OBJECTIVE:To establish a method for simultaneous determination of andrographlide and dehydroandrographolide in Ganmaoqing tablet. METHODS:HPLC was performed on the column of Thermo BDS C18 with mobile phase of mathanol-water at a flow rate of 1.0 ml/min,detection wavelength was 225 nm for andrographlide and 254 nm for dehydroandrographolide,col-umn temperature was 30 ℃,and injection volume was 10 μl. RESULTS:The linear range was 2.25-45.00 μg/ml for andrographlide (r=0.9998) and 1.00-20.00 μg/ml for dehydroandrographolid(r=0.9990);RSDs of precision,stability and reproducibility tests were no more than 0.8%;average recoveries were 96.96%-99.80%(n=9) and 96.79%-100.65%(n=9). CONCLUSIONS:The method is simple,accurate and reliable,and can be used for the quality control of andrographlide and dehydroandrographolide in Ganmaoqing tablet.

BACKGROUND:Previous research has demonstrated that preoperative hormone therapy relieved clinicaI and pathological stages of prostatic carcinoma patients,reduced positive ratio of incisaI margin:however,it did not increase disease-free surviva J rate.Worth of preoperative hormone therapy for prostatic carcinoma remains unclear.OBJECTIVE:To assess the role of preoperative hormone therapy in localised prostate cancer.METHODS:Articles were searched from PubMed,Embase.Cochrane Library(No.4,2009),Chinese biomedicaf literature database,Chinese Scientific Journals full_text database,and Chinese Journal full-text database(published before October,2009).Randomized controlled trials which were diagnosed as Iocalised prostate cancer using pathology and cytology were included The sex and nationality were not limited.and patients did not have severe heart and lung diseases Prostatic carcinoma which occurred Later or repeatedly was excluded.Otherwise.randomized controlled trails which were coincidence with the inclusion criteria were also included.Review Manager 5.0 published by Cochrane was used for stafistical analysis.Overall survival rate.disease-free survivaI rate,positive surgical margin rate,positive lymph node rate.and seminal vesicle invasion rate were evaluated.RESULTS AND CONCLUSlON:Six randomized controlled trials totaling 1 027 padicipants were included.The result of meta analysis showed that there was no significant difference in overall surviva J rate[RR=0.94,95%C,(0.86,1.02)],disease-free survival rate[RR=1.02,95%CI(0.89,1.17)],positive lymph node rate IRR=0.86,95%CI(0.47,1.57)],and seminal vesicle invasion rate[RR=1.09,95%CI(0.74,1.59)]between single prostatectomy and preoperative hormone therapy plus prostatectomy,while there was significant difference in positive surgical margin rate[RR=0.46,95%CI(0.32,0.66)]This suggested that preoperative hormonal therapy prior to prostatectomy did not improve survival rate,positive lymph nodes rate and seminaI vesicle invasion rate:however.there was a significant reduction in the positive surgicaI margin rate.

To develop simple, rapid, and efficacious diagnostic methods for malaria is one of the remaining key tasks for malaria control. Previously, we have created a phage-displayed antibody library against Plasmodium falciparum. Six clones of antibody with good reactivity to HRP-II in ELISA were isolated from the library after 3 rounds of enrichment. Soluble ScFvs were produced and the characteristics were determined. The results of Western blot showed that they could bind to HRP-II specifically and had a relative molecular mass(Mr) about 31 000. The work provided a solid fund for diagnostic kit development for malaria.

OBJECTIVE: To construct a system of therapeutic effect evaluation for patients with primary liver cancer according to the theory of syndrome differentiation in traditional Chinese medicine (TCM), and to examine its reliability. METHODS: Analytic hierarchy process and 100 mm surveyor's rod method were applied to obtain bottom layer and top level syndromes, which were used to construct the method of therapeutic effect evaluation, and its reliability was verified in clinical practice by comparing with some evaluation criteria in Western medicine, such as cancer severity scale; Karnofsky performance scale; Child-Pugh classification, cancer staging classification, and quality of life scale, etc. RESULTS: A system of therapeutic effect evaluation was constructed, and it could reflect the progress of tumor, changes of hepatic function and constitution. The evaluation scores acquired from the system were highly associated with the quality of life of the patients. CONCLUSION: The system of therapeutic effect evaluation can reflect the severity of disease and the characteristics of TCM treatment.

BACKGROUND: There are many in vitro selection method of sperm, and swim-up and density gradient centrifugation are commonly used. It remains unclear which method minimizes bad stimulation to the sperm and select sperm with high fertilization potential. OBJECTIVE: To evaluate the effectiveness of swim-up and gradient centrifugation preparation techniques on intrauterine insemination (IUI).METHODS: A computer-based online search of Cochrane Library, PubMed, EMBASE databases was performed, and some related journals were manually searched for related articles published between January 1966 and February 2009. The quality of included randomized controlled trials (RCT) and q-randomized trials (Q-RCT) was evaluated and Meta-analysis was conducted by the Cochrane Collaboration's software RevMan5.0. Experts.RESULTS AND CONCLUSION: A total of 6 studies were included, involving 4 RCTs and 2 Q-RCTs. A total of 486 patients (1 099 IUI cycles) were enrolled. The Meta-analysis indicated that there was no difference between swim-up and gradient centrifugation preparation techniques for the IUI in terms of cycle pregnancy rates [OR = 1.11, 95%CI(0.8,1.55)], miscarriage rates [OR = 0.31, 95%CI(0.09,1.04)], sperm count [the weight mean difference (WMD) =-0.89, 95%CI(-14.17,12.38)], sperm motility [WMD = -2.31, 95%CI(-7.27,2.65)]. There is insufficient evidence to confirm which is the best method in the two specific preparation techniques. The quality of study methods should be improved. And more measure parameters should be included when comparing it before or after treatment, such as sperm motility, sperm count, sperm function.

Objective:To analyze clinical data of 3 children with LMX1B-associated disease characterized by asymptomatic glomerular proteinuria, thus improving the recognition of asymptomatic proteinuria with genetic causes. Methods:Three patients with LMX1B-associated disease presented with prominent asymptomatic proteinuria diagnosed by the next-generation sequencing in Department of Pediatrics, Peking University First Hospital from April 2014 to October 2017 were included in this study.Clinical data, including renal and extrarenal manifestations, renal biopsy, and family history, were collected and retrospectively analyzed. Results:All 3 children were girls, the age of onset were 2 years, 1 year, and 4 years, respectively, and the diagnosis age were 11 years, 5 years and 6 years, respectively.All of them had glomerular proteinuria, and nephrotic-level proteinuria occurred in one patient.Microscopic hematuria was found in 2 patients.All of them had normal renal function.Only one patient underwent renal biopsy.Electron microscopy of the first time of biopsy revealed segmental thinning of the glomerular basement membrane.Re-biopsy 4 years later showed irregular thickening of the glomerular basement membrane, moth-eaten appearance and collagen fibrillar material deposition.No abnormalities of nails, limbs and joints were observed by physical examination.Two patients had a family history of renal disease.Conclusions:Genetic factors should be considered in children with obscure onset asymptomatic proteinuria without definite clinical causes.Genetic testing can help diagnose and guide treatment as early as possible.

As one of the key enzymes in cell metabolism, the activity of citrate synthase 3 (CS3) regulates the substance and energy metabolism of organisms. The protein members of CS3 family were identified from the whole genome of apple, and bioinformatics analysis was performed and expression patterns were analyzed to provide a theoretical basis for studying the potential function of CS3 gene in apple. BLASTp was used to identify members of the apple CS3 family based on the GDR database, and the basic information of CS3 protein sequence, subcellular localization, domain composition, phylogenetic relationship and chromosome localization were analyzed by Pfam, SMART, MEGA5.0, clustalx.exe, ExPASy Proteomics Server, MEGAX, SOPMA, MEME, WoLF PSORT and other software. The tissue expression and inducible expression characteristics of 6 CS3 genes in apple were determined by acid content and real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Apple CS3 gene family contains 6 members, and these CS3 proteins contain 473-608 amino acid residues, with isoelectric point distribution between 7.21 and 8.82. Subcellular localization results showed that CS3 protein was located in mitochondria and chloroplasts, respectively. Phylogenetic analysis divided them into 3 categories, and the number of genes in each subfamily was 2. Chromosome localization analysis showed that CS3 gene was distributed on different chromosomes of apple. The secondary structure of protein is mainly α-helix, followed by random curling, and the proportion of β-angle is the smallest. The 6 members were all expressed in different apple tissues. The overall expression trend from high to low was the highest relative expression content of MdCS3.4, followed by MdCS3.6, and the relative expression level of other members was in the order of MdCS3.3 > MdCS3.2 > MdCS3.1 > MdCS3.5. qRT-PCR results showed that MdCS3.1 and MdCS3.3 genes had the highest relative expression in the pulp of 'Chengji No. 1' with low acid content, and MdCS3.2 and MdCS3.3 genes in the pulp of 'Asda' with higher acid content had the highest relative expression. Therefore, in this study, the relative expression of CS3 gene in apple cultivars with different acid content in different apple varieties was detected, and its role in apple fruit acid synthesis was analyzed. The experimental results showed that the relative expression of CS3 gene in different apple varieties was different, which provided a reference for the subsequent study of the quality formation mechanism of apple.
Citric Acid ; Malus/genetics* ; Citrate (si)-Synthase ; Phylogeny ; Citrates

Pyruvate dehydrogenase E1 component subunit beta-1 (PDHB-1) is a gene encoding the β-subunit of pyruvate dehydrogenase complex, which plays an important role in fruit acid accumulation. The aim of this study was to investigate the evolution characteristics of apple PDHB-1 family and its expression in apples with different acid contents. Bioinformatics analysis was performed using databases including NCBI, Pfam and software including ClustalX, MEGA, and TBtools. By combining titratable acid content determination and quantitative real-time PCR (qRT-PCR), the expression of this family genes in the peel and pulp of apple 'Asda' and 'Chengji No.1' with different acid content were obtained, respectively. The family members were mainly located in chloroplast, cytoplasm and mitochondria. α-helix and random coil were the main factors for the formation of secondary structure in this family. Tissue-specific expression profiles showed that the expression of most members were higher in fruit than in other tissues. qRT-PCR results showed that the expression profile of most members was consistent with the profile of titratable acid contents. In the peel, the expression levels of 14 members in 'Asda' apples with high acid content were significantly higher than that in 'Chengji No.1' apples with low acid content, where the expression difference of MdPDHB1-15 was the most significant. In the pulp, the expression levels of 17 members in 'Asda' apples were significantly higher than that in 'Chengji No.1' apples, where MdPDHB1-01 was the most highly expressed. It was predicted that PDHB-1 gene family in apple plays an important role in the regulation of fruit acidity.
Malus/metabolism* ; Fruit/genetics* ; Protein Structure, Secondary

ObjectiveTo investigate the late detection of HIV/AIDS cases in the elderly in Jilin Province and analyze its influencing factors， to provide theoretical basis for improving their life quality. MethodsThe first CD4 values of HIV/AIDS patients aged 50 years and above living in Jilin Province were used to estimate late detection， and the influencing factors of late detection in elderly cases were analyzed. ResultsThe average CD4 cell count of newly reported HIV/AIDS cases aged 50 and above in Jilin Province from 1996 to 2021 was （230.55±191.97）， the low value group accounted for the largest proportion （50.8%）， and the late detection rate was 59.3% （1397/2325）. The late detection cases were mainly from sexual transmission （46.8% for same-sex and 48.2% for heterosexual contact）. From the perspective of sample sources， most of the late detection patients were diagnosed while testing for other illnesses， followed by testing and consulting. In terms of contact history， the late detection of cases of men who have sex with men was higher. The binary logistic regression analysis showed that gender， marriage， sample source and report year were the factors affecting the late detection of AIDS. The late detection rate of males was higher， and cases of married couples were more likely to be late detection. With the increase of report year， the late detection rate decreased， and testing and counseling could effectively reduce the late detection rate of AIDS. ConclusionThe CD4 cell count in the first detection of HIV/AIDS in the elderly in Jilin Province is low， and the late detection rate of male cases is high. In recent years， the expansion of voluntary counseling and testing in Jilin Province has effectively reduced the late detection rate of HIV/AIDS. At the same time， sex education should be strengthened for the elderly， healthy marital relationships should be advocated and more attention should be paid to the mental health of the elderly.

Objective : To explore the type and control measure of black dot⁃like contaminants in cell cultures. Methods: The growth state of bacteria was investigated under an inverted microscope ; Their morphological characteristics were analyzed by Gram and auramine O staining as well as electron microscopy; 16S rDNA gene sequencing was used to analyze bacterial species ; Drug sensitivity test was used to screen antibiotics against the bacteria;Cryopreserved SH⁃SY5Y cells were resuscitated by cell culture supernatant of RAW264 cells. Results: Inverted microscopic real⁃time observations showed that black dot⁃like substances had two growth states : static and moving.They were negative for Gram staining while positive for auramine O staining. Electron microscopy revealed that they were short rod⁃shaped bacteria with a polar flagellum during moving phase. 16S rDNA gene sequencing showed that these bacteria were phenylobacterium zucineum HLK1. Ceftriaxone , carboxycillin and imipenem were screened by drug sensitivity test to have inhibitory effects on the bacteria , but cell culture experiments showed that they could not remove the bacteria from SH⁃SY5Y cells. Contaminated cells could not be cryopreserved for a long time , but resuscitation with RAW264. 7 cell culture supernatant significantly improved the survival rate of cells. Conclusion The black dot⁃like contaminants in cell cultures are a special type of oligotrophic bacterium with strong viability that can invade the cells and cannot be cleared with antibiotic treatment. RAW264. 7 cell culture supernatant seems contain some substances against bacteria , and resuscitating frozen cells with RAW264. 7 cell culture supernatant may significantly improve the survival rate of cells.