Objective To discuss the effect of DNA extraction of bloodstain on the filter paper with four methods of solid phase absorption.Methods 180 bloodstain samples on the iflter paper, each one contains 1 microlitre anticoagulation peripheral venous blood, divided into 4 groups with 45 samples, respectively. All samples were treated with four methods of solid phase absorption, i.e. DNA IQ? System, D-shield sensitive DNA Extraction Kit, High efficiency Silica Bead DNA Extraction Kit and Conventional silica bead method. The concentration of DNA and the results of STR typing of four groups were compared each other.Results The concentration of DNA was 3.764±1.790μg/mL and 3.634±1.112μg/mL by using D-shield sensitive DNA Extraction Kit and High efifciency Silica Bead DNA Extraction Kit, respectively. However, the concentration of DNA by using Conventional silica bead method group (3.350±1.250) was not signiifcantly different from each other (P<0.05), while the concentration of DNA extracted with above three methods were higher than by using DNA IQ? System (1.864±1.207)(P<0.001); Signiifcant differences of peak height existed between DNA IQ? System and other three methods (P<0.001); As the same time, the peak height of samples by using High efficiency Silica Bead DNA Extraction Kit and Conventional silica bead method were signiifcantly different from D-shield sensitive DNA Extraction Kit (P<0.01).Conclusions The DNA extracted in bloodstain on the iflter paper by using D-shield sensitive DNA Extraction Kit, High efifciency Silica Bead DNA Extraction Kit and Conventional silica bead method was more than DNA IQ? System. Meanwhile, the quality of DNA using High efifciency Silica Bead DNA Extraction Kit and Conventional silica bead method may be higher.

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.

Objective To identify the feasibility and the anatomical parameters of posterior transarticular pedicle screw fixation in the thoracic spine, provide a reference for clinical applications. Methods Twenty human cadaveric thoracic spine segments were dissected posteriorly and anteriorly, with care taken to expose the laminas and pedicles. The entrance point of transarticular pedicle screws was located in the 7 mm away from the above at the inferior margin of the lamina and the inside at the exterior margin, respectively. Posterior transarticular pedicle screws implantation was performed under direct visualization into T1,2,T5,6 and T9,10 Under direct abservation, the feasibility of posterior transarticular pedicle screw fixation was assessed. Then a CT was done. On the morphologic CT scan, the angle and length of the transarticular pedicle crew trajectory were measured. Results The thoracic transarticular pedicle screw trajectory were caudal tilting in the sagittal plane and lateral tilting in the axial plane with successful placement. Screws were placed across the facet joint, and from the inferior articular process of upper thoracic vertebra into the pedicle of lower thoracic vertebra. There was little difference between different number of thoracic vertebrae of the angle, but without significance. The average angles of the screws were 41.4°+3.2°caudal tilting in the sagittal plane and 2.1°±0.7° lateral tilting in the axial plane. The average trajectory lengths were (40.6±4.9)mm, and the lengths increased gradually from upper thoracic vertebra to middle and lower. There were significant differences statistically among T1,2, T5,6 and T9.10 (F=74.09, P＜0.01 ). Conclusion Posterior transarticular pedicle screw fixation is feasible, and there are some directions for implantating the screws. Transarticular pedicle fixation in the thoracic spine affords an alternative to standard pedicle screw placement for thoracic stabilization.

Objective To establish the enzyme-linked immunosorbent assay (ELISA) methods for the quantitative determination of IgG antibodies against diphtheria (DT) and tetanus (TT).MethodsPurified diphtheria toxiod and tetanus toxoid were respectively used as the coating antigens,the human-derived serum antibody standard substance of DT and TT served as the standard substance.The dose-response curves of the tested samples and standard substance were fitted.Then the two quantitative ELISA methods for determining the antibody to DT (Anti-DT) and antibody to TT (Anti-TT) were established with the parallel lines method.Then the methodological verification and application study were conducted.Results The validation results of the two quantitative ELISA measurement methods were in accordance with the regulations.The quantity limit of ELISA method for quantitative detection of Anti-DT demonstrated to be 0.084 mIU/mL,its average recovery rate was 97.6%.The intra-assay coefficient of variation(CV) and inter-assay CV of this Anti-DT assay were ≤ 3.40% and ≤5.05%,respectively.The quantity limit of ELISA method for quantitative detection of Anti-TT demonstrated to be 0.175 mIU/mL,its average recovery rate was 97.5%.The intra-assay CV and inter-assay CV of this Anti-TT assay were ≤ 2.42% and ≤5.58%,respectively.These two methods were applied for the immunogenicity evaluation after infantile basic immunization by diphtheria and tetanus vaccines.Conclusion The two established quantitative ELISA methods demonstrate high accuracy and good reproducibility,which are suitable for the ordinary laboratory to carry out the work and can be used in the serological effect evaluation after diphtheria and tetanus vaccine immunization and epidemiological study of diphtheria and tetanus disease.

Objective To compare the PCR effect of rapid PCR instrument and common PCR instrument. Methods The concentration of 9947A standard was diluted to 0.1, 0.05, 0.025, 0.0125, 0.0063 ng/μL, 100 samples from each concentration group to establish PCR reaction system with Identifiler? Plus PCR Amplification Kit, 50 samples of them test with rapid PCR instrument (Speed cycler2 thermocycler), the other 50 samples test with common PCR instrument (9700 thermocycler). Detection of PCR product with 3500xL Genetic Analyser, the STR typing of both groups of each concentration group should be compared. Results The success rate of both thermocyclers have no significant difference (P>0.05); The success number of STR typing of common PCR instrument (13.7±1.0; 11.3±1.5) were higher than rapid PCR instrument (13.1±1.3; 9.9±1.9) when the concentration of 9947A were 0.0125, 0.0063ng/μL (P=0.029; P<0.001); The peak height of DNA typing map obtained from common PCR instrument (18931±4625;13437±3165; 5752±1344) were higher than rapid PCR instrument (16929±4034; 11815±4120; 4865±1401) when the concentration of 9947A were 0.1, 0.05, 0.025ng/μL (P=0.023; P=0.030; P=0.002). Conclusions The rapid PCR instrument could achieve the equal success rate to the common PCR instrument with less time, which revealed that the rapid PCR instrument was suitable for application in practical cases; The quality of STR typing from common PCR instrument may be more higher.

Objective:To explore the effect of traditional Chinese medicine fumigation and washing on the clinical indicators of elderly patients with distal radius fractures after small splint fixation.Methods:From November 2015 to November 2018, 180 elderly patients with fracture of the distal radius in Ningbo Hospital of Traditional Chinese Medicine were selected, and they were divided into control group and observation group according to the random digital table method, with 90 cases in each group.The two groups were given manual reduction and small splint fixation treatment, and the observation group was treated with traditional Chinese medicine fumigation on the basis of the control group therapy.The related inflammatory cytokines in patients plasma and serum viscosity(TNF-α, IL-1, IL-6) were detected, and the clinical curative effect and adverse reactions were observed.Results:Before treatment, there were no statistically significant differences in plasma viscosity and blood inflammatory factors(TNF-α, IL-1, IL-6) between the two groups( t=-0.171, -0.126, 0.483, 0.459, all P>0.05). After treatment, the above indicators in the two groups were significantly lower than those before treatment(observation group: t=38.970, 14.684, 72.258, 53.612; control group: t=20.098, 19.045, 33.962, 34.822, all P<0.05), which in the observation group were lower than those in the control group ( t=-30.239, -7.043, -31.815, -10.359, all P<0.05). The total effective rate of the observation group was 88.88%(80/90), which was higher than 71.11%(64/90) of the control group, the difference was statistically significant between the two groups(χ2=8.889, P<0.05). Conclusion:For the elderly patients with distal radius fracture, the combination of reduction and small splint fixation and fumigation with traditional Chinese medicine can significantly improve the hemorheology of the patients and reduce the contents of inflammatory factors in the body, with significant efficacy and no adverse reactions, which is worthy of clinical application.

Background: and Purpose We investigated the causal relationships between the gut microbiota (GM), stroke, and potential metabolite mediators using Mendelian randomization (MR). Methods: We leveraged the summary statistics of GM (n=18,340 in the MiBioGen consortium), blood metabolites (n=115,078 in the UK Biobank), and stroke (cases n=60,176 and controls n=1,310,725 in the Global Biobank Meta-Analysis Initiative) from the largest genome-wide association studies to date. We performed bidirectional MR analyses to explore the causal relationships between the GM and stroke, and two mediation analyses, two-step MR and multivariable MR, to discover potential mediating metabolites. Results: Ten taxa were causally associated with stroke, and stroke led to changes in 27 taxa. In the two-step MR, Bifidobacteriales order, Bifidobacteriaceae family, Desulfovibrio genus, apolipoprotein A1 (ApoA1), phospholipids in high-density lipoprotein (HDL_PL), and the ratio of apolipoprotein B to ApoA1 (ApoB/ApoA1) were causally associated with stroke (all P<0.044). The causal associations between Bifidobacteriales order, Bifidobacteriaceae family and stroke were validated using the weighted median method in an independent cohort. The three GM taxa were all positively associated with ApoA1 and HDL_PL, whereas Desulfovibrio genus was negatively associated with ApoB/ApoA1 (all P<0.010). Additionally, the causal associations between the three GM taxa and ApoA1 remained significant after correcting for the false discovery rate (all q-values <0.027). Multivariable MR showed that the associations between Bifidobacteriales order, Bifidobacteriaceae family and stroke were mediated by ApoA1 and HDL_PL, each accounting for 6.5% (P=0.028) and 4.6% (P=0.033); the association between Desulfovibrio genus and stroke was mediated by ApoA1, HDL_PL, and ApoB/ApoA1, with mediated proportions of 7.6% (P=0.019), 4.2% (P=0.035), and 9.1% (P=0.013), respectively. Conclusion The current MR study provides evidence supporting the causal relationships between several specific GM taxa and stroke and potential mediating metabolites.

OBJECTIVETo review the advances of studies on vitamin D receptor and its role in the pathogenesis of diabetic nephropathy.

DATA SOURCESA comprehensive search of the PubMed literatures without restriction on the publication date was carried out using keywords such as vitamin D receptor and diabetic nephropathy.

STUDY SELECTIONArticles related to vitamin D receptor and diabetic nephropathy were selected and carefully analyzed.

RESULTSThe ligands as well as construction and tissue distribution of vitamin D receptor were summarized. Pathogenesis of diabetic nephropathy was analyzed. The mechanisms underlying the renoprotective role of vitamin D receptor including inhibition of renin-angiotensin system, anti-inflammation, anti-fibrosis and the reduction of proteinuria were reviewed. Mounting evidences from animal and clinical studies have suggested that vitamin D therapy has beneficial effects on the renal systems and the underlying renoprotective mechanisms of the vitamin D receptor-mediated signaling pathways is a hot research topic.

CONCLUSIONOur study suggests that vitamin D receptor has a great potential for preventing the progression of diabetic nephropathy via multiple mechanisms.

Animals ; Diabetic Nephropathies ; metabolism ; Humans ; Proteinuria ; metabolism ; Receptors, Calcitriol ; metabolism ; Renin-Angiotensin System ; physiology

Objective To investigate the clinical effect of acupotomology combined with Bushenzhuanggu prescription in the treatment of residual lumbar pain after percutaneous kyphoplasty(PKP)for osteoporotic vertebral compression fractures.Methods A total of 77 patients with PKP postoperative residual lumbar pain admitted to the Department of Orthopedics,Ningbo Hospital of Traditional Chinese Medicine Affiliated to Zhejiang Chinese Medical University from January 2018 to December 2021 were randomly divided into control group and observation group.Patients in control group were treated with calcitriol soft capsules and calcium carbonate D3 tablets,and patients in observation group were treated with acupotomy combined with Bushenzhuanggu prescription.The pain degree,lumbar function and self-care ability of the two groups were compared.Results After treatment,the pain level,lumbar function,and self-care abilityof the two groups of patients improved compared to before treatment.Compared with control group,the visual analogue score(VAS)score and Oswestry disability index(ODI)index of observation group decreased more significantly,and the difference was statistically significant(P<0.05);The Barthel index of observation group increased more significantly in the early stage(3 days and 1 month after treatment),with a statistically significant difference(P<0.05).However,there was no statistically significant difference in Barthel index between the two groups at 3 months after treatment(P>0.05).Conclusion Acupotomy combined with Bushenzhuanggu prescription has satisfactory clinical efficacy in the treatment of residual lumbar pain after PKP operation of osteoporotic vertebral compression fracture.It can significantly reduce pain,improve lumbar function,shorten the improvement time of patients'self-care ability,improve patients'quality of life,and effectively promote patients'rapid recovery.

OBJECTIVE To explore wheth er there is a relationship between the judgment results of medical damage liability disputes related to off-label drug use and evidence-based evidence. METHODS By searching for medical damage liability disputes related to off-label drug use up to 2021 on pkulaw.cn ，documents were extracted to record objective factors ，subjective factors and judgment results ；whether there was evidence-based evidence was judged according to Off-label Drug Use List and Evidence-based Evaluation Standards for Off-label Drug Use of Guangdong Pharmaceutical Association ；univariate analysis was adopted to test the relationship between the judgment results and evidence-based evidence. RESULTS A total of 57 cases were included. Cases mainly occurred in the eastern China （63.2％）and tertiary hospitals （64.9％），the main appraisal agency was the appraisal center or institute（61.4％），and the most common type of off-label drug use was overdose drug use （45.6％）. Among the judgment results ， 23 cases（40.4％）of off-label drug use had a causal relationship with medical damage ，most of the responsibility of doctors was secondary responsibility （28.1％），and the actual compensation amount of the most cases were less than 100，000 yuan（54.4％）. There were 25 cases（43.9％）with evidence-based evidence. Univariate analysis found that for off-label drug use the claim amount of the case with evidence-based evidence was significantly higher than that of the case without evidence-based evidence （P＝ 0.040），and there was no significant correlation between evidence-based evidence and the actual compensation amount of the case （P＝0.741），causality determination （P＝0.256），liability type （P＝0.598）or appraisal agency （P≥0.260）. CONCLUSIONS There is no significant correlation between the judgment results of medical damage liability disputes related to off-label drug use and evidence-based evidence ，indicating that there may be certain differences between judicial trials and medical science. The off-label drug use should be regulated by establishing a complete off-label drug use management system and standardizing informed consent procedure for off-label drug use. 1610307322@pku.edu.cn