Objective To study the Application of X-linked differentially methylated polymorphism site in forensic medicine.Methods X-STR HUMARA was chosen as a model locus.PCR procedures were performed after digestion using methylation-sensitive restriction endonucleases.STR polymorphism of HUMARA was analyzed and compared in samples collected from male and female individuals.Result After digestion with methylation-sensitive restriction enzyme HpaⅡ,no PCR products were obtained from male samples,whereas PCR products from the female samples were normally typed.In monoclonal tumor cell samples from females,only one allele was detected.Conclusion The differentially methylated X-STR HUMARA locus is a novel marker for mixture analysis of mixed stains,sex determination and discrimination of tumor tissues.

Drowning represents one of the leading causes of unnatural death. The plankton test is still considered as one of the useful methods for medico-legal investigation of death by drowning although controversies concerning the reliability of the test exist. The methods commonly used for detection of planktons generally include microscopic examination after tissue samples are destructed with or without strong acid, and detection of the planktons by molecular biological techniques. The evaluation of the positive results by the former methods are limited by the physical-chemical characters of the planktons. Detection of DNA genetic markers by molecular biological techniques, which can provide abundant information and is applicable to studying various planktons, is a novel method helpful for investigation of drowning. In the present paper, several techniques for detection of planktons were reviewed as references for medico-legal practitioners in investigation of suspected cases of drowning.

HIV-1 integrase (IN) is an essential enzyme for retroviral replication. There is no analogue for this enzyme in human cells so that inhibition of IN will not bring strong effect on human body. Thus, HIV-1 IN has become a rational target for therapy of AIDS. This review provides a comprehensive report of alpha, gamma-diketo IN inhibitors discovered in recent years. Compilation of such data will prove to be beneficial in developing QSAR, pharmacophore hypothesis generation and validation, virtual screening and synthesis of compounds with higher activity.

Objective To explore the feasibility of hemochrome identification by micro spectrophotometer. Methods We use dip lotion by Centrifugation, reaction with modiifed takayama reagent,measured absorption spectra with a micro spectrophotometer to determine whether samples contain blood. On the basis of optimized parameters, we use diluted human hemoglobin to measure sensitivity of the method; using suspected blood / blood spots, and different storage time and matrix of blood spots to measure speciifcity and samples of adaptability. Result With micro spectroscopic method of hemochromogen,only blood (spot) has speciifc spectral consisting of three absorption peaks at 415,525 and 555nm and no cross suspected common blood / spot.Reaction 2 min, sample volume2.5ul, 1000-fold diluted human blood stably obtains primordial spectrum. By prolonging the immersion time, set the vehicle control, 10 years old blood gauze. Blood stains in colored cloth can be effective detective.. The height of absorption peaks and blood content were significantly corelated, and the change in absorbance at 525 and 555nm consistent trend (y=0.5232x+0.0274, R2=0.9971). Conclusion Micro spectroscopic method of hemochromogen has high sensitivity and speciifcity, quick and easy, can be incorporated into the DNA testing process. There is a good prospect in the actual seizure case. Instead of the traditional crystallization method for teaching, training requirements more in line with the skills and awareness for current students.

Objective To explore the feasibility of "coffee ring" effect in DNA high-throughput evaluation. Methods we mixed uniformly DNA solution and EB solution, then dripped different size of droplets on 5 kinds of substrates. After evaporation, the coffee ring was observed and photographed with LG2020 gel imaging analysis system. Analyzed the correlation among ring formation conditions,ring fluorescence intensity and DNA concentration, and looked for the sensitivity and applicable range of the method. Result DNA - EB solution can form fluorescent ring after surface evaporation on the slide, transparent polystyrene plate carrier and so on. The DNA detection limit can be up to 2ng/μl. DNA concentration and loop integral optical density fitting Logistic equation(R2=0,999) within 2ng/μl~1000ng/μl. Conclusion The coffee ring method is simple, fast and low equipment requirement,which can be used for high-throughput evaluation of DNA solution.

Objective To develop a simplified bisulfite genomie sequencing(BGS)method for DNA methylation marker scanning.Methods According to modified BGS protocol,the desalt DNA treated with bisulfite were directly used for bisulfite-PCR(BSP)without alkali treatmenL Complement of the bisulfite modification Wag accomplished by a prolonged pre-denaturation stage.After BSP,a second round PCR was performed with a pair of GC tagged primers to adjust the GC content of the amplieon for direct sequencing.To assess this improved protocol,promotor methylation of TNF-α gene in 3T3-L1 cell and androgen receptor(AR)gene in Hela cell was investigated.The real time BSP for Alu was also used to compare the sensitivity of the modified assay with traditional assay.Results Both the hypermethylated TNF-α promotor and hypomethylated AR promotor were successfully sequenced by improved BGS method,and the results were consistent with that of the traditional assay.The conversion rate reached 100%,while the conversion specificity was higher than 93.75%.The sensitivity of improved BGS method inereaged significantly(t=2.978 2,P＜0.05)and showed good reproducibility.Condusion The improved BGS method is simple and sensitive,facilitating more ambitious genomic methyhtion mapping studies.

Objective To investigate the influences of drop components on the"coffee ring" test of DNA. Methods The DNA-EB drops containing SDS, NaCl, etc, were evaporated on glass slides. After evaporation, the fluorescent Deposition Pattern (DP) and Integrated Optical Density (IOD) was acquired with a gel imaging system. The dose-effect relationship was analyzed. Result When the non-DNA components concentration is low, the DP was still characteristiced by peripheral rings, subtle component-specific differences, such as tree-ring like structure and radial crack, existed. At high concentrations, DP was various, which may be ring + scattered dots (NaCl), central spot + small weak ring (SDS), concentric/tree-ring (TritonX-100) or ring + spot (H+), et al. Non-DNA components had little effect on IOD(0.5~2 times). Conclusion Non-DNA components in DNA-EB drops influences both the DP and IOD, but rough estimation of DNA concentrations is still effective. Moreover, analyzing the DP can provide more informations about sample purity and residue.

Objective:To evaluate the effectiveness and safety of coronavirus disease 2019 (COVID-19) vaccines for dialysis patients with chronic kidney disease.Methods:PubMed, Medline, Embase databases and CNKI, VIP, Wanfang databases were searched systematically. The deadline was April 25, 2022. The search terms included haemodialysis, peritoneal dialysis, vaccine, seroresponse, COVID-19, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The main outcome included the positive rate after vaccination, antibody titer, antibody changes during follow-up, infection rate of SARS-CoV-2, hospitalization rate and mortality.Results:A total of 154 195 patients were analyzed in 26 studies. The results of meta-analysis showed that the positive rate of serum IgG antibody in patients with chronic kidney disease was 48% after the first dose of vaccine, 89% and 96% after the second dose and third dose, respectively. After vaccination with COVID-19 vaccines, there was no significant difference in serum antibody and titer between hemodialysis patients and peritoneal dialysis patients. However, compared with the healthy control group, the antibody positive rate and antibody titer of dialysis patients after vaccination were lower (both P<0.05). In the follow-up, the antibody positive rate at the third month decreased by 12% compared with at the first month, at the sixth month decreased by 15% compared with at the third month, and at the sixth month decreased by 20% compared with at the first month. The serum antibody positive rate after the third dose of vaccine increased by 38% ( RR=1.38, 95% CI 1.12-1.70, P<0.001), and the antibody titer increased significantly ( SMD=1.46, 95% CI 0.31-2.61, P<0.001). Although the vaccines could not reduce the infection rate of SARS-CoV-2 in dialysis patients, it could significantly reduce the hospitalization rate and mortality after infection. Conclusions:After vaccination with COVID-19 vaccines, dialysis patients can produce strong serum antibodies, which can reduce the hospitalization rate and mortality after SARS-CoV-2 infection. However, the duration of antibody is short and the titer level is low, so it is necessary to timely vaccinate booster vaccine dose to obtain stronger immunogenicity.

ObjectiveTo explore the therapeutic effect and mechanism of Yiqi Huoxue Tongbian prescription on slow transit constipation （STC） in rats. MethodThe rat model of STC was established by gavage of loperamide hydrochloride. Rats were assigned into control， model， mosapride， low-， medium-， and high-dose （3.51， 7.02， and 14.04 g·kg^-1， respectively） Yiqi Huoxue Tongbian prescription groups. The changes of general signs， fecal moisture content， and intestinal propulsion rate were measured after model establishment and drug administration. The colonic mucosal changes were observed by hematoxylin eosin staining. Enzyme-linked immunosorbent assay was employed to determine the content of substance P （SP） and vasoactive intestinal peptide （VIP） in the colon of rats in each group. The gray values of aquaporin （AQP） 3， AQP4， AQP8， and c-Kit in rat colon tissue were measured by immunohistochemistry and Western blot， and the changes of intestinal flora were detected by 16S rRNA high-throughput sequencing. ResultCompared with the model group， 10 days of treatment with Yiqi Huoxue Tongbian prescription increased the fecal moisture content and intestinal propulsion rate （P<0.01）. The medium- and high-dose Yiqi Huoxue Tongbian prescription groups and the mosapride group showed no obvious mucosal inflammation and neat arrangement of goblet cells with a large number in the colon tissue. Moreover， the three groups showed increased SP content （P<0.01） and decreased VIP content （P<0.01） in the serum. The medium- and high-dose Yiqi Huoxue Tongbian prescription groups showed down-regulated protein levels of AQP3， AQP4， and AQP8 （P<0.01） and up-regulated protein level of c-Kit （P<0.01）. The drug administration groups presented slightly increased observed species， Chao1， ACE， and Shannon， Simpson， and PD whole tree. The principal component analysis showed that the control group had a short distance with the high- and medium-dose Yiqi Huoxue Tongbian prescription groups， indicating that high- and medium-dose Yiqi Huoxue Tongbian prescription can recover the intestinal flora to that in the control group. ConclusionYiqi Huoxue Tongbian prescription can alleviate the defecation status of rats with slow transit constipation by down-regulating the expression of AQP3， AQP4， and AQP8 to reduce the absorption of water in the intestine， up-regulating the expression of c-Kit to increase the number and distribution of Cajal interstitial cells， and regulating the balance of flora in the colon tissue.

Anoctamin 1 (ANO1) is a kind of calcium-activated chloride channel involved in nerve depolarization. ANO1 inhibitors display significant analgesic activity by the local peripheral and intrathecal administration. In this study, several thiophenecarboxylic acid and benzoic acid derivatives were identified as novel ANO1 inhibitors through the shape-based virtual screening, among which the 4-arylthiophene-3-carboxylic acid analogues with the best ANO1 inhibitory activity were designed, synthesized and compound