Objective To construct a recombinant plasmid DNA containing herpes simplex virus type 1(HSV-1) glycoprotein D (gD) gene.Methods The HSV-1 gD gene was obtained by polymerase chain reaction (PCR) and inserted into TA cloning vector pGEM-T, then cloned into the eukaryotic expression vector pcDNA3.1 to generate pLy-D. The recombinant plasmid pLy-D, which was confirmed by partial sequencing and restriction endonuclease analysis, was transfected into Cos-7 cells and used to inoculate ICR mice via muscular injection. Immunohistochemistry and enzyme-linked immunoabsorbent assay (ELISA) were employed to test the gD expression in transfected cells and the specific anti-HSV-1 antibody in the serum of immunized mice, respectively.Results The gD eukaryotic expression plasmid pLy-D was constructed. Using the immunohistochemistry technique, the gD expression in pLy-D-transfected cells was detected. The ELISA demonstrated that specific anti-HSV-1 antibody could be induced in immunized mice after three times injection.Conclusions We constructed HSV-1 gD eukaryotic expression plasmid pLy-D which could express gD protein in transfected cells and could induce humoral immune response in mice. This observation will be helpful in designing HSV prophylactic vaccine.

Objective To investigate the relationship between rapists and related allele genes based on the analysis of 15 short tandem repeats (STRs) loci genetic polymorphism. Methods The method of Genome-wide scan was being used. Buccal swab samples of 129 rapists and 156 random populations were collected and PCR compound amplification was carried out with the aid of AmpFISTR Identifiler system. Then the products were subjected to electrophoresis and gene detection with AB13100 type gene analysis system so as to calculate and compare the alleles of 15 STRs gene frequency in the two groups. Results All the 15 STRs loci allele gene frequency in rapists and random population was found to coincide with Hardy-Weinberg law(P>0. 05). Allele 28 of D21S11 (rapists: 1.55% ,control group:5. 13%) ,allele22 of FGA(rapists:24.03% ,control group:16.99%),allele23 of FGA(rapists: 17.05% ,control group:26.28%) ,allele 10 of TH01(rapists:1.16% ,control group:4.17%) ,allele 8 of TPOX(rapists:55.77% ,control group:63.77%),allele 12 of TPOX(rapists:4.26% ,control group: 1.28%) were different between the two groups (P< 0.05) .while it is no differ significantly in other STRs loci allele gene(P >0.05). Conclusion Allele 28 of D21 S11,allele 22 and 23 of FGA, allele 10 of TH01, allele 8 and 12 of TPOX may be associated with the violent crime of rape. It is suggested that there are existing sensitive or resistance genes about the violent crime of rape in chromosome 2,4,11,21.

We analyzed genetic evolution characteristics of avian influenza A (H7N9) virus isolated in Zhaoqing,China,2014-2016.Nucleic acid were extracted and sequenced from 17 samples of H7N9 positive cases in Zhaoqing.Genetic characteristics of homology and important amino acid sites were analyzed by using BioEdit5.0 and MEGA6.0.The evolutionary trees were constructed by Neighbor-Joining and the referenced sequences were downloaded from GenBank,Eight nucleic acid fragments from 7 strains of H7N9 viruses were successfully generated.The highest homology was found in HA gene with A/chicken/Dongguan/695/2014(H7N9),and NA gene with A/chicken/Dongguan/1075/2014(H7N9).The internal genes were high homology with avian H7N9 and H9N2 virus from Dongguan and Shenzhen in Guangdong,China.The HA and NA genes were directly evolved in the Pearl River Delta evolution branch with the H7N9 sequences from the cities of Dongguan,Guangzhou and Shenzhen,while the sequences from the provinces of Anhui,Zhejiang,and Jiangsu were in the Yangtze River Delta evolution branch.There were 2 alkaline amino acids in cleavage site of HA,2 mutations (G186V and Q226L) in the crucial sites related with the receptor of HA protein,1 mutation (E627K) in PB2 protein,and 1 drug resistance mutation (S31N) in M2 protein.And no evidence of neuraminidase resistance in NA protein was found.In conclusion,the H7N9 virus for human infection in Zhaoqing may originate from avian H7N9 and H9N2 viruses,which circulated in the Pearl River Delta region of Guangdong from 2013 to 2014.The mutations of G186V,Q226L and E627 K might be related with high susceptibility to human beings.

Objective To study the effects of low calcium dialysate on bone mineral density (BMD) in old patients with low turnover renal osteodystrophy under maintenance hemodialysis.Methods Totally 72 elderly patients aged≥ 60 years under MHD for 6 months or more with parathyroid hormone(iPTH)＜100 ng/L were selected and randomly divided into treatment group(n=36,calcium 1.25 mmol/L in dialysate)and a control group(n =36,calcium 1.5 mmol/L in dialysate),for 12 months of treatment.The changes of albumin-corrected calcium and phosphorus,calciumphosphorus product,iPTH level,bone mineral density,and other indicators as well as related adverse events were observed before and 12 months after the study.Results There was no statistically significant difference in general conditions and the correlated laboratory examinations between the two groups before and after treatment (P ＞ 0.05).After dialytic treatments with dialysate containing calcium 1.25 mmol/L for 12 months,the therapy group versus pre-therapy and control group showed statistically significant decrease in parameters of mean arterial pressure(MAP) [(88.6 ± 9.2) vs.(92.6±10.4)and(93.7±8.8)mmHg],serum calcium[(2.4±0.1)vs.(2.6±0.3)and(2.6±0.2)mmol/L,t =5.061,5.074],phosphorus[(2.0±0.2)vs.(2.1 ±0.2)and(2.1±0.3)mmol/L,t=2.276,2.271],calcium-phosphorus product[(4.7 ± 0.5) vs.(5.3 ± 0.6) and (5.4 ± 0.7) mmol2 / L2,t =4.682,4.627](all P＜0.05),and showed statistically significant increase in parameters of iPTH levels[(132.6 ±37.8) vs.(71.3 ± 11.48) and (69.7 ± 16.0) ng/L;t value 8.824 and 9.048,respectively],bone mineral density values(Lumbar:0.8±0.9 vs.-1.2±1.1 and-1.2±1.1;t value 2.170 and 2.170,respectivly.Femoral neck:-0.8± 1.0 vs.1.3±1.2 and-1.3±1.3;t value 2.258 and 2.243,respectively) (all P＜0.05).In the control group after 12 months of treatment with calcium 1.5mmol/L dialysate,there was no significant difference in related parameters (P＞ 0.05).There was no significant difference in the adverse reactions between study and control groups (P ＞ 0.05).Conclusions The low calcium dialysate(calcium 1.25 mmol/L)used in elderly patients with iPTH＜100 ng/L under MHD can effectively improve the excessive depression of parathyroid function and the decreased BMD,and better control the albumin corrected calcium,phosphorus,calcium-phosphorus product level,and has a good security.

Objective: To investigate the willingness to receive measles-containing vaccine (MCV) and its influencing factors among healthcare workers in the Yangtze River Delta region, so as to provide the evidence for improving the measles-containing vaccination rate Methods: Healthcare workers were sampled from 19 medical institutions in each of Shanghai Municipality, Jiangsu Province, Zhejiang Province and Anhui Province for questionnaire surveys using a multi-stage stratified convenience sampling methods from July 2020 to March 2021. Participants' gender, age, educational level, professional title, measles-containing vaccination, awareness of MCV and willingness to receive MCV were collected, and the factors affecting the willingness to receive MCV were identified among healthcare workers using a multivariable logistic regression model. Results: Totally 1 403 questionnaires were allocated, and 1 394 valid questionnaires were recovered, with an effective recovery rate of 99.36%. The respondents included 327 men and 1 067 women, with a male to female ratio of 1∶3.26, and 64.35% (897) were at ages of 31 to 50 years. There were 1 005 respondents with a bachelor degree (72.09%), 765 with middle and senior professional titles (54.88%), 676 with a history of measles-containing vaccination (48.49%), 1 176 with willingness to receive MCV (84.36%) and 218 without willingness to receive MCV due to convenience of vaccination (30.73%) and cost (19.27%). Multivariable logistic regression analysis showed that region (Zhejiang, OR=1.613, 95%CI: 1.054-2.470; Anhui, OR=2.058, 95%CI: 1.259-3.363), and no history of measles (OR=2.219, 95%CI: 1.302-3.781) were factors improving the willingness to receive MCV among healthcare workers, and hospital level (secondary, OR=0.483, 95%CI: 0.306-0.763; tertiary, OR=0.251, 95%CI: 0.160-0.394), history of measles-containing vaccination (no, OR=0.262, 95%CI: 0.172-0.399; unknown, OR=0.386, 95%CI: 0.266-0.559), and unawareness of MCV knowledge (OR=0.208, 95%CI: 0.081-0.536) were factors inhibiting the willingness to receive MCV among healthcare workers. Conclusions The willingness to receive MCV correlates with region, history of measles, hospital level, history of measles-containing vaccination and awareness of MCV knowledge among healthcare workers in the Yangtze River Delta region.

Objective: To establish a TaqMan-MGB probe-based real-time fluorescence RT-PCR assay for avian influenza H5N6 virus used in rapid diagnosis for suspected cases and surveillance for outer environment of live poultry markets. Methods: Based on the conservative sequences of avian influenza H5N6 virus for HA and NA gene published on GenBank, specific primers and TaqMan-MGB probes were designed to develop and optimize for the dual real-time RT-PCR assay. Specificity, sensitivity, repeatability and comparison tests were carried out. Results: This dual real-time RT-PCR detection can be completed within 80 minutes. There was no cross-reaction with other subtypes of influenza virus and common respiratory pathogens. The minimum detection limit could be up to 10 copies/reaction. The correlation coefficient of standard curve for the gene of H5 and N6 were 0.999 and 0.993, and the coefficients of variation for cycle threshold were range from 0.151%-0.549%and 0.213%-0.575%, respectively. The positive and negative coincidence rates of the validation test were 100%. Conclusions This TaqMan-MGB probe-based dual real-time RT-PCR for avian influenza H5N6 virus was rapid, specific and sensitive. It will have a good use in early emergency detection of suspected cases and continuous monitoring of external environment in live poultry trade market.

OBJECTIVE Disturbance of K+ ion balance in inner ear is associated in age-related hearing loss. Our study is to investigate the role of NKCC1 and Na-K-ATPase in cochlea and auditory function regulated by with different expression of NKCC1 and Na-K-ATPase. METHODS Auditory threshold of young or old C57BL/6J mice was measured by auditory brainstem response(ABR). The expression of NKCC1 and Na-K-ATPase in mice cochlea were evaluated by reverse transcription polymerase chain reaction(RT-PCR) and western blotting. Furosemide and Ouabain were applied in vivo to inhibit NKCC1 and Na-K-ATPase in C57BL/6J mice. RESULTS C57BL/6J mice developed hearing loss at 12M by ABR threshold shifting to (75±10), (78±26) and (81±14)dB SPL at frequencies of 8, 16 and 32 kHz; PCR showed that the relative expression of NKCC1 and Na-K-ATPase mRNA in the aged group decreased, which were 0.52±0.06 and 0.35±0.04 times higher than those in the young control group, the difference was statistically significant(t =7.466 and 16.11, all P＜0.05). WB showed that relative expression of NKCC1 and Na-K-ATPase protein level in the aged group decreased by 0.79±0.02 and 0.68±0.05 times as much as that of the young control group, the difference was statistically significant(t =8.857 and 6.771, P all＜0.05). After applied with Furosemide and Ouabain to suppress the two ion transporters, the ABR threshold increased to (50±17), (53±21), (55±17)dB SPL and (56±6), (70±17), (73±6)dB SPL at frequencies of 8, 16 and 32 kHz. CONCLUSION In vivo experiment of C57BL/6J suggested that NKCC1 and Na-K-ATPase might be related to age related hearing loss.

Objective : To analyze the value of virtual surgical planning in the surgical treatment of osteoradionecrosis of the mandible and to provide a reference for clinical practice. .Methods : From September 2017 to June 2018, 13 patients with mandibular osteoradionecrosis were evaluated preoperatively using the 3D virtual surgery software CMF Proplan 2.0. The surgical guide was designed and 3D printed. Bone resection, fibula shaping and bone graft localization were completed during the operation. In some cases, implants were implanted at the same time, and denture restoration was completed 3 to 6 months after surgery. Patients’ general information, perioperative data, and efficacy evaluation were analyzed. Results: All patients underwent surgery successfully. The survival rate of the free fibula musculocutaneous flap was 100% (13/13), and one patient had complications (partial necrosis at the edge of the flap). The follow-up period was 7 to 15 months, and the median time was 10 months. All patients achieved a healing effect. The number of cases with an increase in mouth opening ≥ 1 cm, 0.5 cm ≤ mouth opening increase < 1 cm, and mouth opening increase < 0.5 cm were 5, 6, and 2, respectively. An imaging examination showed that 12 patients had good bone healing, and 1 patient did not completely heal 7 months after operation. The denture restoration was 92.3% (12/13), of which 3 cases were implanted and repaired at the same time. The average chewing efficiency was 56.11% ± 7.12% (42.03%-67.83%). Conclusion Virtual surgical planning is an effective method for the surgical treatment of mandibular osteoradionecrosis, which can reduce the risk of surgery and more effectively perform mandibular shape and function repair.