVD.

Study of the relationship between antibody level and protection induced by human papillomavirus(HPV) L2-peptide / 中华微生物学和免疫学杂志

Objective To detect the protection induced by HPV-58 L2 11-200 AA in animal, and analyze the relationship between antibody or neutralizing antibody titers and the protection generated by the immunizmg agent. Methods The peptide of HPV-58 L2 11-200 AA was expressed in E. coli and the mice were immunized with the peptide after purification and adsorption with aluminum adjuvant. The protection provided by different immunizing doses was detected in the mouse model against the challenge of the pseud-ovirions of human papiilomavirus types 58. The total antibodies and neutralizing antibody titers of serum were tested with ELISA and neutralization assay against HPV-58 pseudovirus, respectively. The total antibodies or neutralizing antibody titers that can protect the mouse from infection were analyzed. Results The mice can be protected from the challenge with HPV pseudovirus when the immunizing dose was 8 μg. The neutralizing antibody can not be detected in the immune serum by neutralization assay against pseudovirus. The total anti-body level has a corresponding relationship with the protection showed in mouse model. The results of total antibodies detected by ELISA showed that when the titer of total antibodies was ≥25 000, luminescent signal can not be detected and the mice can be protected from pseudovirus infection. Conclusion HPV-58 L2 11-200 AA peptide can protect mice from pseudovirus infection. L2 peptide has a promising perspective to be a candidate vaccine and the level of total antibodies in the immune serum can be used as a surrogate for the evaluation of protection against HPV infection.

Evaluation of cellular immune responses in mice elicited by Chinese AIDS candidate vaccines / 中华微生物学和免疫学杂志

Objective To compare cellular immune responses in mice elicited by Chinese different AIDS candidate vaccines.Methods According to their different immunization procedures,BALB/c mice were immunized with 6 AIDS candidate vaccines,separately.Spleen cells were isolated for the detection of cellular immune response to HIV-specific peptides using enzyme-linked immunosorbent spot(ELISPOT)assay and intracellular cytokine staining(ICS)method.Results AIDS vaccines were evaluated by using potential T-cell epitopes(PTE)Gag,Env and Pol peptides pool and ELISPOT.The positive conversion rates for cellular immune response of 1#-6# vaccines fluctuated from 70% to 100%.The vaccine-induced cellular immune responses to specific peptides pool are different not only in magnitude but also in breadth.The Th1type cytokines,IFN-γand IL-2,were detected with ELISPOT in 1# and 2# vaccines.The productions of IFN-γand IL-2 induced by both of the two vaccines showed a moderate correlation(r1 =0.62,P1 ＜0.01 ;r2=0.79,P2 ＜ 0.01).The positive conversion rate of IFN-γ secreting cells of 1 # vaccine was 66.7%(10/15)mice detected with both ELISPOT and ICS.And the results tested by ELISPOT and ICS showed moderate correlation(r = 0.55,P ＜ 0.05).Conclusion The magnitude and breadth of cellular immune responses induced by different AIDS candidate vaccines are different.Being induced by different AIDS candidate vaccines,the IFN-γand other Th1 type cytokines detected by ELISPOT or ICS could be used to evaluate the cellular immune responses in mice.

Detection of HPV Types and Neutralizing Antibodies in Women with Genital Warts in Tianjin City,China / 中国病毒学·英文版

The serum samples and corresponding cervical swabs were collected fi'om 50 women with genital warts from Tianjin city,China.The neutralizing antibodies against HPV-16,-18,-58,-45,-6 and-11 in serum samples were tested by using pseudovirus-based neutralization assays and HPV DNAs in cervical swabs were also tested by using a typing kit that can detect 21 types of HPV.The results revealed that 36%(18/50)of sera were positive for type-specific neutralizing antibodies with a titer range of 160-2560,of which 22%(11/50),12%(6/50),10%(5/50),4%(2/50),4%(2/50)and 2%(1/50)were against HPVs-6,-16,-18,-58,.45 and-11,respectively.Additionally,60%(30/50)of samples were HPV DNA-positive,in which the most common types detected were HPV-68(18%),HPV-16(14%),HPV-58(12%),HPV-33(8%)and HPV-6,HPV-11,HPV-18 and HPV-52(6% each).The concordance between HPV DNA and corresponding neutralizing antibodies was 56%(28/50)with a significant difference(P＜0.05).The full-length sequences of five HPV types(HPV-42,-52,-53,-58 and-68)were determined and exhibited 98%-100% identities with their reported genomes.The present data may have utility for investigating the natural history of HPV infection and promote the development of HPV vaccines.

The impact of site-directed deglycosylation of the HIV-1 envelope (Env) on its immunogenicity and assembly of functional pseudovirus / 中华微生物学和免疫学杂志

Objective To study the influence of site-directed deglycosylation of the HIV-1 envelope (Env) on its immunogenicity and assembly of functional pseudovirus. Methods Site-directed deglycosylation were performed using cycling mutagenesis and selection of mutants with DpnⅠ. Single-cycle infection assay was employed to analyze the effect of the mutations on the ability of functional pseudovirus assembly. The influence of deglycosylations on the immunodeficiency of Env was evaluated using pseudovirusbased neutralization assay and ELISPOT assay. Results Mutant N197Q induced higher neutralization activities for both pseudoviruses, but lower Env-specific T-cell response. And N197Q rendered the Env to lose the ability of functional pseudovirus assembly. Mutant G2 induced higher neutralization activities for pseudovirus 74-2 but lower for pseudovirus Wt, and had almost no influence on Env-specific T-cell response and functional pseudovirus forming. Conclusion The site-directed deglycosylation of the HIV-1 Env affects the pseudovirus forming and its immunogenicity.

Establishment and primary application of pseudovirus-based neutralization assay for HIV-1 / 中华微生物学和免疫学杂志

Objective To establish a pseudovirus-based neutralization assay. Methods The functional gp160 genes were amplified from plasmids containing HIV-1 gene. The products were ligased into pcDNA3.1 plasmid and positive clones were screened by digestion with restriction enzymes. The pseudoviruses were harvested by co-transfection of the positive clone and pSG3△env plasmid. The neutralizing activities of monoclonal antibodies and HIV-1 antibody positive plasma were measured by these pseudoviruses. Results The four strains of psedoviruses (CHB01, CHB02, CHBC03 and CHAE04) had been successfully obtained. Monoclonal antibody 4E10 could neutralize all of 4 pseudoviruses while 2G12 could not neutralize any pesudoviruses. Monoclonal antibody 2F5 could neutralize pseudovirus CHB01, CHB02 and CHAE04 but not CHBC03, while IgG1b12 could neutralize pseudovirus CHB01, CHB02 and CHBC03 but not CHAE04. The neutralizing activities of 43 of HIV-1 antibody positive plasma against different subtypes of pseudovirus were significant differences and the cross-neutralization effects for some samples exist. Conclusion The harvested pseudoviruses could be used in the neutralization assay. However, the neutralizing characteristics of different pseudoviruses may be different.

Preliminary Study on a Potential Panel for Quality Assurance of ELISPOT / 中国病毒学·英文版

The ELISPOT assay is increasingly used for assessing cellular immune responses in clinical trials of HIV-1 or cancer vaccines. However, to date, data from clinical trials do not consistently show that immune responses are correlated with clinical endpoints. This is due in part to the lack of assay standardization and validation across laboratories and therefore, a quality control panel is required to establish competency and comparability amongst different laboratories. In this study peripheral blood mononuclear cells (PBMCs) from healthy individuals were screened and frozen in liquid nitrogen. The recovery and viability of the PBMCs and the frequencies of interferon (IFN)-γ-secreting cells after CEF peptide pool stimulation were detected after various intervals in seven different laboratories. The recovery and viability did not differ significantly after different intervals. Although the frequencies of IFN (interferon)-γ-secreting cells among thawed PBMCs (peripheral blood mononuclear cells) fluctuated after CEF peptide pool stimulation at different intervals, they were not significantly decreased compared with those among fresh PBMCs. However, the viabilities, recoveries and frequencies of IFN-γ-secreting cells differed significantly among the seven laboratories. Our results indicate that cryopreserved PBMCs could be used as a quality control panel for ELISPOT. However, the procedures for ELISPOT need to be standardized amongst different laboratories.

Comparison of the Immunogenicities of HIV-1 Mutants Based on Structural Modification of env / 中国病毒学·英文版

Eleven env mutants were designed and generated by site-directed mutagenesis of the regions around Nab epitopes and deletions of variable regions in env.The immunogenicities of the generated mutants were evaluated using single-cycle infection neutralization assays with two pseudoviruses and IFN-γELISPOT.Overall,five mutants(dWt,M2,M5-2,M5-1 and dM7)induced highed neutralization activities for both pseudoviruses than plasmid Wt,while only two of the mutants(dWt and M5-2)showed significant differences(P<0.05).Two mutants(M2 and dM2)induced more Env-specific T cells than plasmid Wt.Statistically however,significance was only reached for mutant M2.Thus,properly modified HIV-1 Env may have the potential to induce potent cellular and humoral immune responses.

Application of promoting standing training for children with cerebral palsy of the spastic type / 中国组织工程研究

Totally 20 children with cerebral palsy of spastic type, aged 1.5-5.0 years, who were treated in Department of Children Nerve Rehabilitation, Nanhai Maternity and Kid Hospital Affiliated to Guangzhou University of Traditional Chinese Medicine, were selected from March to December 2005. Of them, 11 cases were moderate degree of cerebral palsy and 9 cases were severe. The diagnostic criterion of cerebral palsy referred to the diagnostic criterion established at the Countrywide Special Topic Proseminar of Cerebral Palsy in 2004. The method to diagnose cerebral palsy of spastic type referred to the method of Cerebral Palsy Academy of United States in 1956. Cerebral palsy was clinically characterized by the very brisk reflex of extending. The degree was graded by mild, moderate and severe according to the paralyzed part, mental development, speech development, activity of daily living and complication. The children were promoted to stand and walk by themselves mainly with standing promoting training and with restraining method as complement. The course of treatment was 6 months. The curative effect was evaluated by the abducent angle of the hip joint, the angle of popliteal fossa, the dorsiflexion angle of ankle, and the forth item of gross motor function measure, which were measured before and after the treatment. Theresult showed that the range of motion of 20 children was improved. The score of gross motor function measure of standing item improved evidently. A total of 8 cases had significant effect and 12 cases had valid effect. The total effective rate was 100%.