Cardiac muscles of mouse embryos, newborn and suckling mice were used for histological and histochemical studies. This paper presents the observations on the morphogenesis and ultrastructure of cardiac myocytes under electron microscope and on their reactions of nucleic acids (DNA and RNA), glycogen, lipids, succinodehydrogenase (SDH), alkaline and acid phosphatases (AKP and AcP), glucose-6-phosphatase (G-6-Pase) and adenosine triphosphatase (ATPase) under light microscope.The heart of mouse embryo before 12-day, contained numerous polygonal or star-shaped myoblasts which had not yet acquired myofibrils but a few myofilaments. As the myoblast developed, the number of myofilaments increased in number and formed myofibrils, then the cells became myocytes. By the end of embryonic period, all the special elements of myocyte were basically constituted. The myocytes of embryos were rich in RNA granules, and their DNA was deeply stained. Flourishing mitosis appeared only in the early embryonic phase. RNA of adult myocytes was much less than that of embryos. From the early phase of embryos myocytes were full of glycogen but short of lipid droplets. From the day it was born, glycogen decreased apparently but lipid droplets increased rapidly.The reaction of SDH steadily increased in intensity from its early phase to late one. After birth it became more intensive. G-6-pase first appeared in the myocytes of 14 day's embryos. In the fetal period it showed moderate positive reaction, but in the myocytes of suckling muose it appeared negative. The enzyme showed positive reaction again at the age of 2 weeks. The ATPase reaction was found to be weak in the fetal specimens, only appeared in the endothelium of the capillaries. After birth it gradually became intensive and from the 2nd week positive reaction was obvious in adult, it was very vigorous.The above observations showed that the embryonic development and differentiation were gradually completed. Histological and histochemical features of each developmental period showed their individualities, which confirmed the evidence that the cardiac muscle developed not only successively but also by stages, and approached adult's level at 2nd week end after birth.

Objective To investigate the effect of atovastatin combined with trimetazidine on cardiac function and inflammatory cytokines in patients with chronic heart failure(CHF). Methods Ninety-seven patients with CHF were randomly divided into three groups:control group treated with conventional therapy,atovastatin group treated with atovastatin based on conventional therapy, and combined treatment group treated with atovastatin and trimetazidine based on conventional therapy. The parameters of cardiac function, including left ventricular ejection fraction ( LVEF) and left ventficular end-diastolic diameter (LVEDD) ,and the plasma levels of CRP,brain natriuretic peptide(BNP) ,and TNF-α were detected in all patients of each group before and after treatment. Results The plasma levels of CRP,BNP and TNF-α in patients with cardiac function Ⅳ were significantly higher than in those with cardiac function ≤ Ⅲ (P<0. 05). There was significant difference in plasma levels of CRP, BNP,TNF-α,LVEF and LVEDD before and after treatment in all groups(P<0. 05 or P<0. 01). As compared with control group after treatment,the plasma levels of CRP,BNP and TNF-α after treatment in both atovastatin group and combined treatment group were decreased markedly(P<0. 05),so did LVEF, LVEDD and 6-min walk test(P<0. 05). There was no significant difference in changes of CRP,BNP and TNF-α plasma levels,LVEF and LVEDD between atovastatin group and combined group after treatment (P>0. 05),but 6-min walk test lengthened statistically in combined treatment group (P<0. 05). Conclusion Atovastatin based on conventional therapy may play a role in anti-inflammation by lowering the plasma levels of CRP, BNP and TNF-α in patients with CHF, thereby improving cardiac function. Atovastatin combined with tremetazidine could reduce the cardiac muscular oxygen consumption and raise the excise endurance in patients with CHF.

Objective To study the expression of APE/Ref-1 in gastric cancer. Methods Detection of APE/Ref-1 protein was performed with gastric cancer tissue microarray (TMA) by IHC in gastric cancer, non-neoplastic mucosa and lymph node with metastasis. Results The positive rates of APE/Ref-1 in cytoblast in non-neoplastic mucosa, tumor tissue, and metastatic lymph nodes were 96.6%, 97.1% and 98.9%, respectively, while the positive rates in cytoplasm were 71.8%, 21.4% and 10.0%, respectively and in both cytoblast and cytoplasm were 71.4%, 21.4% and 10.0%, respectively. Compared to non-neoplastic mucosa, tumor lesion had lower APE/Ref-1 expression in cytoblast and cytoplasm. In 35.9% of patients it was decreased slightly and in 11.2% of them it was decreased markedly in cytoblast (P

To localize and characterize the hypothetical protein CT358 in the chlamydial infected cells.CT358 gene from the Chlamydia trachomatis(C.trachomatis) serovar D genome was amplified and cloned into the pGEX and pDSRedC1 vectors.The recombinant plasmid pGEX-CT358 was constructed and expressed as GST fusion proteins.The GST-CT358 fusion protein was used to immunize mice to raise the antibodies,which specifically recognized CT358 without cross-reacting with other unrelated proteins.The antibodies were then used to localize the endogenous CT358 protein and determine the expression pattern in Chlamydial infected cells using an indirect immunofluorescence assay(IFA).Meanwhile,pDSRedC1-CT358 recombinant plasmid was transfected to HeLa cells to evaluate the effect of CT358 expression on the subsequent chlamydial infection.The hypothetical protein CT358 was identified in the inclusion membrane of C.trachomatis-infected cells for the first time,and it was detected as early as 12 h after C.trachomatis infection and remained in the inclusion membrane throughout the rest of the infection cycle.Cytosolic expression of CT358 via a transgene failed to affect the subsequent chlamydial infection.These observations together have demonstrated that CT358 is a newly identified chlamydial inclusion membrane protein,giving the potentially importance for further understanding the mechanisms of chlamydial intracellular parasitism.

To localize and characterize the hypothetical protein CT358 in the chlamydial infected cells. CT358 gene from the Chlamydia trachomatis (C. trachomatis) serovar D genome was amplified and cloned into the pGEX and pDSRedCI vectors. The recombinant plasmid pGEX-CT358 was constructed and expressed as GST fusion proteins. The GST-CT358 fusion protein was used to immunize mice to raise the antibodies, which specifically recognized CT358 without eross-reacting with other unrelated proteins. The antibodies were then used to localize the endogenous CT358 protein and determine the expression pattern in Chlamydial infected cells using an indirect immunofluorescence assay (IFA). Meanwhile, pDSRedC 1-CT358 recombinant plasmid was transfected to HeLa cells to evaluate the effect of CT358 expression on the subsequent chlamydial infection. The hypothetical protein CT358 was identified in the inclusion membrane of C. trachomatis-infected cells for the first time,and it was detected as early as 12 h after C. trachomatis infection and remained in the inclusion membrane throughout the rest of the infection cycle. Cytosolic expression of CT358 via a transgene failed to affect the subsequent ehlamydial infection. These observations together have demonstrated that CT358 is a newly identified chlamydial inclusion membrane protein, giving the potentially importance for further understanding the mechanisms of chlamydial intracellular parasitism.

【Objective】To explore the role of cell cycle regulators in nasopharyngeal carcinoma (NPC) relapse.【Method】To assay p53,MDM2,p21ras and p21WAF1 proteins by LsAB immunohistochemical technique in 69 cases of primary and relapsing NPC tissues.【Results】As compared with primary NPC,the expression rate of p53 or MDM2 protein in relapsing NPC was similar (78% to 80%,84% to 83%),and the expression rate of p21ras or p21WAF1 protein in relapsing NPC was obviously descended (73% to 93%,52% to 84% );the high-expression rate of p53 protein in relapsing NPC was similar (42% to 51%),the high-expression rate of MDM2 protein in relapsing NPC was obviously risen (57% to 32%),and the high-expression rate of p21ras or p21WAF1 protein in relapsing NPC was obviously descended (16% to 65%,17% to 46%).Among of them,the significant rise of MDM2 protein expression level in relapsing NPC mainly occurred in the patients of group 2 which relapsing-interval was shorter than 34 months,P<0.05;the significant descent of p21ras or p21WAF1 protein expression level in relapsing NPC occurred in the patients of group 2 and group 1 which relapsing-interval was equal to or longer than 34 months,P<0.02,respectively.【Conclusions】The overexpression of p53 and MDM2 proteins and the low or negative expression of p21WAF1 protein after clinical cure might still play an important role in NPC relapse,the obvious rise of MDM2 protein level and the obvious descent of p21WAF1 protein level might further accelarate the process of NPC relapse.

Objective To evaluate the value of treating chotestatic hepatitis with tolynicate and naphthylacetia acid,and Dansen Root rejected fluid.Methods All patients were divided into Control Group treated with Potassium magnessium aspartape,Glucurolactone etc,and Treating Group treated with tolynicate and naphthylacetia acid,and Dansen Root re- jected fluid;The change of clinical symptom,jaundice decreasing and recovery of hepatic function were observed respec- tively.Results The change of clinical symptom,jaundice decreasing and recovery of hepatic function of Treating Group were better,and there are significant difference (P

microRNAs (miRNAs) are an extensive class of -22-nucleotide (nt) endogenous noncoding RNAs regulating life activities ofmetazoans through binding to 3'-untranslated regions (3'-UTRs) of their target genes. This work aimed to identify yan gene in the silkworm, reveal its expression profile and confirm if it is one target of bmo-miR-7 and, as such, have potential for contributing to better understanding of the molecular mechanisms involved in the metamorphosis of silkworm. Based on homolog searching and PCR amplification, we cloned the coding sequence (CDS) of Bmyan, which encodes 476 amino acid residues and contains SAM-PNT and ETs domains. Quantitative PCR (q-PCR), RT-PCR and microarray data revealed high expression of Bmyan in the head, body wall and ovary of day-3 fifth instar larval silkworm, low or no expression in other tissues. It was lowly expressed in the early larval stages, but highly expressed from late spinning to day 4 pupa. The 3'-UTR of Bmyan was obtained by rapid-amplification of cDNA ends (3'RACE) and predicted to contain two potential recognition sites of bmo-miR-7. The luciferase reporter vector containing the 3'-UTR of Bmyan was constructed and co-transfected into BmE cell line with the mimic of bmo-miR-7 and the decreased relative activity of luciferase showed that Bmyan is one target of bmo-miR-7. This work helps further functional analysis of bmo-miR-7 and Bmyan in the silkworm.
3' Untranslated Regions ; Animals ; Bombyx ; genetics ; Cell Line ; Cloning, Molecular ; Female ; Genetic Vectors ; Insect Proteins ; genetics ; Larva ; Metamorphosis, Biological ; MicroRNAs ; genetics ; Pupa

Objective To investigate whether there existed a healthy obese subtype.Methods A total of 116 healthy subjects were recruited.They were divided into 3 groups according to BMI and metabolic disorders:40 cases of normal weight and metabolic normality (NMN),36 cases of obesity and metabolic normality (OMN) and 40 cases of obesity and metabolic abnormality (OMA).Anthropometic parameters as height,weight,waist circumference,hip circumference and blood pressure was recorded.Blood glucose,lipids,insulin,high-sensitivity C-reactive protein (hs-CRP) was detected.Body fat distribution was detected by dual-energy X-ray absorptiometry (DXA).Serum von Willebrand factor (vWF),a marker of endothelial dysfunction,were detected by ELISA.Results Both serum vWF levels in OMN group [(733.6±86.2)U/L] and OMA group[(809.2 ±46.3)U/L] are higher than that in NMN group [(466.9 ±65.3)U/L,P ＜0.05] with serum vWF level in OMA group is higher than in OMN group (P ＜ 0.05).Among android fat mass percentage (AFM％),BMI,waist height ratio,waist circumference,hs-CRP,weight,hip circumference and trunk fat mass,AFM％,BMI and hs-CRP are main influencing factors of vWF.Conclusions Endothelial dysfunction existed in obese adults regardless of their metabolic status.There is no healthy obese subtype.AFM％,BMI and hs-CRP are the main influencing factors of endothelial dysfunction.

Aim To study the therapeutic effect of CpG-ODN, an agonist of Toll-like receptor 9 (TLR9), on hypoxic/ischemic encephapathy in neonatal rats and investigate the mechanisms.Methods Fifty healthy 7-day-old neonatal Wistar rats (in either gender, weighing 12~17g) were randomly divided into sham operation group, HIBD group, and CpG-ODN low group(0.35 mL·kg-1), CpG-ODN middle group(1.40 mL·kg-1), CpG-ODN high group(5.60 mL·kg-1).The neurological function was scored after 48h operation;ten rats of each group was executed respectively and brains tissue was taken;HE staining was used to observe the brain pathological changes.Western blot assay was used to detect the expressions of TLR9 and phosphor-p38 mitogen-activated protein kinases(p-p38 MAPK), and enzyme linked immunosorbent assay (ELISA) method was adopted to detect TNF-α expression.Results The CpG-ODN low, middle group were improved in impairment significantly compared with the HIBD group, and the brain pathological change was lessened, while the CpG-ODN high group was impaired significantly compared with the HIBD group (P<0.05), and brain pathological change was sharpened.Western blot showed the up-regulation in TLR9 and p-p38 MAPK and a significant increase of the expression of TNF-α in the brain tissue in CpG-ODN group with statistical difference in HIBD group and sham operation group(P<0.05).Conclusions The neuro-behavioral score and nervous system function can be improved and the hypoxic/ischemic brain damage can be reduced in neonatal rats in the CpG-ODN low, middle group.The protective mechanisms may be suitably via activating p38 MAPK signaling pathway to promote p38 MAPK phosphory1ation and up-regulation of the expression of TNF-α in the brain tissue of rats.