Objective To investigate the effect of simulated microgravity on the proliferation and differentiation of the human megakaryocyte cells in vitro. Methods The fourth generation rotating cell culture system (RCCS-4) was used to generate the simulated microgravity environment. The cell viability was assessed by trypan blue staining method. The proliferation of cells was assessed by cell counting method and CCK8 method. The CD41+/CD61+ cells rate and the cells cycle were detected by flow cytometry. The expression levels of thrombopoietin receptor (c-mpl) and transcription factors were detected with RT-PCR. Results After 24, 48, 72 h, culture under simulated microgravity resulted in a significant decrease in the cell number , proliferative activity, cells in the G2/M phase and levels of c-mpl mRNA expression in comparison with that under the normal gravity (P < 0.05). After 48 h and 72 h culture, CD41+/CD61+ cells ratio decreased and RUNX-1 mRNA expression was down-regulated in cells of the group SMG compared with that of the group NG (P < 0.05). Conclusion Microgravity can inhibit the proliferation and differentiation of human megakaryocyte cells in vitro. The mechanism may be that TPO/c-mpl pathway was inhibited by down regulating the expression of c-mpl which transcriptional inhibition lead to.

Objective To observe the protective effect of silybin-phosphatidylcholine compound(SPC) on acute liver damage induced by alpha-naphthylisothiocyanate(ANIT) in mice.Methods Forty mice were randomly divided into 4 groups:normal group,model group,SPCⅠgroup,SPCⅡgroup.Intragastric administration of 0.5% carboxymethylcelluloes-Na(CMC-Na) suspension were given to the normal and model group,and 2 quantities of SPC suspension(150 and 300 mg/kg) were respectively given to SPCⅠ and SPC Ⅱ groups for 1 week.At 12 h after last administration of SPC,all of groups besides normal were given ANIT(dissolved in peanut oil,(60 mg/kg)) by lavage and the normal group just only were given peanute oil by the same volume and same way.After 16 h of ANIT administration,the levels of serum alanine transferase(ALT),aspartate aminotransferase(AST),serum total bilirubin(TB),malondialdehyde(MDA) and superoxide dismutase(SOD) were detected by biochemical method.Results Serum levels of ALT,AST,TB and MDA markedly increased after ANIT was administered via intragastric tube,and the level of serum SOD also decrease.SPC could obviously inhibit the alteration of the activities of serum ALT,AST,TB,MDA and SOD(P

Mycoepoxydiene is a novel antitumor agent extracted from marine lignicolous fungi HLY-2, which is Diaporthe phaseolorum by molecule identification. The medium optimization for mycoepoxydiene by orthogonal design and the comparison of submerged fermentation and solid state fermentation were studied. The rusult is that the maximal yield of the compound is 543mg/L, which is 43 times compared to the customary half-seawater PD medium and 15 times to the best submerged condition. This optimum culture medium included potato 250g/L, seawater 300mL/L, glucose 30g/L, lactose 50g/L, KH_ 2 PO_ 4 0.65mmol/L and (NH_ 4 )_ 2 SO_ 4 1g/L in the solid state condition. Differentiation analysis between submerged and solid state fermentation, and antitumor activity of these ferment products were also studied. The antitumor activity of products of the optimum medium approached the pure compound.

Objective : To evaluate the effectiveness of interventions for human papillomavirus ( HPV ) vaccine hesitancy among female university students based on the precaution adoption process model (PAPM), so as to provide the evidence for improving the coverage of HPV vaccine in this population. Methods : HPV vaccine hesitant female students were selected using a cluster sampling method from Zhejiang Chinese Medical University, and randomly assigned to the PAPM group and control group. Students in the PAPM group received PAPM-based interventions for HPV vaccine hesitancy, while students in the control group were given routine interventions. The HPV-related knowledge, HPV vaccine-related knowledge and HPV vaccine hesitancy scores were collected from both groups prior to interventions ( T0 ), 0 ( T1 ), 1 ( T2 ) and 3 months post-interventions ( T3 ), and the effectiveness of interventions was evaluated using analysis of variance for repeated measures. Results : There were 147 students in the PAPM group and 141 students in the control group. In the PAPM group, 36.73% of the students majored in medical sciences, and 48.23% were freshmen; in the control group, 39.72% majored in medical sciences, and 50.35% were freshmen. The mean scores of HPV- and HPV vaccine-related knowledge were significantly greater in the PAPM group than in the control group, respectively ( T1, 5.29 vs. 4.91; T2, 4.27 vs. 4.22; T3, 4.22 vs. 4.04; P<0.05 ); however, no significant differences were detected in the HPV vaccine hesitancy scores between the two groups, respectively ( T1, 2.98 vs. 2.95; T2, 3.07 vs. 3.07; T3, 3.08 vs. 2.97; P>0.05 ). The mean scores of the confidence dimension ( T1, 3.37 vs. 3.23; T2, 3.48 vs. 3.40; T3, 3.38 vs. 3.25 ) and the dimension of influence by others ( T1, 3.44 vs. 3.33; T2, 3.42 vs. 3.37; T3, 3.46 vs. 3.27 ) were significantly greater in the PAPM group than in the control group (P<0.05), while the mean scores of the complacency dimension were significantly lower in the PAPM group than in the control group ( T1, 1.98 vs. 2.03; T2, 2.06 vs. 2.20; T3, 2.18 vs. 2.15; P<0.05 ). Conclusions PAPM-based interventions for HPV vaccine hesitancy may effectively improve the awareness of HPV and HPV vaccines, reduce complacency, and enhance the influence by others among female university students.

OBJECTIVE: To study the efficacy and safety of Yiqi Tongmai Oral Liquid (YQTM), a traditional compound Chinese herbal medicine, in treating angina pectoris in patients with coronary heart disease. METHODS: A multicentric, randomized, double blinded and paralleled controlled trial was conducted on 110 patients in trial group treated with YQTM, and 109 patients in control group treated with Shuxin Oral Liquid (SX). Cure and effective rates in both groups were evaluated. Frequency and duration of angina attack were counted and measured. Coronary angiography (CAG), electrocardiogram (ECG) and flat exercise test were taken in both groups. Blood lipid indexes, such as cholesterol (CH), triglyceride (TG), low-density lipoprotein (LDL), high density lipoprotein (HDL), were determined at pre- and post-treatment. The hemodynamic indexes, such as whole blood viscosity (J2), high-shear reduced viscosity (Eh), low-shear reduced viscosity (Ei), red cell aggregation index (Lb), red cell rigidity index (Rh), fibrinogen (Fb), blood sedimentation rate (BSR) and hematocrit (HCT), were determined at pre-and post-treatment. The indicated scores of symptoms and signs of traditional Chinese medicine (TCM) pattern, such as chest pain, chest constriction, breath shortness, palpitation, fatigue, dim complexion, spontaneous perspiration and tongue proper, tongue coating were evaluated in week 0, 1, 2, 3, 4 during the treatment course. The safety indexes, such as body temperature, pulse, respiration and blood pressure were observed. Routine tests of blood, urine and stool, hepatic function test and renal function test were taken at pre- and post-treatment. RESULTS: There was no significant difference between the total effective rate of the trial group and that of the control group, which were 91.82% and 85.32%, respectively (P>0.05). Trial groups percentile of cure rate is significantly higher than that of the control group (P<0.01). The frequency and duration of angina attack, the positive ratio of CAG and flat exercise test of both groups were lowered, while the effect of the trial group on frequency and duration of angina attack was better. No significant difference was found in ECG features between the two groups (P>0.05). The levels of CH, TG and LDL of both groups were lowered significantly (P<0.05). The effect of lowering CH, TG and LDL of the trial group was stronger than that of the control group (P<0.05). The hemodynamic indexes, such as J2, Eh, Ei, Lb, Rh, Fb, BSR and HCT were improved significantly in both groups (P<0.05). The improvements of J2, Eh, Ei, Lb, Rh, Fb and SR in the trial group were greater than those of control group (P<0.05). The TCM symptoms and signs, such as chest pain, chest constriction, breath shortness, palpitation, fatigue, dim complexion, spontaneous perspiration were improved significantly in both groups (P<0.05). The improvements of chest constriction, palpitation, fatigue and spontaneous perspiration in the trial group were greater than those of the control group (P<0.05). The total indicated score of TCM symptoms and signs was lowered more significantly than that of the control group (P<0.01). No significant changes were found at pre- and post-treatment in safety indexes, such as routine tests for blood, urine and stool, hepatic function test and renal function test. There was no significant difference in safety features of both groups (P>0.05). CONCLUSION: Yiqi Tongmai Oral Liquid bears good therapeutic effect on angina pectoris without adverse reaction, and is superior to Shuxin Oral Liquid. Yiqi Tongmai Oral Liquid is a new effective and safe medicine for the treatment of angina pectoris in patients with coronary heart disease.

Objective Pemetrexed and β-elemene can inhibit the growth of tumor cells .This study was to investigate the effect of pemetrexed combined with β-elemene on the proliferation and apoptosis of cervical cancer HeLa cells. Methods Cervical cancer HeLa cells were treated with pemetrexed at the concentrations of 38, 76, 152, 228, and 304μg/mL, and at 24 and 48 hours of treatment subjected to MTT for detection of their proliferation .The experiment included four groups , with the cells treated with β-elemene ( 125μg/mL) , pemetrexed ( 76 μg/mL ) , β-elemene ( 125 μg/mL ) +pemetrexed (76μg/mL), and nothing (blank control) for 24 hours, followed by determination of their proliferation and apoptosis by MTT and flow cytometry, respectively. Results Pemetrexed at 38, 76, 152 and 228μg/mL inhibited the proliferation of the HeLa cells in a concentration-dependent manner, with the inhibition rates of (7.24 ±3.78), (7.94 ±4.37), (11.10 ±2.86) and (15.88 ± 3.38)%at 24 hours, and (16.69 ±0.95), (22.54 ±1.53), (24.48 ±0.92) and (25.54 ±3.61)%at 48 hours, both with statis-tically significant differences between any two groups (P<0.05).Significant differences were also found in the proliferation rate of the same concentration of pemetrexed at the two time points (P<0.05).The combination of pemetrexed and β-elemene showed an inhibi-tion rate of (49.95 ±5.76)%at 24 hours, remarkably higher than (24.36 ±5.59)%in theβ-elemene group and (10.69 ±1.37)%in the pemetrexed group (P<0.01). Conclusion Pemetrexed combined with β-elemene can significantly inhibit the proliferation and synergistically accelerate the apoptosis of HeLa cells .

OBJECTIVETo investigate polymorphisms in the coding exons and their splicing areas of caspase10 gene (CASP10) in Chinese of Han nationality.

METHODSPolymerase chain reaction (PCR), denaturing high-performance liquid chromatography (DHPLC), direct sequencing and clonal sequencing were used to analyze the exon 9 and its flanking sequences.

RESULTSIn 70 blood samples of Chinese subjects researched the known single nucleotide polymorphisms (SNPs) in the exon 9 of CASP10 gene published in dbSNP of NCBI were not identified. We found a short sequence with more than ten continuous and repeated T nucleotides within the intron 8 near the exon 9 and the length of repeated sequence nucleotides T was different in samples. Clonal sequencing analysis indicated that it was a microsatellite of single nucleotide-repeated sequence. The recurrence and specificity of same result was further confirmed by PCR with high fidelity polymerase and sequencing. Furthermore all of 70 blood samples detected by DHPLC showed heterozygous profiles. We named it as IVS8-13(T)n temperately.

CONCLUSIONOur research suggested that the site be a microsatellite of single nucleotide-repeated sequence. Meanwhile, our data further showed that it was quite different from that by querying SNP database of non-Chinese population in NCBI, in the same region a SNP of type of insertion deletion existed. It could be inferred that the difference between the populations might be associated with the genetic characteristics of ethnics. The significance of the microsatellite in CASP10 gene in Chinese of Han nationality remains to be elucidated.

Asian Continental Ancestry Group ; genetics ; Base Sequence ; Caspase 10 ; Caspases ; genetics ; China ; ethnology ; Ethnic Groups ; Genetics, Population ; Humans ; Introns ; genetics ; Microsatellite Repeats ; genetics ; Molecular Sequence Data ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA

In order to research developmental competence of transgenic somatic cell by serial nuclear transplantation, goat cloned embryos were compared with recloned embryos in ability of in vitro development. Fetal fibroblasts including human finger-domain lacking t-PA gene was microinjected into cytoplasm of the MII oocytes. Goat embryos (G0) were cloned by this procedure. A single blastomere from 16 - 64-cell goat cloned embryos (G0) was microinjected into Intracytoplasm of the MII oocytes. Goat embryos (G) were cloned by this procedure. Goat embryos (G2, G3) were recloned by using 16 - 64-cell recloned embryos. The developmental time of donor embryo affected the developmental rate of recloned embryos (G1, G2). The results show: the cleavage rate of cloned embryos (G0) (76.45% +/- 1.17%) was no difference significantly with recloned embryos (G1 G2 G3) (72.18% +/- 1.97%, 76.05% +/- 2.38%, 75.99% +/- 2.84%); the developmental rate of morulae and blastocysts of cloned embryos (47.20% +/- 2.93%, 11.00% +/- 1.42%) were higher than these of recloned embryos(34.99% +/- 2.66%, 28.23% +/- 2.00%, 23.34% +/- 1.99%) (3.87% +/- 0.67%, 2.08% +/- 1.66%, 0); the morulae rate(29.57% +/- 1.53%, 24.43% +/- 1.87%) and blastocysts rate(1.96% +/- 1.31%, 2.01% +/- 1.34%) of recloned embryos (G1 G2) from 16-cell recloned embryos were lower than those(34.32% +/- 1.31%, 29.76% +/- 1.66% and 3.86% +/- 1.03%, 3.48% +/- 0.34% )from 32 - 64-cell recloned embryos (P > 0.05). In conclusion, nuclear transfer embryos should not were recloned mostly; and the embryos recloned by using 32 - 64-cell embryos achieved higher developmental ability compared with using 16-cell embryos.
Animals ; Animals, Genetically Modified ; Blastomeres ; cytology ; physiology ; Cloning, Organism ; methods ; veterinary ; Embryo Culture Techniques ; Embryo, Mammalian ; cytology ; Female ; Fibroblasts ; cytology ; Gene Deletion ; Goats ; Humans ; Nuclear Transfer Techniques ; veterinary ; Oocytes ; cytology ; physiology ; Pregnancy ; RING Finger Domains ; genetics ; physiology ; Tissue Plasminogen Activator ; genetics ; metabolism

OBJECTIVETo investigate the relationship between the frequency of BCL-2/IgH rearrangement in peripheral blood cells of healthy Chinese individuals of Han nationality located in Zhejiang area and the low incidence of follicular lymphoma (FL).

METHODSNested-PCR and direct DNA sequencing were used to detect the Bcl-2/IgH rearrangement in peripheral blood cells of 196 healthy individuals. DNA sequences involved were then searched and aligned in NCBI database to confine the broken points in major breakpoint region and the IgH segments involved.

RESULTSFirst, in this sample the frequency of BCL-2/IgH translocation in Chinese individuals of Han nationality located in Zhejiang area is 9.66%, being much lower than that in North America and Europe countries. Second, the breakpoints tend to fall into 3 clusters: 3055, 3116 and 3165 bp. Usage of J6 segment is most common. Third, There are different subclones of BCL-2/IgH rearrangements in the same individual.

CONCLUSIONThe low frequency of BCL-2/IgH translocation in healthy Chinese individuals of Han nationality located in Zhejiang area may be one of the reasons for the difference in the incidence of FL between China and Western countries.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Chromosomes, Human, Pair 14 ; Chromosomes, Human, Pair 18 ; Female ; Gene Rearrangement ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Lymphoma, Follicular ; ethnology ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; Polymerase Chain Reaction ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Translocation, Genetic ; Young Adult

Objective To summarize the implementation process of Health Project for Residents and evaluate the effect, so as to provide reference for the successive advance development of the project. Methods Field investigation combined with current data review was carried out and 2 498 residents were sampled by multi-stage cluster sampling method. Results With the general goal of Healthy Anting, health promotion programs named “12345”( i.e.one investigation, two carriers, three platforms, four programs and five approaches) had been conducted from 2012 to 2015, which promoted health into fami-lies, communities and companies greatly.The project awareness rate of residents was 63.6%, which was 4.0% higher than that in 2012.91.7% of residents were totally satisfied with the project.The life expectancy was 83.22 in 2015, which was 0.52 years higher than that in 2012.Infectious diseases were controlled in low level and the incidence of categories A and B was 101.78 per hundred thousand.The current prevalence of cancer was 2.43%, which was higher than that in 2012 (1.83%).The cigarette smoking rate was 22.30%, lower than that in 2012(25.56%).The proportion of residents who attended physical exercise in spare time was 45.5%, higher than that in 2012 ( 30.6%).Meanwhile, the prevalence of residents in two weeks was 6.3%, lower than that in 2012 (10.3%).Additionally, the proportion of visiting community center as their first choice was 40 .20%, which was higher than that in 2012 ( 38 .1%) . Conclusion Remarkable achievements had been made and the Health Project for Residents should be carried out successively on the basis of local reality as well as the health related policies, so as to satisfy the health demand and total health level of residents.