Objective:To establish a determination method of Bergenin content in Dicha Kechuan oral liquid by HPLC.Methods:The separation was performed on SHIMADZU VP-ODS C18 column(150mm?4.6mm,5?m),the mobile phase was carbinol-water(20:80) with a flow rate of 1.0ml/min.The column temperature was room temperature and the detection wavelength was 275nm.Results:The linear range of Bergenin was 0.2575-5.150?g,the regression equation of working curve was Y=812702.2X-80626.3,r=0.9998,the average recovery was 101.4% and RSD 1.67%(n=5).Conclusion:This method is simple,reproducible,and can be used for quality control of Dicha Kechuan Oral Liquid and the semifinished product.

Objective: To construct a liver targeting gene transfer vector using hepatitis B virus envelope particles. Methods: Hepatitis B viruses were obtained from the supernatant of HepG 2.2.15 cells by a PEG8000 system and were inactivated by ?-propiolactone to prepare hepatitis B virus envelope. The hepatitis B virus envelope was used to pack 5.3 kb pIRES_2-EGFP to assess their packing ability. Subsequently, the products were studied with ELISA, PCR, SDS-PAGE, and electron microscopy. Finally, the product was used to transfect HepG2 cells and the green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer.Results: The acquired hepatitis B virus envelope retained the surface protein HBsAg+pre S_1+pre S_2, but with no virus DNA. The prepared envelope had high packing ability for GFP and the packed GFP had a high transfection rate in HepG2 cell. Conclusion: Hepatitis B virus envelope has been successfully obtained from the supernatant of HepG 2.2.15 cells with a PEG8000 system and ?-propiolactone.

[Objective] To learn about the genetic diversity,we studied the five X-chromosomal STR (X-STR) loci in Guangdong Han Nationality Groups.[Methods] The five Loci (DXS6803,DXS981,DXS6809,DXS6789,and DXS7132) were amplified in a pentaplex PCR reaction.PCR products were analyzed using capillary electrophoresis and ABI prism 3100 Genetic Analyzer,with GeneMapper ID 3.1 Analysis Software.[Results] A total of 363 individuals (181 unrelated male and 182 unrelated female) from Guangdong Han population were tested,54 alleles were observed for these loci.Polymorphism information content is 0.6935 ～ 0.8177.Power of discrimination in females was 0.8976 ～ 0.9562.Mean exclusion chance for X-STR in standard trios with daughters was 0.7805 ～ 0.8467.[Conclusion] The five loci in the multiplex system provide high polymorphism information for forensic identification and paternity testing,particularly for difficult paternity deficiency cases.

Objective: To observe the promoting blood circulation by removing blood stasis of Lubai Capsule(LBC)(Rhizoma Phragmitis, Radix Paeoniae Alba, Radix Saposhnikoviae, Flos Schizonepetae, etc.). Methods: Acute blood stasis rat models were established with swimming in iced water and sc adrenalin in order to observe the effect of LBC on blood rheology. Mesenteric microcirculatory disturbance rat models were also established with adrenalin in order to observe the effect of LBC. Clotting time was measured in vitro with prothrombin time(PT) and kaolin partial thromboplastin time(KPTT) kit in order to observe its effects. Results: LBC could decrease the whole blood and plasma viscosity, fibrinogen, erythrocyte sedimentation and aggregation ratio of blood platelets of rats, ease the sticky condition of blood stasis rat models and prevent from forming thrombus. It could also inhibit the constraction and slowing of blood flow of thin artery, the reducing of open capillaries and change of fluid condition caused by adrenalin and improve these phenomena. PT and KPTT could be increased obviously. Conclusion: LBC can significantly promote blood circulation by removing blood stasis, because of improving blood rheology and mesenteric microcirculatory disturbance and inhibit endogenous and exogenous coagulation system.

Objective To design and construct a new non-fusion soluble expression vector pTIG-mSUMO(small ubiq-uitin-related modifier) using the widely used solubility promoting protein SUMO and based on the translational coupling phenomenon in order to enable the non-fusion soluble expression of the broad-spectrum antiviral protein RA in Escherichia coli by pTIG-mSUMO.Methods The smt3 gene coding for SUMO protein was cloned from yeast genome DNA by PCR. After directed-site silent mutation to eliminate the EcoRⅠsite, the mutant mSUMO was inserted into pET-22b to obtain the translational coupling expression vector pTIG-mSUMO.The RA was subject to PCR amplification and cloned into the pTIG-mSUMO to obtain the expression plasmid pTIG-mSUMO/RA which was supposed to direct the soluble expression of RA by the translational coupling with mSUMO.Results A translational coupling expression vector pTIG-mSUMO which could di-rect/drive the SUMO and heterogonous protein non-fusion expression simultaneously was designed and constructed.The Western blotting result indicated that pTIG-mSUMO could direct the high-level expression of RA, around 40%of which was soluble.Conclusion A translational coupling expression vector pTIG-mSUMO is obtained.After coupling with SUMO, RA is highly expressed in E.coli and both the expression level and solubility are greatly improved.pTIG-mSUMO might contrib-ute to soluble expression of other proteins.

OBJECTIVETo establish a nine X-chromosome short tandem repeats (X-STR) loci multiplex PCR method and study the polymorphism of the 9 X-STR loci,and to determine its application in kinship tests for forensic medicine.

METHODSA fluorescent multiplex PCR that simultaneously amplifies 9 X-STR loci, i.e. DXS7133, DXS981, DXS7424, DXS6789, DXS7132, GATA165B12, DXS101, GATA31E08 and DXS10011, were set up. PCR products were analyzed using capillary electrophoresis and ABI prism 3100 Genetic Analyzer with GeneMapper ID 3.1 analysis software.

RESULTSWhen 251 unrelated male and 112 unrelated female individuals from southern China were tested, 111 alleles were detected. The power of discrimination in females was 0.5837-0.9959. Mean exclusion chance for X-STR in standard trios with daughters was 0.4072-0.9511. Polymorphism information content was 0.4481-0.9531.

CONCLUSIONThe results demonstrate that the 9 loci in the multiplex system provide high polymorphism information, and the multiplex system provides a fast technology for forensic identification and paternity testing. The X-STR multiplex system can complement the analysis of AS-STR and Y-STR efficiently.

Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Chromosomes, Human, X ; genetics ; Female ; Humans ; Male ; Microsatellite Repeats ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic

OBJECTIVETo investigate the association between rs185983011 single-nucleotide polymorphisms (SNP) of apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3G (APOBEC3G) and the susceptibility to chronic hepatitis B.

METHODSThe blood samples were collected from 186 healthy subjects and 159 patients with chronic hepatitis B. The rs185983011 SNP was detected and genotyped by sequencing with Sanger's method to analyze the relationship between rs185983011 SNP and chronic hepatitis B.

RESULTSOnly C/C and C/T genotypes of the alleles of rs185983011 SNP were found in the tested subjects, and the C/C genotype was predominant (97.7%). The distribution frequencies of rs185983011 SNP genotypes and alleles showed no significant difference between healthy subjects and patients with chronic hepatitis B (P>0.05).

CONCLUSIONThe predominant genotype of rs185983011 SNP of APOBEC3G is C/C in the tested subjects, and rs185983011 SNP does not appear to associate with the susceptibility to chronic hepatitis B.

APOBEC-3G Deaminase ; Adult ; Alleles ; Case-Control Studies ; Cytidine Deaminase ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Hepatitis B, Chronic ; genetics ; Humans ; Male ; Polymorphism, Single Nucleotide ; Young Adult

Purpose/Significance The paper systematically reviews the application and research progress of large language models(LLMs)in the medical field,analyzes the key challenges and opportunities,and provides references for related research.Method/Process The systematic literature review method is adopted to comprehensively review the relevant literature published in recent years,fo-cusing on the latest progress of medical LLMs.The application results,research status and challenges of LLMs in medical natural lan-guage processing(NLP)tasks are analyzed.Result/Conclusion The application of LLMs in the medical field has a broad prospect,and the future research should focus on the technical progress and the improvement of ethical norms.On the one hand,the pace of technologi-cal innovation should be accelerated,and on the other hand,strict compliance with ethical standards should be ensured to jointly promote the sustainable development of LLMs technology in the medical field.

OBJECTIVETo establish a one-step four multiplex reverse transcription polymerase chain reaction (RT-PCR) method based on Homo-Tag Assisted Non-Dimer System (HAND) system for simultaneous detection of 4 diarrhea viruses of rotavirus, astrovirus, norovirus and sapovirus.

METHODSPrimers were designed according to the conserved genome sequence of the 4 viruses and the homologous tail sequences were added to the 5' end. The multiplex RT-PCR system was constructed by optimizing the PCR parameters such as the concentration of universal tag primer and genome-specific Homo-Tailed primers. The specificity, stability and sensitivity of the method were evaluated systematically.

RESULTSThe 4 multiplex RT-PCR methods based on HAND system was established successfully. Specificity analysis showed no cross reaction between the 4 diarrhea viruses. The sensitivity analysis showed detection limits for rotavirus, astrovirus, norovirus and sapovirus of 48, 1.92, 9.6 and 48 pg per reaction, respectively.

CONCLUSIONThe established HAND system-based multiplex RT-PCR assay allows simple, rapid, specific, sensitive, and stable for detection of the 4 common diarrhea viruses at low costs and is suitable for application in general medical laboratories.

Astroviridae ; genetics ; isolation & purification ; Diarrhea ; virology ; Feces ; virology ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Norovirus ; genetics ; isolation & purification ; RNA, Viral ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Rotavirus ; genetics ; isolation & purification ; Sapovirus ; genetics ; isolation & purification

Necrosis is a form of cell death, which is related to various serious diseases such as cardiovascular disease, cancer, and neurodegeneration. Necrosis-avid agents (NAAs) selectively accumulated in the necrotic tissues can be used for imaging and/or therapy of related diseases. The aim of this study was to preliminarily investigate necrosis avidity of I-evans blue (I-EB) and its mechanism. The biodistribution of I-EB at 24 h after intravenous administration showed that the radioactivity ratio of necrotic to viable tissue was 3.41 in the liver and 11.82 in the muscle as determined by counting in model rats. Autoradiography and histological staining displayed preferential uptake of I-EB in necrotic tissues. nuclear extracts from necrotic cells exhibited 82.3% of the uptake in nuclei at 15 min, as well as 79.2% of the uptake at 2 h after I-EB incubation. The DNA binding study demonstrated that evans blue (EB) has strong binding affinity with calf-thymus DNA (CT-DNA) (=5.08×10 L/(mol/L)). Furthermore, the accumulation of I-EB in necrotic muscle was efficiently blocked by an excess amount of unlabeled EB. In conclusion, I-EB can not only detect necrosis by binding the DNA released from necrotic cells, but also image necrotic tissues generated from the disease clinically.