Objective To construct the prokaryotic expression vector containing XIAP and TAT gene.Methods XIAP gene was synthesized and the pTAT-HA plasmid was digested with NcoI and XhoI restriction enzyme,then rat XIAP gene was ligated by T4 DNA ligase with pTAT-HA plasmid to construct pTAT-XIAP expression plasmid encoding XIAP and TAT fusion protein.pTAT-XIAP vector was transformed into E.coli.DH5?.Transformants were cultured on LB medium and selected.The plasmids were extracted,purified and identified by agarose gel electrophoresis.Results A few of transformants with pTAT-XIAP on LB medium containing ampicillin were observed and a number of them on LB medium without ampicillin,none of untrasformed DH5? colonies were found on LB medium with ampicillin.The gel electrophoresis analysis showed that the molecular weight of recombinated pTAT-XIAP vector was 4500bp and the pTAT-HA vector was about 3000bp consistent with the atlas of the vector.Conclusion XIAP gene was cloned into pTAT-HA vector,pTAT-XIAP vector was successfully constructed.

In the process of promoting the implementation and transformation of the Q3D guideline of the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) in China, the risk assessment of elemental impurities in naturally sourced excipients often faces challenges such as multiple types of elemental impurities and insufficient detection methods. In this paper, an inductively coupled plasma mass spectrometry (ICP-MS) method for screening 12 elemental impurities in titanium dioxide was established using the optimized acid extraction pre-treatment method. The accuracy and repeatability of the method were good. The recoveries of cadmium, lead, mercury, cobalt, nickel, lithium, barium, molybdenum, copper, chromium and iron were 83.6%−101.6%, the recovery of antimony was 54.1%, and the RSD was 1.1%−10.2% (n=6). The evaluation results of elemental impurities showed that the risk of cadmium, mercury, cobalt, nickel, lithium, barium, molybdenum, copper, chromium and antimony was low, while the risk of lead was high, and it is recommended that a lead salt test be added to the pharmacopoeia standard for this product. Based on the statistical data, the differences in the control levels of elemental impurities and whiteness of 29 batches of titanium dioxide samples from 10 domestic and foreign manufacturers were compared and analyzed. The Pearson correlation coefficient method was used to analyze the correlation between residual elemental impurities and whiteness, and a heat map was used to visualize the results. The results showed that whiteness was significantly negatively correlated with the amount of residual barium and the sum of residual elemental impurities. It is suggested that the manufactures should include whiteness as an internally controlled indicator to further improve the quality of titanium dioxide used as a pharmaceutical excipient.

Objective To study the application of the array comparative genomic hybridization (Array-CGH) for the detection of chromosomal disorders in newborns.Method The Array-CGH technique was used to analyze the whole genome of the patients who were suspected of chromosomal disease in neonatal ward of our hospital from January to December in 2014,and further verification in genomic unbalanced ectopia was carried out by FISH (fluorescence in situ hybridization,FISH).Result Among 514 patients,104 were found carrying chromosomal abnormalities with a detection rate of 20.2％.The most common chromosomal disease is the Down syndrome syndrome (24 cases),followed by the chubby Willy and Angel syndrome(17 cases),while the Wolf-Hirschhorn syndrome in 5 cases,Williams syndrome in 5 cases and the Criduchat syndrone in 5 cases.The results of FISH were consistent with Array-CGH.Conclusion The technique of Array-CGH can be used to scan the whole genome of children with unknown disease.As a high-throughput and rapid research method,this technique has important clinical significance in the screening of chromosomal diseases.

Objective To compare complications incidence rates of 8-core and 12-core prostate biopsies guided with transrectal ultrasound retrospectively,and to study intervention ways to decrease complications.Methods The data of 260 consecutive patients undergoing first-time transrectal ultrasound-guided biopsy were analyzed.132 patients underwent 8-core biopsy and 128 patients underwent 12-core biopsy.Results In 8-core group,there were 7 cases of infections,15 cases of gross hematuria,7 cases of rectal bleeding and 6 cases of acute urinary retention,and in 12-core group,5 cases of infections,19 cases of gross hematuria,9 cases of rectal bleeding and 7 cases of acute urinary retention in 12-core group.There are no statistical differences of complications incidence rate between two groups (P ＞ 0.05).when prostate volume ＞ 45 ml,detection rate of prostate cancer in 12-core group was significantly higher than that in 8-core group.Conclusions There are no statistical differences of complications incidence rate between 8-and 12-core prostate biopsy.Homologous measure must be taken to reduce complications.12-core prostate biopsy is recommended as prostate volume ＞ 45 ml.

Objective To assess the efficiency and safety of mycophenolate mofetil (MMF) in patients with multiple sclerosis (MS) and neuromyelitis optica (NMO). Methods Twenty-seven patients with MS or NMO were selected, and the patients were divided into 2 groups:MMF group (MMF combined with glucocorticoid treatment group, 10 cases) and glucocorticoid group (only glucocorticoid treatment group, 17 cases). There were 5 cases with MS and 5 cases with NMO in MMF group. There were 13 cases with MS and 4 cases with NMO in glucocorticoid group. The therapeutic efficacy 6 months after treatment, expanded disability status scale (EDSS) before treatment and 6 months after treatment, and annualized relapse rate (ARR) were compared; and the safety was observed. Results There was no statistical difference in efficacy rate 6 months after treatment between MMF group and glucocorticoid group: 9/10 vs. 11/17, P>0.05. The EDSS scores 6 months after treatment in MMF group and glucocorticoid group were significantly lower than those before treatment: (2.41 ± 2.05) scores vs. (3.40 ± 2.05) scores and (1.17 ± 0.92) scores vs. (2.38 ± 1.28) scores, and there were statistical differences (P<0.05), particularly for the patients with MS. The ARR 6 months after treatment in MMF group was significantly lower than that before treatment: 0 time/year vs. 0.75 times/year, and there was statistical difference (P<0.05). The difference of ARR before and after treatment in MMF group was significantly higher than that in glucocorticoid group: 0.75 times/year vs.- 0.46 times/year, and there was statistical difference (P<0.01), particularly for the patients with MS. Only 1 female patient had myalgia when taking higher dosage of MMF, and the symptom tended to relieve after the dosage was reduced. Conclusions MMF is effective in the treatment of MS and NMO. MS can improve the neurological function and reduce the recurrence of the disease;and the safety is high.

Objective To discuss the value of magnetic resonance (MR) diffusion tensor imaging (DTI) technology in the diagnosis of the apparent diffusion coefficient (ADC) and the fraction anisotropy (FA) of neonates with hypoxic-ischemic encephalopathy (HIE) in the subacute stage.Methods Ten cases of normal full-term neonates and 34 term neonates diagnosed with HIE in the subacute stage were investigated with conventional MR and DTI.Of the 34 cases, 15 cases were mild HIE, 12 cases were moderate HIE,7 cases were severe HIE.ADC/FA values in different parts of the brain were analyzed.And the receiver operating characteristic (ROC) curve was used,the sensitivity and specificity of the FA in specific parts were also analyzed.Results (1) On convention MRI : according to the MRl scoring system,score of mild group was(2.6 ±-1.7) points, moderate group was (3.0 ± 1.5) points, severe group was (5.8 ± 1.3) points.There was significant difference between mild group and severe group, moderate and severe group (P ＜ 0.05).(2) The ADC and FA values in each group on DTI : ADC values in all of the regions of interest (ROIs) had no statistic difference between the HIE group and the controls(all P ＞ 0.05).FA values in posterior limb of internal capsule (PLIC) ,splenium had statistic difference between the HIE group and the controls.FA values in mild group were (0.317-0.018,0.339 ±0.025) ,moderate group (0.295 ±0.025,0.324 ±0.026) and severe group (0.252 ±0.039,0.253-0.036) and control group (0.455 ± 0.047,0.372 ± 0.032).(3) ROC analysis of the neonates diagnosed with HIE:the area under the ROC curve of FA values in PLIC, Splenium had statistic difference between mild group and moderate group (P ＜ 0.05).The area under the ROC curve in each group was 0.744 (P =0.032) ,0.658 (P =0.164).ROC curve was used to evaluate the diagnostic accuracy of FA values in PLIC between HIE group and control group,when FA value was lower than 0.337,0.803 was the largest area under the ROC curve, and there was significant difference between them(P ＜ 0.01).Conclusions FA values can reflect the brain white matter WM injury of HIE neonates quantitatively,and its diagnostic veracity is better than ADC value and conventional MR during the subacute phase of HIE.The diagnostic veracity of FA values in PLIC is higher than other parts.When the FA value is lower than 0.337, there is good sensitivity and specificity in diagnosis of HIE, so DTI technology can provide accurate and objective basis for clinical treatment with good sensitivity and specificity about the diagnosis of HIE in early stage.

Objective To understand the changes of schistosomiasis epidemic situation,so as to provide the evidence for for?mulating schistosomiasis control strategy in the Hexi reservoir area. Methods From 2012 to 2015,Xinyuan Village,Meishan Town in the north entrance of Hexi reservoir was selected as a monitoring site. According to the requirements of the monitoring program of schistosomiasis surveillance in Zhejiang Province,the Schistosoma japonicum infection was investigated by using the serological screening(IHA),and the basic situation of the surveillance site was also investigated. Results From 2012 to 2015,167 environments(21.68 hm2)were surveyed,and 2 slices(0.1 hm2)were found with Oncomelania hupensis snails. The detection rate of frames with snails was 0.12%,and the living snail density was 0.0192 snails per 0.1 m2. Totally 374 snails were dissected and no schistosome infected snails were found. A total of 970 local residents and 8 748 mobile people were investigated with the serological tests,and no schistosome infected people were found. In addition,3 085 cattle were investigated and no in?fected ones were found. Conclusion The schistosomiasis epidemic situation is stable in the Hexi reservoir area,but we still should strengthen the monitoring of imported source of infection and snail status,and increase the efforts of environmental trans?formation.

Aim Toestablishanexcellentratasthmamodel from using OVA+pertussis sensitized, OVA sensitized and per-tussissensitizedrats.Methods Thethreemethodswereusedto sensitize rats;methacholine bronchial provocation tests were per-formed to determine airway hyperresponsiveness;bronchoalveolar lavage fluid ( BALF) was prepared after the animals were chal-lenged by nebulized antigen. The differential white cell count in BALF was performed, and lung tissue was detected by morpho-logicalanalysis.Results AllofOVA+pertussissensitization,OVA sensitization and pertussis sensitization could deteriorate lung function, increase inflammatory cells and cause pathological change, and OVA + pertussis sensitized rat model had better effect than OVA sensitized and pertussis sensitized rat models. Conclusion OVA+pertussissensitizationandOVAaerosolisa successful rat asthma model.

objective To understand the status of Oncomelania hupensis snail distribution and diffusion in main drainages of Hexi Reservoir and evaluate the snail control effect of the schistosomiasis control engineering of Hexi Reservoir. Methods The O. hupensis snails were investigated by using the straw curtain method and fishing net method in different areas of the main drainages of Hexi Reservoir and the results were analyzed. Results A total of 1 800 straw curtains were used and 37 snails were found in Naxi stream. Totally 5 870 kg floats were salved and no snails were found. Conclusion The schistosomiasis con?trol engineering of Hexi Reservoir is effective in the prevention of the snail diffusion but there are still snails in the upstream. Therefore the snail surveillance and control need to be strengthened.

Objective:To analyze the variation and evolution characteristics of hemagglutinin (HA) and neuraminidase (NA) genes of influenza B virus circulating in Jining from 2017 to 2020.Methods:Throat swab specimens were collected from patients with influenza-like symptoms in sentinel hospitals and influenza outbreaks in Jining from 2017 to 2020 and tested for influenza B virus nucleic acid. After virus isolation, 20 representative strains of influenza B virus were selected to sequence the full length of HA and NA genes. Phylogenetic trees were constructed and the molecular characteristics were analyzed using bioinformatics software. Results:A total of 4 575 specimens were collected and 842 of them were positive for influenza virus, including 398 (8.7%, 398/4 575) influenza B virus-positive specimens. The positive rate of influenza B virus was 47.27% (398/842). The isolated influenza B virus strains of Victoria (BV) and Yamagata (BY) lineages from 2017 to 2020 shared 98.7%-98.8% and 98.5%-99.1% homology in HA gene with vaccine strains, respectively. The BV lineage strains isolated from 2018 to 2020 belonged to Victoria clade 1A branch and the BY lineage strains isolated from 2017 to 2018 belonged to Yamagata clade 3 branch. Mutations were detected in several antigenic sites, but not in the sites related to NA inhibitor resistance. Conclusions:Mutations in several antigenic sites caused antigenic changes in influenza B virus of BV and BY lineages, which might be related to the outbreaks of influenza B virus infection in Jining during 2017 to 2020.