0.05) and the mean operative bleeding (135.8?62.9)ml and (245.1?168.9)ml (P

Objective Dexmedetomidine is known to have a neuroprotective effect.The aim of this study was to investigate the effects of dexmedetomidine on ketamine-induced apoptosis of primarily cultured cortical neurons and its action mechanisms. Methods Rat cortical neurons were primarily cultured for 7 days and treated with ketamine (100μmol/L) and different concentrations of dexmedetomi-dine (0.001, 0.01, 0.1, and 1 μmol/L) for 24 hours, followed by measurement of the viability of the neurons by MTT assay.The neurons were divided into four groups:vehicle control, ketamine ( trea-ted with 100 μmol/L ketamine), dexmedetomidine+ketamine (DD+K, treated with 0.1 μmol/L DD and 100 μmol/L ketamine), and LY294002 ( treated with 0.1 μmol/L DD, 100 μmol/L ketamine, and 10 μmol/L LY294002) .After 24 hours of treatment, the apoptosis rate of the neurons was determined by Hoechst33258 staining, and the expressions of pAkt and cleaved-caspase-3 in the neu-rons detected by Western blot. Results The apoptosis rate of neurons was dramatically increased in the LY294002 and ketamine groups in comparison with the vehicle control and DD+K groups ([36.8 ±4.4] and [43.4 ±4.5]%vs [7.5 ±1.1] and [16.4 ± 3.6]%, P<0.01), the pAkt level remarkably decreased (0.26 ±0.02 and 0.15 ±0.01 vs 0.61 ±0.05 and 0.50 ±0.04, P<0.01), and the expression of cleaved caspase-3 significantly upregulated in the former two as compared with the latter two groups (0.40 ±0.02 and 0.65 ±0.03 vs 0.10 ±0.02 and 0.12 ±0.01, P<0.01). Conclusion Dexmedetomidine exerts a neuroprotec-tive effect against ketamine-induced apoptosis of neurons by activating the PI3K-Akt signaling pathway.

BACKGROUND:Bone marrow mesenchymal stem cel transplantation for myocardial infarction becomes popularized in recent years, but transplanted cels cannot survive and proliferate under early inflammatory reaction or local ischemia/hypoxia microenvironment, eventualy hampering the therapeutic outcomes.
OBJECTIVE:To investigate the therapeutic effect of PTEN-silenced bone marrow mesenchymal stem cels on acute myocardial infarction.
METHODS:(1) Bone marrow mesenchymal stem cels from Sprague-Dawley rats were randomly assigned to receive no treatment, NCsiRNA transfection using Lipofectamin2000orPTEN siRNA transfection using Lipofectamin2000. Cel growth curves were described using MTT method to detect cel cycle using flow cytometry. (2) Thirty Sprague-Dawley rats were selected to prepare myocardial infarction models that were randomized into three groups (n=10 per group): blank control, negative control and RNAi group. Six hours after modeling, bone marrow mesenchymal stem cels transfected with nothing, NCsiRNA and PTEN siRNA were respectively injected into the infarcted center of the left ventricular anterior wal in these three rat groups. After 4 weeks, al rats were subjected to cardiac function detection using echocardiography, and the survival and proliferation of bone marrow mesenchymal stem cels in the rats were observed by fluorescence microscopy.
RESULTS AND CONCLUSION:Compared with the other two groups, a significant increase in the absorbance values at different culture time, the proportion of cels in S+G2phase, and the number ofbone marrow mesenchymal stem cels in the myocardial tissue was found in the RNAi group (alP< 0.05). Additionaly, the left ventricular ejection fraction and left ventricular shortening fraction were significantly reduced in the RNAi group than the blank control and negative control groups at 4 weeks after cel transplantation (P< 0.05). Bothin vivoandin vitroexperimental findings showed that PTEN silencing could effectively improve cel survival and proliferation in the infarcted myocardium. Moreover, in thein vivoexperiment, an overt improvement in rat’s cardiac function was achieved.

[ Objective ] To supply a technological evidence for the rapid reproduction of Andrographis paniculata (Burm. f.) Nees (APN). [Methods] Different explants from sterile seedling of Andrographis paniculata (Burm. f.) Nees were cultured in vitro to induce the production of fascicular buds. [ Results ] Cotyledon with nodes and half cotyledon with nodes were the most suitable explants for culture, and the budding rate was 100% and over 90% respectively. 6-Benzylaminopurine (BA) in the concentrations of 0.5 ~ 1.0mg/L were beneficial for the reproduction and growth of buds. The age of explants had no correlation with budding. The optimal rooting medium was MT (Murashige and Tucher) medium added with Img/L NAA (naphthaleneacetic acid) or IBA (indolebutyric acid). A highest rooting rate as much as 61.9% was achieved after 10 days of culture. [Conclusion] An effective plant regeneration system has been established for explants of Andrographis paniculata (Burm. f.) Nees.

Objective To analyze the genomic sequences of Cryptococcus neoformans var grubii strains of two genotypes with different virulence and to screen out the virulence-associated genes. Methods A clinical strain (IFM56800) with the strongest virulence and an environmental strain (IFM56731) with the weakest virulence were screened out for whole genome sequencing analysis. The results of sequencing analy-sis were comprehensively analyzed by using the method of comparative genomics. Genetic variations were ex-tensively screened by using the strategies of non-synonymous single nucleotide polymorphisms ( nsSNPs), nonsense SNPs and the insertions or deletions ( InDels) causing frameshift mutations. The filtered genes were sequenced in 20 experimental strains. The whole RNAs were extracted and then the full-length cDNAs were sequenced by using the rapid amplification of 5′ and 3′ cDNA ends (RACE) method. Results By whole genome sequencing, valid data with high coverage (127 times and 111 times) was obtained in both the environmental strain IFM56731 and the clinical strain IFM56800. The data of InDels and SNPs were statisti-cally analyzed, respectively. Six genes were chosen for further analysis based on the strategies of nonsense SNPs and the InDels causing frameshift mutations. The six genes were amplified and sequenced in all of the experimental strains, three of which were further analyzed with cDNA sequencing. Ultimately, the location and structure of CNAG_01032 gene were determined. The predicted nonsense mutation locus was verified to present in the actual mRNA. Conclusion The strategies of nonsense SNPs and the InDels causing frame-shift mutations showed high-efficiency in screening potential virulence-associated genes. The CNAG_01032 gene was screened out as a novel virulence-associated gene.

Objective To detect the serum levels of tumor necrosis factor-alpha (TNF-α) and interferengamma (IFN-γ) in brucellosis patients and to study the Th1 immune response in acute and chronic patients.Method Serum levels of TNF-alpha and IFN-gamma of 110 brucellosis patients,including 58 acute brucellosis patients and 52 chronic brucellosis patients,were measured by enzyme linked immunosorbent assay (ELISA) from 2014 to 2015 in Zhangjiakou Infectious Disease Hospital.Results The serum levels of TNF-alpha and IFN-gamma of 58 acute brucellosis patients were (38.2± 3.6) pg/L and (31.3 ± 3.7) ng/L,respectively;the serum levels of TNF-alpha and IFN-gamma of 52 chronic brucellosis patients were (12.4 ± 2.6) pg/L and (8.8 ± 3.4) ng/L,respectively.The differences were statistically significant between acute and chronic patients (t =43.216,33.809,all P ＜ 0.05).The early cure rate,early base cure rate,improvement rate and inefficiency rate were 36.2％ (21/58),32.7％ (19/58),25.9％ (15/58)and 5.2％ (3/58),respectively in acute patients.Inversely,they were 17.3％ (9/52),13.5％ (7/52),15.4％ (8/52)and 53.8％ (28/52),respectively in chronic patients.The therapeutic effect was better in acutepatients than chronic patients (x2 =4.937,5.657,all P ＜ 0.05).Conclusion It seems that acute brucellosis patients have a higher serum levels of TNF-alpha and IFN-gamma and a better prognosis due to effective Th1 immune response,and chronic brucellosis patients are associated with poor outcome due to deficiency of Th1 immune response.

Aim To investigate the effect of insulin glargine and human insulin on proliferation of a human breast cancer cell line MDA-MB-231 and the role of ERK in the process.Methods MDA-MB-231 cells were incubated with insulin glargine and human insulin at different concentrations and for different time courses.A specific ERK1/2 inhibitor,PD98059,was used either alone or in combination with insulin glargine or human insulin to test the involvement of ERK pathway in cell growth.Cell proliferation was evaluated using cell counting kit-8 reagents.Cell cycle distribution was analyzed by flow cytometry.Results Both insulin glargine and human insulin dose-dependently enhanced MDA-MB-231 cell proliferation at the concentrations from 1 to 100 IU?L-1 after treatment for 96 h.At the concentration of 10 IU?L-1,both drugs promoted cell growth at 48,72,and 96 h.The percentage of S+G2/M cells was significantly increased in both insulin glargine and human insulin treated groups as compared to untreated controls.No significant difference was observed between insulin glargine and human insulin in their effects on cell proliferation and cell cycle distribution.Cell proliferation was significantly inhibited by PD98059.However,in the presence of PD98059,both drugs still promoted cell proliferation significantly as compared to untreated controls.Conclusions Insulin galrgine and human insulin similarly promote proliferation of MDA-MB-231 cells independent of ERK activation.

Objective To explore the advantages of Argus method by comparing the accuracy and timeliness of Argus and artificial methods for measuring femoral head necrosis area in MRI scanning.Methods Totally 17 patients (31 hips) were measured with Argus and artificial methods respectively for the necrosis area, and then the measuring results and time were compared, and the correlation was investigated between the results and the patients' pain degree, along with that between the results and the extent of femoral head collapse.Results The necrosis area ratios determined by Argus and artificial methods were (33.5±4.08)%and (34.6±4.06)%respectively, with no statistical difference between the ratios (P>0.05). The time consumed by artificial method was (21.3 ±3.62)min, significantly longer than (7.89 ±1.03)min by Argus method, with P<0.001. Regression analysis proved that the necrosis areas were positively correlated with the patients' pain degree, and the correlation coefficient by Argus method was 0.807 8, more than 0.740 9 by artificial method. The femoral heads of 11 cases(16 hips) collapsed in the follow-up period, the necrosis areas were positively correlated with the patients collapse level, but the correlation coefficient by Argus method was 0.783 8, more than 0.726 7 by artificial method.Conclusion Argus method gains high accuracy and timeliness when used in MRI scanning of femoral head necrosis area, and thus is worth popularizing clinically.

Objective To discuss the adjustment effects of kidney reinforcing medicine on growth and development and the content of choline acetyl transferase (CHAc) in rats with kidney-deficiency constitution. Methods The method of cats scare rats was used to build composite offspring rat models with deficiency and acquired dystrophy, and then the models were divided into model group, Zuogui Pill group and Yougui Pill group. The rats in blank group came from normal pregnant rats. Baby rats were scared and received gavage at the same time. All administration groups received suspension of Zuigui Pills or Yougui Pills. Blank group and model group received the same amount of normal saline, once a day, for 3 months. Growth and development of rats were observed. Weight and food utilization of rats aged 5–8 weeks in each group were recorded. The content of brain CHAc was detected by ELISA. Results When offspring were 5–8 weeks old, weights of rats in the model group were lower than blank group (P<0.01); while weights of rats in Zuogui Pill group and Yougui Pill group increased significantly (P<0.05) and the food utilization was positively correlated with weight; the CHAc content in the model group decreased significantly compared with blank group (P<0.05, P<0.01); while the CHAc content in Zuogui Pill group and Yougui Pill group increased significantly compared with the model group (P<0.01). Conclusion Kidney reinforcing medicine can improve backward growth and development of rats with kidney-deficiency constitution and adjust CHAc in the brain, so as to promote the learning and memory ability of rats with kidney-deficiency constitution.

Objective To evaluate the sensitivity and specificity of different HbA1C cutoff points for diabetes diagnosis in high risk outpatients in Harbin.Methods A total of 2 122 high risk outpatients(male 1 032 and female 1 090)for diabetes screening in the Fourth affiliated Hospital of Harbin Medical University from April 2013 to February 2015 were included in this study, with the average age of(49.26±13.00)year. Oral glucose tolerance tests(OGTT)were conducted and HbA1C levels were examined in these patients. The sensitivity and specificity of different HbA1C cutoff points were calculated and a receiver operator characteristic(ROC)curve was then built.Results The average level of HbA1C in these subjects was(6.45±1.72)%. The prevalence of diabetes was 41.85%. The area under ROC curve(AUC)was 0.89 with the optimal cutoff point of HbA1C 6.0% and 0.68 for the highest Yonden index. The sensitivity and specificity of HbA1C 6.0% were 84.01% and 83.67% respectively. The sensitivity and specificity of HbA1C 6.5% were 62.84% and 95.92%, respectively. The AUC of HbA1C≥6.5% was 0.732. Conclusion HbA1C works well as the diagnostic standard for diabetes in high risk outpatients of Harbin city. The cutoff point of HbA1C 6.0% is suitable for screening diabetes in high risk population, and HbA1C 6.5% is appropriate for diabetes diagnosis, with high sensitivity and specificity.