Objective To clone Tp0821,Tp0319,Tp0971 and Tp0663 gene,construct prokaryotic expression plasmid,the ex-pression of recombinant proteins,purification and used to detect syphilis patients serum antibodies.To explore Tp0821, Tp0971,Tp0319 and Tp0663 recombinant protein in the diagnosis of Tp infection,rich library of candidate antigens syphilis serology diagnosis.Methods From Tp library screening Tp0821,Tp0319,Tp0971 and Tp0663 four kinds of membrane pro-tein,through the analysis of the bioinformatics software selection advantage antigen epitope,connected with pQE32 carrier respectively built pQE32/Tp0821,pQE32/Tp0319,pQE32/Tp0971 and pQE32/Tp0663 prokaryotic expression recombi-nant;In vitro cloning,expression and purification and on single Tp0821,Tp0319,Tp0971,Tp0663 and four kinds of recombi-nant protein chimeric antigen (Tp0821-Tp0319-Tp0971-Tp0663)in order to establish the indirect Tp-envelope antigen ELISA method,based on TRUST and TPPA method comparison,detection collected from January 2013 to June 2014 in Shenzhen Baoan District People’s Hospital outpatient and hospitalization Tp negative patients,160 cases of positive speci-mens of 83 cases of clinical and Tp,and evaluated its application in the syphilis serology diagnosis.Results Successful con-structed of prokaryotic expression vector pQE32/Tp0821,pQE32/Tp0319,pQE32/Tp0971 and pQE32/Tp0663.Efficient expressed and purified their relative molecular mass of recombinant proteins.Seted up Tp-indirect ELISA method to detect 160 cases of Tp negativepositive clinical specimens and 83 cases of Tp,and compared with TPPA,the sensitivity were 85.5% (71/83),84.3% (70/83),89.2% (74/83),81.9% (68/83)and 95.2% (79/83)respectively,specificity was 100%(160/160),and the coincidence rate was 82.6%~95.7%.Single Tp0821,Tp0319,Tp0971 and Tp0663 recombinant protein positive rate of TP-ELISA was established (85.5%,84.3%,89.2% and 81.9%)compared with TPPA method of positive detection rate (100%)had significant difference statistically significant (χ2= 24.5~31.8,P<0.01),and four kinds of re-combinant protein chimeric antigen positive rate of TP-ELISA was established with the TPPA method the positive detection rate of comparative difference was statistically significant (χ2=7.95,P<0.05).Conclusion Preparation Tp0821,Tp0319, Tp0971 and Tp0663 recombinant protein had good immune activity,for the further study of its application in the diagnosis of syphilis serology lay a certain foundation,but in four kinds of recombinant protein chimeric antigen based Tp Tp-indirect ELISA method for serological diagnosis,can improve the detection sensitivity.

The absorption-elution test using low ionic strength solution (LISS) has been compared with the test using normal saline in MN typing of 258 bloodstain samples stored 1 to 6 years. The accuracy rate was 94.57％ using LISS method. The present study indicated that the LISS method is more sensitive than tests carried out in normal saline.

Three SNaPshot multiplex assays were developed to test 23 coding region single nucleotide polymorphisms (SNPs) and one control region SNP outside hypervariable regions (HVR)I and II, which was aimed at increasing the discrimination power of the mitochondrial DNA (mtDNA) typing in forensic casework, and confirming haplogroup assignments of mtDNA profiles in both human population studies and medical research. The selected SNPs targeted the East Asian phylogeny. These multiplex assays were validated by comparing with the sequencing analysis of samples chosen randomly. The mtDNA variations of 100 unrelated individuals from the Wuhan population in China were examined and classified into 31 haplotypes, and the haplotype diversity was estimated to be 0.952. The multiplex SNaPshot method is rapid and robust, and suitable for large-scale screening studies of mtDNA variability.

Objective To study the effects of asiaticoside on the growth of melanoma B16 cell cultures in vitro. Methods Melanoma B16 cells were subcultured and the inhibition of cellular growth was investigated. The morphology of the cells was observed after inhibition. The induction of apoptosis by asiaticoside was determined by flow cytometry. Results It was found that asiaticoside could significantly inhibit the growth of B16 cell cultures in vitro in a dose-dependent manner. The annexin-v positive cells were increased, along with that cells intaking R123 marked mitochondria were decreased, and PI positive cells increased, which indicated that cellular apoptosis was induced. Conclusion Asiaticoside plays an inhibitory role in the growth of melanoma B16 cells.

BACKGROUND: The degradation speed of biological materials is critical for the clinical use of guided bone regeneration.The partial biological characteristics and treatment efficacy of concentrate growth factor (CGF) fibrin have been explored preliminarily, but its degradation properties have not yet been reported.OBJECTIVE: To explore the degradation properties of CGF fibrin and platelet-rich fibrin (PRF) in artificial saliva and compare the degradation speed of these two biological products.METHODS: Ten volunteers were selected, and 18 mL of venous blood from each volunteer was extracted and stored in two vacuum blood collectors. The blood samples were then placed into the drum of the Medifuge centrifugal acceleration machine, to separate CGF fibrin and PRF specimens following the preparation process, respectively. Both CGF fibrin and PRF specimens were respectively made into bulk and membranoid, and were then immersed in artificial saliva under 37 ℃. The mass of the bulk specimens and area of the membranoid specimens were measured regularly, and the degradation processes of CGF and PRF were recorded. The degradation curves were drawn to compare the degradation speed of CGF fibrin and PRF.RESULTS AND CONCLUSION: The mass of CGF fibrin and PRF showed no significant difference at the 5th day (P >0.05), while the mass of CGF fibrin was higher than that of PRF at the 3rd, 4th and 6th days (P < 0.05). The residual area of CRF was significantly larger than that of PRF at posttreatment 1-6 days (P < 0.05). To conclude, the degradation speed of bulk or membranoid CRF is slow than that of PRF in artificial saliva. The higher the fibrin content is, the slower the degradation ability is, indicating the strong bioreproductive function.

Objective:To explore the effect of progressive core strength training on balance of the old adults. Methods:From April to September, 2018, 80 old persons were divided into control group (n = 40) and intervention group (n = 40), who accepted physical exercise and core strength training, respectively, for twelve weeks. They were assessed with one-legged standing time with eyes closed, center of pressure (COP) track as standing with eyes closed in 30 seconds, maximum distance of COP tilting, Timed "Up and Go" Test (TUGT) and standing up and sitting down in 30 seconds. Results:The one-legged standing time with eyes closed, COP track as standing with eyes closed in 30 seconds, maximum distance of COP tilting, TUGT and standing up and sitting down in 30 seconds improved after training (t > 6.446, P < 0.001), and they improved more in the intervention group than in the control group (t > 2.127, P < 0.05). Conclusion:Progressive core strength training can improve the dynamic and static balance, body control and core strength for the old adults, which may prevent them from falling down.

Objective To investigate the risk factors for cognitive impairment and their correlation with collateral circulation in patients with acute ischemic stroke.Methods From January 2016 to December 2017,patients with acute ischemic stroke admitted to the Department of Neurology,Jiangmen People's Hospital were enrolled.According to their status of collateral circulation,they were divided into either good collateral circulation group or poor collateral circulation group.According to the Montreal cognitive score,they were divided into cognitive impairment group and non-cognitive impairment group.Multivariate logistic regression analysis identified the independent risk factors for cognitive impairment in patients with acute ischemic stroke.Results A total of 225 patients were enrolled,including 106 females (47.1％) and 119 males (52.9％),aged 47-87 years,average 66.96 ± 9.65 years.The average baseline score of the National Institutes of Health Stroke Scale was 4.5 ± 2.1.Ninety-seven patients (43.1％) had cognitive impairment;79 had obvious stenosis or occlusion at the intracranial and extracranial segments of cerebral arteries;148 (65.8％) had good collateral circulation,and 77 (34.2％) had poor collateral circulation.The proportions of patients with atherosclerotic stroke (P =0.033) and cognitive impairment (P =0.041) in the good collateral circulation group were significantly higher than those in the poor collateral circulation group.There were significant differences in the proportions of patients with large-artery atherosclerotic stroke (P =0.007),anterior circulation stroke (P=0.018),collateral dysfunction (P=0.041),as well as baseline systolic blood pressure (P =0.006),baseline diastolic blood pressure (P =0.013),low-density lipoprotein cholesterol (P=0.021),and homocysteine (P=0.024) between the cognitive impairment group and the non-cognitive impairment group.Multivariate logistic regression analysis showed that baseline systolic blood pressure (odds ratio[OR] 1.007,95％ confidence interval[CI] 1.001-1.012;P =0.036),and large-artery atherosclerotic stroke (OR 2.650,95％ CI 1.490-4.714;P =0.001) were independently positively correlated with cognitive impairment,while the status of collateral circulation was not an independent risk factor for cognitive impairment (P =0.073).Conclusions The status of collateral circulation was associated with cognitive impairment in patients with acute ischemic stroke,but it was not an independent risk factor.

Objective: To explore the main characteristics of syndromes in traditional Chinese medicine (TCM) in post-stroke depression (PSD) and to provide basis for treatments with TCM herbs. Methods: According to diagnostic criteria of PSD, stroke patients and depression patients from Department of Neurology, First Affiliated Hospital, Anhui University of Traditional Chinese Medicine were assigned into cerebral stroke group (150 cases), depression group (151 cases) and PSD group (123 cases). Neuropsychological assessments and imaging and biochemical analyses were conducted. TCM syndrome differentiation for these diseases was performed. We also determined the characteristics of TCM syndromes of PSD, relative risk of the syndromes and their correlations with ages as well. Results: Scores of qi stagnation and blood stasis, liver qi depression, and transformation of fire due to qi stagnation in PSD group were significant higher than those in cerebral stroke group (P<0.05, P<0.01). In cerebral stroke group, majority of the patients displayed one syndrome, while in PSD and depression groups, the patients had three or more syndromes. Of these syndromes, the incidence rate of syndrome of liver qi depression complicated with transformation of fire due to qi stagnation or flaring of fire due to yin deficiency was high. The syndrome of liver qi depression occurred much more frequently in PSD group and depression group than in cerebral stroke group (P<0.05, P<0.01). The logistic regression analysis showed that the syndrome of qi stagnation and blood stasis had high relative risk to PSD. The syndrome of deficiency of heart and spleen was positively correlated with age in cerebral stroke group. Conclusion: The main TCM syndromes of PSD and depression are qi stagnation and blood stasis, liver qi depression, and transformation of fire due to qi stagnation. The syndrome of deficiency of heart and spleen is closely related to age among the stroke patients. The syndrome of qi stagnation and blood stasis serves as an independent risk factor for PSD. The more complicated the syndromes are, the more serious depression becomes.

Three SNaPshot multiplex assays were developed to test 23 coding region single nucleo-tide polymorphisms (SNPs) and one control region SNP outside hypervariable regions (HVR) Ⅰ and Ⅱ, which was aimed at increasing the discrimination power of the mitochondrial DNA (mtDNA) typing in forensic casework, and confirming haplogroup assignments of mtDNA profiles in both hu-man population studies and medical research. The selected SNPs targeted the East Asian phylogeny. These multiplex assays were validated by comparing with the sequencing analysis of samples chosen randomly. The mtDNA variations of 100 unrelated individuals from the Wuhan population in China were examined and classified into 3 i haplotypes, and the haplotype diversity was estimated to be 0.952. The multiplex SNaPshot method is rapid and robust, and suitable for large-scale screening studies of mtDNA variability.

Objective To express and purify GRIK3 intracellular region protein in prokaryotic cell system. Methods The histamine(His)tagged GRIK3 intracellular region recombinant plasmid pCzn1-GRIK3-in-tra was constructed and transformed into Escherichia coli(E.coli). The His-GRIK3-intra recombinant protein was expressed after the induction with IPTG. The target protein was purified by Ni-NTA affinity chromatography column. Results The prokaryotic expression plasmid pCzn1-GRIK3- intra was successfully constructed and the GRIK3 intracellular region protein in E.coli were efficiently expressed after the induction with IPTG. The purified target pro-tein was successfully obtained by Ni-NTA affinity chromatography column. Conclusion The successful construc-tion of prokaryotic expression plasmid expressing GRIK3 intracellular region gene and preparation of high purity GRIK3 intracellular region protein have paved the way for further exploration of the intracellular signal transduction mechanism of membrane protein GRIK3.