Development of a sandwich ELISA for detecting 3AB non-structural protein of foot-and-mouth disease virus / 生物工程学报

Antigenic purity is important for quality control of the foot-and-mouth (FMD) whole virus inactivated vaccine. The recommended method for evaluation the antigenic purity of FMD vaccine is to check the serum conversion to non-structural protein (NSP) 3AB antibody after 2 to 3 times inoculation of animals with inactivated vaccine. In this study, we developed a quantitative ELISA to detect the amount of residual 3AB in vaccine antigen, to provide a reference to evaluate the antigenic purity of FMD vaccine. Monoclonal antibody (Mab) of NSP 3A and HRP-conjugated Mab of NSP 3B were used to establish a sandwich ELISA to quantify the NSP 3AB in vaccine antigen of FMD. Purified NSP 3AB expressed in Escherichia coli was serially diluted and detected to draw the standard curve. The detectable limit was determined to be the lowest concentration of standard where the ratio of its OD value to OD blank well was not less than 2.0. Results: The OD value was linearly corelated with the concentration of 3AB protein within the range between 4.7 and 600 ng/mL. The correlation coefficient R² is greater than 0.99, and the lowest detectable limit is 4.7 ng/mL. The amount of 3AB protein in non-purified inactivated virus antigen was detected between 9.3 and 200 ng/mL depending on the 12 different virus strains, whereas the amount of 3AB in purified virus antigen was below the lowest detectable limit. The amount of 3AB in 9 batches of commercial FMD vaccine antigens was between 9.0 and 74 ng/mL, whereas it was below the detectable limit in other 24 batches of commercial vaccine antigens. Conclusion: the sandwich ELISA established in this study is specific and sensitive to detect the content of 3AB protein in vaccine antigen of FMD, which will be a useful method for evaluation of the antigenic purity and quality control of FMD inactivated vaccine.

Meta-analysis of Risk Factors of Linezolid-induced Thrombocytopenia / 中国药房

OBJECTIVE： To evaluate risk factors of linezolid-induced thrombocytopenia systematically， and to provide reference for rational drug use in clinic. METHODS： Retrieved from PubMed， Embase， Cochrane library， Web of Science， CBM， CNKI and Wanfang database， during database establishment to Oct. 2018， clinical studies about risk factors of linezolid-induced thrombocytopenia were collected， and the data of literatures met criteria were collected. After Newcastle-Ottawa scale （NOS） was applied for evaluating the quality of included literatures. Meta-analysis was conducted by using Rev Man 5.3 software. RESULTS： Sixteen clinical studies involving 2 264 patients in total were included. Results of Meta-analysis showed that daily per kg dose （DKPD） [SMD＝0.62， 95％CI（0.29，0.95）， P＝0.000 2]， low platelet count before medication [SMD＝－0.90， 95％CI（－1.62， －0.18）， P＝0.01]， low creatinine clearance rate [SMD＝－0.65， 95％CI（－1.10，－0.19）， P＝0.005]， long treatment course [SMD＝0.45， 95％CI（0.18，0.71）， P＝0.000 9]， low body weight  [SMD＝－0.36， 95％CI（－0.60，－0.11），P＝0.005] significantly influenced the occurrence of thrombocytopenia. CONCLUSIONS： The risk factors associated with linezolid-induced thrombocytopenia include low baseline platelet count， low creatinine clearance rate， low body weight， long medication course and high DKPD.

Correlation of sperm DNA fragmentation index with semen parameters / 中华男科学杂志

Objective@#To explore the relationship of sperm DNA fragmenation index (DFI) with semen parameters and assess its application value in the evaluation of semen quality.@*METHODS@#A total of 9 694 semen samples were collected and examined for sperm DFI and high DNA stainability (HDS) by flow cytometry-assisted sperm chromatin structure analysis (SCSA). According to the WHO Laboratory Manual for the Examination and Processing of Human Semen (5th Ed), the samples were divided into a normal group and abnormal groups A (sperm concentration ［SC］: ［11.3－14.0］ ×10⁶/ml, total sperm motility ［TSM］: 30%－39%, progressively motile sperm ［PMS］: 24%－31%), B (SC: ［7.5－11.2］ ×10⁶/ml, TSM: 20%－29%, PMS: 16%－23%), C (SC: ［3.8－7.4］ ×10⁶/ml, TSM: 10%－,19% PMS: 8%－15%) and D (SC: ［0－3.7］×10⁶/ml, TSM: 0－9%, PMS: 0－7%), and also into three sperm DFI groups (DFI 30%). The correlation between sperm DFI and seminal parameters was analyzed by Pearson correlation and multiple linear regression analyses.@*RESULTS@#DFI was dramatically lower in the normal than in the abnormal groups (P < 0.01), and increased in proportion to the decrease of semen parameters in the abnormal groups A, B, C and D (P < 0.01). Pearson correlation analysis showed that DFI was correlated positively with age (r = 0.15, P < 0.01), abstinence time (r = 0.10, P < 0.01), semen volume (r = 0.05, P < 0.01) and HDS (r = 0.15, P < 0.01), but negatively with semen pH (r = －0.06, P < 0.01), SC (r = －0.27, P < 0.01), TSM (r = －0.53, P < 0.01), PMS (r = －0.52, P < 0.01) and morphologically normal sperm (r = －0.16, P < 0.01). Multiple linear regression analysis revealed that TSM, SC, age, abstinence time and semen pH were five important variables associated with DFI, with standardized regression coefficients of －0.47, －0.19, 0.12, 0.07, and －0.04, respectively (all P < 0.01).@*CONCLUSIONS@#There is a moderate correlation between sperm DFI and semen parameters, which can be used synergistically for the assessment of semen quality.

Meta-analysis of Rituximab Injection Associated Pneumonia / 中国药房

OBJECTIVE: To systematically evaluate the relationship between rituximab and the occurence of pneumonia, and to provide evidenle-based reference in clinic. METHODS: Retrieved from PubMed, Embase, Cochrane Library, Web of Science, CBM, CNKI, Wanfang database and other database, RCTs, quasi-RCTs, controlled clinical trials were searched from inception to Mar. 2017. After literature screening, data extraction, literature quality evaluation and analysis of bias risk referring to Cochrane Intervention Measure System Evaluation Manual (5. 3 edition), Meta-analysis was conducted by using RevMan 5. 3 software. RESULTS: A total of 16 literatures were finally included, consisting of 14 RCTs and 1 386 patients. The results of Meta-analysis showed that the incidence of rituximab associated pneumonia in observation group was lower than control group, but without statistical significance [OR=1. 19, 95％ CI (0. 75, 1. 91), Z=0. 74, P=0. 46]. Subgroup analysis was conducted according to sample size, multiple-center, intervention dose, there was no statistical significance in the incidence of rituximab-associated pneumonia between 2 groups (P＞0. 05). CONCLUSIONS: There was no significant difference in the incidence of pneumonia between observation group and control group, rituximab is not associated with the occurrence of pneumonia. Due to methodology limit of included studies, most of high quality long-term follow-up studies have not been included. Large-scale and high quality clinical studies are required for further valuation.

Follow-up of children born after intracytoplasmic sperm injection with epididymal and testicular spermatozoa / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>To evaluate the safety of intracytoplasmic sperm injection (ICSI) with epididymal or testicular sperm, this study compared children born after ICSI treatment with epididymal or testicular sperm with children conceived after ICSI with ejaculated sperm.</p><p><b>METHODS</b>This retrospective study included 317 children born after ICSI with percutaneous epididymal sperm aspiration (PESA), 103 children born after ICSI with testicular sperm aspiration (TESA), and a control group of 1008 children born after ICSI with ejaculated sperm. All of the patients received their assisted reproductive treatment in the Reproductive Medicine Center of the First Affiliated Hospital of Zhengzhou University from January 2004 to December 2011. Data, such as the rate of stillbirths, perinatal mortality, gestational age, birth weight, and the rate of congenital malformations of the three groups, were compared.</p><p><b>RESULTS</b>PESA and TESA children were not different from ICSI children in the rate of stillbirths, perinatal mortality, infant mortality rate, gestational age, the rate of prematurity, and the rate of malformations (P > 0.05). A slight increase in birth defects was reported in the TESA group compared with those in the control group, but there was no significant difference between the groups (P > 0.05).</p><p><b>CONCLUSION</b>ICSI with epididymal or testicular sperm does not lead to more stillbirths or congenital malformations compared with ICSI using ejaculated sperm.</p>

Pregnancy outcomes of day-2 versus day-3 embryo transfer in poor responders / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate whether the day of embryo transfer (day 2 or day 3) affects clinical pregnancy outcomes in poor responder patients.</p><p><b>METHODS</b>We retrospectively analyzed the pregnancy rates of 265 initial fresh cycles of in vitro fertilization-embryo transfer (IVF-ET), all transferred on day 2 (n = 169) or day 3 (n = 96) irrespective of quality because of an extremely low number of available embryos.</p><p><b>RESULTS</b>Among the poor responders aged < 35 years, a higher rate of clinical pregnancy was achieved in the day-3 than in the day-2 group (50% vs 32.43% ; RR = 0.65, 95% CI: 0.43 - 0.99), and among those aged years, the two groups showed similar pregnancy outcomes.</p><p><b>CONCLUSION</b>Shortening the time of embryo culture has no obvious benefit for the pregnancy outcome. For the poor responders under 35 years of age, day-3 embryo transfer may afford an even higher rate of clinical pregnancy.</p>

Enhanced immune response of a novel T-cell immunogen in vaccine for foot-and-mouth disease / 生物工程学报

We investigated the enhanced immune response of a recombinant T cell immunogen as an effective cellular immune adjuvant. The T cell immunogen named TI contained several T cell epitopes from the VP1, VP4, 3A and 3D proteins of foot-and-mouth disease virus (FMDV) and two pan-T helper (T(H)) cell sites to broaden the immunogenicity of the protein. Meanwhile, another fusion protein named OA-VP1 was expressed in bacteria, which contained two VP1 proteins of O and Asia1 type FMDV. Mice were vaccinated with commercially inactivated vaccine or OA-VP1 protein with or without the TI immunogen. The results show that mice inoculated with inactivated vaccine or OA-VP1 protein supplemented with TI immunogen produced significantly higher level of neutralizing antibodies (P < 0.01 or P < 0.05) than the mice only inoculated with inactivated vaccine or OA-VP1 protein by microneutralization assay. An obvious increase in T cell number by flow cytometric analysis and significantly higher concentration of IFN-gamma secreted in culture media of spleen lymphocytes were observed in groups supplemented with TI immunogen (P < 0.01). TI immunogen was an effective stimulator for humoral and cellular immunity and could help improve the immunogenicity of inactivated vaccine or protein subunit vaccine.

Relationship between nasal inverted papilloma and human papillomavirus subtypes / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the distribution of human papillomavirus (HPV) subtypes in nasal inverted papilloma (NIP), and to evaluate the relationship between HPV and NIP.</p><p><b>METHODS</b>Twenty-one HPV subtypes were detected in paraffin-embedded tissues of 101 cases of NIP by flow through hybridization and gene chip (HybriMax), 24 cases of normal nasal mucosa were used as controls.</p><p><b>RESULTS</b>HPV positive rates of NIP were 64.36% (65/101). Benign NIP group, NIP with atypical hyperplasia group, NIP with cancerous group of HPV positive rates were 59.7% (46/77), 81.8% (18/22) and 50% (1/2) respectively. The control group was negative (0/24). The comparison between NIP group and control group was statistically significant (chi(2) = 32.178, P < 0.05). Benign NIP group and NIP with atypical hyperplasia group were compared, but no statistically significance (chi(2) = 3.649, P = 0.056) was found. The constituent ratio of benign NIP group and NIP with atypical hyperplasia group in high, low-risk HPV subtypes infections was compared, a statistically significance (chi(2) = 10.412, P < 0.05) was found.</p><p><b>CONCLUSIONS</b>The occurrence of NIP was related with HPV infection. High-risk HPV subtype infections or multiple infections will prompt benign NIP to NIP with atypical hyperplasia. Understanding the distribution of HPV subtypes in the NIP is helpful to predict the clinical behavior.</p>

Clinical application of oocyte vitrification in failed testicular sperm extraction cycles: report of 8 cases / 中华男科学杂志

<p><b>OBJECTIVE</b>To evaluate the clinical application value of oocyte vitrification in failed testicular sperm extraction cycles in non-obstructive azoospermia (NOA) patients.</p><p><b>METHODS</b>We retrospectively analyzed the clinical data of 8 women undergoing oocyte frozen-thawing cycles by vitrification because of failed testicular sperm extraction from their NOA husbands and no banked donor sperm on the day of oocyte retrieval. The oocytes were cryopreserved by vitrification with cryotop and thawed 2 months later. The surviving metaphase II (MII) oocytes were injected with the banked donor sperm of the same blood type as the husbands by intracytoplasmic sperm injection (ICSI) for fertilization. The rates of oocyte survival, fertilization, cleavage, good embryos and pregnancy were evaluated.</p><p><b>RESULTS</b>Sixty oocytes were vitrified and 47 (78.3%) survived after thawing, of which 41 MII oocytes underwent ICSI and 33 (80.5%) of them were fertilized. The rates of cleavage and good embryos were 81.8% (27/33) and 59.3% (16/27) respectively. Fifteen of the embryos were transferred to the 8 patients, with 1.9 +/- 0.8 per cycle, of which 5 (33.3%) were confirmed by ultrasound to have been implanted and 5 resulted in clinical pregnancy (62.5%), all singleton without miscarriage. Three normal boys and 1 normal girl were already born, with the pregnancy time of (39 + 4 +/- 0.4) wk and newborn body weight of (3787.5 +/- 513.7) g, respectively.</p><p><b>CONCLUSION</b>Vitrification of oocytes in failed testicular sperm extraction cycles is a promising technique for preserving female fertility, which, with ICSI of banked donor sperm, may result in satisfactory clinical outcomes.</p>

Outcomes of ICSI with microamount frozen-thawed sperm obtained by PESA or TESA in the treatment of azoospermia / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects and clinical pregnancy outcomes of intracytoplasmic sperm insemination (ICSI) with microamount frozen-thawed sperm obtained by percutaneous epididymal sperm aspiration (PESA) or testicular sperm aspiration (TESA) in azoospermia patients.</p><p><b>METHODS</b>We divided 365 azoospermia patients treated by ICSI into an experimental group (n = 123) and a control group (n = 242) , the former with microamount frozen-thawed sperm, and the latter fresh sperm obtained by PESA or TESA. The rates of fertilization, good embryos, clinical pregnancy, miscarriage, ectopic pregnancy and multiple pregnancy were analyzed and compared between the two groups.</p><p><b>RESULTS</b>With PESA, the experimental group showed no statistically significant differences from the control group in the rates of fertilization (75.67% vs 76.49%), good embryos (64.96% vs 66.09%), clinical pregnancy (55.21% vs 57.22%), clinical miscarriage (13.21% vs 12.61%), ectopic pregnancy (3. 77% vs 5.41%) and multiple pregnancy (37.74% vs 37.84%) (P > 0.05); nor with TESA (74.41% vs 76.43%, 64.63% vs 66.35%, 46.81% vs 53.39%, 18.18% vs 14.55%, 4.55% vs 1.82%, 37.74% vs 37.84%, P > 0.05). The revival rate of the frozen-thawed sperm from PESA was 70.07%, not significantly different from that of TESA (62.67%) (P > 0.05).</p><p><b>CONCLUSION</b>ICSI with frozen-thawed micro-amount sperm obtained by PESA or TESA is a safe, economic and effective method for the treatment of azoospermia. The techniques for reviving frozen sperm from PESA or TESA remain to be optimized, and whether these techniques may result in long-term genetic risks in the offspring deserves further investigation.</p>

Comparing the Oxygen Free Radicals and Oxygen Consumption of Leukocytes in Leukemia Patients and Healthy Persons / 中国实验血液学杂志

Using the electron spin resonance (ESR) and probe technique, the species of reactive oxygen free radicals and the oxygen consumption were observed in the metabolic process of oxygen of leukocytes from leukemia patients and healthy persons. Results showed that weak ESR spectrum of hydroxyl radical ((*)OH) signal was detected in the leukemia patient's leukocytes, there was no significant difference in oxygen consumption with or without PMA stimulation; superoxide anion (O(2)) in normal leukocytes was detected, and oxygen consumption increased markedly compared to the normal respiration without PMA stimulation (P < 0.001). There was no significant difference in oxygen consumption between leukemic and normal leukocytes without PMA stimulation; however, it was higher in normal leukocyte than in leukemic leukoeytes after PMA stimulation (P < 0.001). These observations suggest that dysfunction of respiratory burst is existed in leukemic leukocytes, and their oxygen consumption is markedly lower than that of normal leukocytes.