An experimental study was carried out on the isolated bone cells, from the embryonic calvarium of Sprague-Dawley rats which were taken out in vitro culture in the later period of gestation. Observation of the early stage of culture was carried out with phase contrast microscope, H. E. staining and AcP and AlP technique. It was found that both the osteogenic cells of the cambium layer of the periosteum and the undifferentiated cells from the bone marrow could develop into large squamous-shaped cells. These cells possessed many processes connected with the adjacent cells and stained light red color with Sirius Red but revealed no birefringency under polarized microscope. This indicates that these cells are precursors of collagen formation. The negative reaction with the alkaline phosphatase staining pointed out that these cells still belong to the osteogenic cells.

Objective To observe the therapeutic effect of trimetazidine on patients with ischemic cardiomyopathy heart failure and the changes of plasma level of N-terminal pro-B type natriuretic peptide(NT-proBNP).Methods 62 patients with ischemic cardiomyopathy heart failure were randomly assigned to regular treatment group(3 1)and trimetazidine treatment group(31).All of them were given either regular medicine treatment or trimetazidine for 4 weeks.Concentration of NT-proBNP and echocardiography were detected before and after treatment and the correlation between NT-pmBNP and ultra-sound values was studied.Results There was no statistic difference in the NT-proBNP level between two groups before treatment,but after 4-week treatment the NT-proBNP level in both groups were deCreased(P＜0.05),the decline in the trimetazidine group was more significant.The NT-proBNP level was significantly correlated to LVEF and LVEDD(γ=-0.472、0.45 1;P＜0.05).Conclusions Trimetazidine has a positive effect on patients with ischemic cardiomyopathy heart failure,and decreases the plasma NT-proBNP level.NT-proBNP Can be used as a follow-up index.

One strain of Hantavirus(HV) was isolated from patients with hemorrhagic fever with renal syndrome (HFRS) in Zhejiang province and its S-segment was cloned and submitted to nucleotide sequence analysis in order to determine the type of strain and extent of genetic variation for further study on its evolution and mutation .The HV antigen was detected from mouse lungs in endemic area by direct immunofluorescene test and the HV-positive sample of mouse lung was inoculated to Vero-E6 cells to isolate virus. The total cellular RNA was extracted from infected cell culture and the S segment gene was amplicated by RT-PCR. Then, the purified PCR product was cloned into T vector for sequencing. The result showed that the full-length sequence of the S segment was 1 700 bp with one open reading frame (ORF) encoding 429 amino acids. Comparison with Hantavirus HTN type showed 85.7%-91.9% homology at the nucleotide level. In comparison with the SEO type of viruses, the homology at nucleotide level was shown to be 71.2%-75%. This new HV strain may be typed as to HTN virus and may be a new subtype of this virus.

Objective To study the complete genome sequence of Hantavirus ZT71 strain gene isolated in Zhejiang province and explore its evolution. nethods The total RNA was prepared from ZT71 virus infected cells and the RT-PCR products were cloned into T vector, sequenced and analyzed. Results The L, M and S segments of the strain ZT71 genome were 6530,3651 and 1753 nucleotides in length with a single open reading frame individually encoding 2151,1133 and 429 amino acids. The sequence analysis of nucleotides showed that the homology of L, M and S segments of strain ZT71 between those of other strains of Seoul virus could reach 95.5%-99.7%, 84.1%-99.6% and 88.7%-99.5%, respectively. The analysis of the deduced amino acids showed the similar result. The source of strain ZT71 could be traced from the analysis of the phylogentic trees of nucleotides and amino acids, and it should belong to Seoul type of Hantavirus which was also verified serologically. Conclusion The nucleotide sequences and deduced amino acid sequences of L, M and S segments of strain ZT71 are similar to that of those of Seoul type of Hantavirus. And Hantaan type virus used to be prevalent primarily in Zhejiang province,and it would be an endemic area of mixed type of Hantavirus since the discoveries of the viruses of Soeul type in recent years.

The M and S segment cDNAs of hantavirus Z5 strain was amplified by RT-PCR,and the purified PCR products were cloned into vector pGEM-T and then sequenced.It was demonstrated that the M genome segment of Z5 was found to be 3 616 nucleotides in length with a single open reading frame encoding 1 135 amino acids.And the S genome segment was 1700 nucleotides in length with a single open reading frame encoding 429 amino acids.As demonstrated by the homologous analysis of nucleotides and amino acids,it was showed that the Z5 strain belonged to hantaan viruses HTN type and was the same subtype of the Z10 strain.It is conclouded that difference in nucleotide sequence exists between Z5 strain with other Hantavirus strains but high level of homology in amino acid sequences is still present.

Objective To investigate the influence of metformin on metabolic syndrome in patients with coronary artery disease.Methods 52 patients with coronary artery disease and metabolic syndrome were randomly assigned into 2 groups.the control group with 20 cases and the study group with 32 cases.All the patients were treated with standard coronary artery disease therapy and meanwhile the study group was administered with metformin in addition to the basic therapy above.The follow-up time is one year.FPG,2h postprandial glucose,FINS,TG,TC,HDL-C,insulin resistance index and other standards were compared before and after the therapy,and between the 2 groups when the observation was finished.The rate of cardiovascular events was also compared between the 2 groups.Results Compared with the control group,the study group had lower levels of FPG,2h postprandial glucose,FINS,TG and insulin resistance index when the therapy was completed(P<0.05),and the rate of cardiovascular events was also lower(P<0.05).Conclusion Metformin could lower the levels of glucose,FINS,BMI,TG and insulin resistance index in patients with coronary artery disease and metabolic syndrome.Thus,it could reduce and undermine cardlovascular risk factors,protect the cardiovascular system,reduce the rate of cardiovascular events,which fully benefits the patients.

Single-cell RNA sequencing is a method for transcriptome profiling at the single-cell level,and is a hotspot technology in the field of biological research currently.Compared with the common high-throughput RNA sequencing,single-cell RNA sequencing can distinguish the biological differences between single cells,and identify rare cell populations and subpopulations.This review mainly describes the single-cell RNA sequencing method,summarizes current application of this technique in the field of dermatology,and discusses limitations and prospects of this technique.

Objective:To study the influence of time-dependent acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score on 14-day death risk in patients with severe stroke, and to provide reference for clinical diagnosis and treatment.Methods:Data of 3 229 patients with severe stroke were enrolled from Medical Information Mart for Intensive Care-Ⅲ (MIMIC-Ⅲ). According to the main types of stroke, the patients were divided into subarachnoid hemorrhage (SAH), intracerebral hemorrhage (ICH), ischemic stroke (IS) and other groups. According to age, patients were divided into > 60 years old and ≤ 60 years old subgroups. According to the baseline of sequential organ failure assessment (SOFA) score, they were divided into subgroups of > 3 and ≤ 3. The daily measured values of APACHE Ⅱ scores in each patient were recorded. And all-cause death within 14 days after admission to intensive care unit (ICU) was used as the outcome index to obtain the survival status and survival time of patients. Joint models for longitudinal and time-to-event data were established to evaluate the effect of APACHE Ⅱ score measured at multiple time points on the death risk of patients, and a subgroup analysis was performed.Results:Among the joint models, the one which include APACHE Ⅱ score, and the interaction items between APACHE Ⅱ and age showed the better fitting. Further analysis showed that APACHE Ⅱ score was affected by age, gender, hospital admission, baseline SOFA score and smoking history. After controlling for these confounding factors, APACHE Ⅱ score was significantly associated with 14-day all-cause death in patients with severe stroke [hazard ratio ( HR) = 1.48, 95% confidence interval (95% CI) was 1.31-1.66, P < 0.001], which indicated that the risk of death increased by 48% (95% CI was 31%-66%) for each 1-point increase in APACHE Ⅱ score. Subgroup analysis showed that for different types of severe stroke patients, APACHE Ⅱ score had a greater impact on the risk of 14-day death in SAH patients ( HR = 1.43, 95% CI was 1.10-1.85), but had a smaller impact on ICH and IS groups [HR (95% CI) was 1.37 (1.15-1.64) and 1.35 (1.06-1.71), respectively]. There was no significant difference in APACHE Ⅱ score on the risk of 14-day death between the patients aged > 60 years old and those aged ≤ 60 years old [ HR (95% CI): 1.37 (1.08-1.72) vs. 1.35 (1.07-1.70), respectively]. Compared with patients with SOFA score > 3, APACHE Ⅱ score had a greater impact on the risk of 14-day death in patients with SOFA score ≤ 3 [ HR (95% CI): 1.40 (1.16-1.70) vs. 1.34 (1.16-1.55)]. Conclusion:Time-dependent APACHE Ⅱ score is an important indicator to evaluate the risk of death in patients with severe stroke.