BACKGROUND:The pathogenesis of avascular necrosis of the femoral head is still uncertain,so the treatment is not favorable.OBJECTIVE:To study the feasibility and mechanism of intramedullary injection with salvia miltiorrhiza for preventing of steroid-induced necrosis of the femoral head.DESIGN,TIME AND SETTING:Randomized controlled animal trial was performed at Zunyi Medical College between April 2005 and May 2007.MATERIALS:Thirty 6-month-old healthy rabbits,half amount of male and female,weighing(2.5?0.25) g,were randomly divided into three groups(n=10):control,model,and prevention groups.METHODS:Normal saline was injected into the control group.Prednisolone was injected into model group,7.5 mg/kg,two times per week for 8 weeks.Glucocorticoid-induced animal model in the prevention group was intramedullary injected with salvia miltiorrhiza,0.3-0.5 cm below the third trochanter of the femur,0.4 mL/kg,twice a week for 8 weeks.MAIN OUTCOME MEASURES:Blood lipids level,serum calcium and phosphorus were measured;X-ray,emission computerized tomographic(ECT) and histopathology were performed.RESULTS:The serum calcium and phosphorus in model group were remarkably reduced.X-ray showed inhomogeneous density of the femoral head,with irregular radiolucent area,obscure structures of bone-trabecula,but intact femoral head and normal joint space.The blood flow and blood pool showed decreasing radioactive distribution of the femoral head,and local concentration of nuclide in the delayed phase in the model group.Histopatholgical observation suggested that the cortical bone was thinned,with thin bone-trabecula and irregular arrangement,necrosed bone marrow and pimelosis and increased bone lacuna.The calcium-phosphorus product of prevent group was increased;X-ray showed normal femoral head appearance and normal bone density except unclear bone-trabecula.ECT showed the dynamic and static images of the prevention group were similar as normal imaging of rabbits.In addition,histopatholgical observation suggested the rate of empty lacuna was decreased,and bone trabecula minimally thinned with ordered arrangement.CONCLUSION:Salvia miltiorrhiza injection plays a good effective role in the prevention of steroid-induced avascular necrosis of femoral head through improving hemorheology and reducing rate of empty lacuna.

Donguibogam was introduced from traditional Chinese medicine (TCM) instead of the local representa-tive work of the Democratic People's Republic of Korea (DPRK). It is also the concrete manifestation of theories of different schools of doctors according to Dr. Xu Jun's medical thoughts. In this paper, from Dr. Xu's advocacy to Taoism and the concept of aftercare spirit, the exploration was made on health cultivation for different body constitu-tions. According to the health cultivation effects of the medicinals and edible food, the focus was made on the sys-tematic collation and induction of more than 1 400 kinds of herbs contained in the soup liquid from papers in three volumes. The medicinals were divided into eight categories, which were benefiting qi, nourishing yin, tonifying yang, clearing heat and promoting dieresis, drying dampness and resolving phlegm, promoting blood circulation for remov-ing blood stasis, dispersing stagnated liver qi, nourishing blood for hemostasis. Donguibogam specified the health cultivation concept from the correspondence between the body constitution and the medicinal performance. It has a certain guiding significance to the current health cultivation.

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) can grow in host myocardium, differentiate under myocardial condition, improve cardiac function. However, biological characteristics of BMSC differentiation are still unclear presently.OBJECTIVE: To study the expression and electrophysiological characteristics of BMSC/n vitro connexin-43 following 5-azacitidine (5-aza) treatment.DESIGN, TIME AND SETTING: The cytological in vitro controlled study was perormed at the Heart Center, Hebei Provincial People's Hospital from July 2007 to February 2009.MATERIALS: A total of 24 male pigs aged 2 months were purchased from Exparimental Animal Center, Hebei Medical University.METHODS: Bilateral femoral bone marrow was obtained from pigs under sterile condition. BMSCs were harvested by Percoll density gradient in vitro. At passage 2, BMSCs were treated with 10 μmol/L 5-aza, and incubated in DMeM without inductor 24 hours later. Indices were measured I, 2, 3 weeks following induction. A control group was set up, which was not treated with 5-azacitidine. Following bone marrow extraction, experimental pigs were anesthetized to obtain ventricular muscle. Normal ventricular muscle cells were isolated and cultured by tissue block enzyme digestion method.MAIN OUTCOME MEASURES: Expression of connexin-43 was measured by immunohistochemical staining (ABC method). Ito current density and action potential were determined by patch clamp technique.RESULTS: At 1 and 2 weeks following 5-aza induction, some BMSCs were positive for connexin-43, with the presence of brown particles surrounding nuclei. At 3 weeks, positive rate of connexin-43 was 95%. The area with large cell density was presented with similar structure to normal myocardium. At +80 mV, compared with normal myocardial cells, Ito current density was significantly reduced in BMSCs following 1 and 2 weeks and in the control group (P < 0.05). Ito current density was significantly increased to a normal levels in BMSCs 3 weeks following induction (P > 0.05). No action potential was detected in BMSCs following 1 and 2 weeks of 5-aza, and action potential could be determined 3 weeks following induction, which was identical to normal myocardial cells.CONCLUSION: Through induced by 5-aza for three weeks, BMSCs have the similar expression of connexin-43 and electrophysiological characteristics as normal myocardium.

AIM:pUL23,the product of human cytomegalovirus (HCMV) gene UL23 was identified as one of tegument proteins. The aim of this research is to investigate the function of pUL23 during HCMV life cycle.METHODS:GAL4 yeast two-hybrid assay was performed to screen the human fetal kidney cDNA library to obtain host cell protein molecules which interact with pUL23 of HCMV. Then the GST-pulldown experiment was applied to confirm the protein interactions identified by yeast two-hybrid.RESULTS:ATPase inhibitory factor 1 (ATIF1) was selected from host cells using yeast two-hybrid assay. GST-pulldown experiments in vitro further confirmed the interaction between ATIF1 and pUL23.CONCLUSION:pUL23 of HCMV can interact with ATIF1 in host cell,which may provide the evidence for understanding the function of pUL23 in the life cycle of HCMV.

Objective To evaluate the immune effects of bivalent inactivated rotavirus vaccine (IRV) and investigate the viability of development of bivalent IRV.Methods Firstly,bivalent IRV was prepared by mixing G1 IRV and G3 IRV with equal amount,G1 IRV and G3 IRV as monovalent control,PBS as negative control.Secondly,those vaccines were vaccinated to the mice by intramuscular injection.Then,to evaluate the immune effects of bivalent IRV,the levels of serum or fecal rotavirus specific IgG and IgA were assessed by ELISA,the levels of serum neutralized antibody were measured by microneutralization assay,the number of IFN-γ or IL-4 secreting cells were analyzed by ELISPOT assay.Results Compared to negative control group,bivalent IRV induced the higher levels of serum and fecal G1 and G3 rotavirus specific antibody.It was found that there were no significant differences for the levels of serum IgG and IgA,fecal IgG and IgA,serum neutralized antibody between induced by bivalent IRV and induced by G1 type monovalent vaccines ; but there were significantly increase for the levels of serum IgG (t =2.691,P＜0.05) and serum neutralized antibody (t =2.561,P＜0.05) between induced by bivalent IRV and induced by G3 monovalent vaccines,there were no significant differences for other antibodies between induced by bivalent IRV and induced by G3 monovalent vaccines.At the same time,compared to negative control group,bivalent IRV induced significantly increase in the number of IFN-γ or IL-4 secreting cells in spleen lymphocytes.It was found that there were no significant differences for the number of IFN-γ or IL-4 secreting cells stimulated by G1 rotavirus between bivalent IRV and G1 monovalent vaccines; but there were significantly increase for the number of IL-4 secreting cells (t =2.327,P＜0.05) stimulated by G3 rotavirus between bivalent IRV and G3 monovalent vaccines,there were no significant differences for the number of IFN-γ secreting cells stimulated by G3 rotavirus between bivalent IRV and G3 type monovalent vaccines.Conclusion The bivalent IRV can induce effective immune response,in which there were no inhibitory interference between the components of bivalent IRV,which provided the experimental basis for the development of bivalent IRV.

Objective To investigate the influence of two different dose of atorvastatin on the prognosis and endothelial function in post-PCI acute coronary syndrom patients.Methods 92 post-PCI ACS patients were randomly divided into two groups,atorvastatin 20mg and atorvastatin 10 mg group.In each group the patients were treated with atorvasta- tin 20mg or 10mg respectively.After one month we add or decrease the dose of atorvastatin according to the blood lipid level.After 12 month the blood lipid level、FMD、NO、ET、NOS、UAP、AMI were compared between two groups. Results The clinical setting have no significant association between two groups before treating,After treated 1 and 12 month the TC,LDL-C level were significantly decreased as compared with the base level before treating in each group. After treated 1 month,in atorvastatin 20 mg group the TC,LDL-C level were significantly decreased and NO、NO/ET level were significantly higher than those in atorvastatin 10 mg group.During 12 month follow up the incidence of angina pectoris onset and rehospitalization were significantly higher in atorvastatin 10 mg group(P

BACKGROUND:Bone marrow stem cell transplantation can improve heart function and prevent ventricle remodeling.At present,the adult bone marrow stem cells used for transplantation primarily included bone marrow mononuclear cells (BM-MNCs) and mesenchymal stem cells (MSCs),and endothelial progenitor cells.The curative effects and precise mechanisms of transplantation of various bone marrow stem cells remain unknown.OBJECTIVE:To compare the effects of transplantation of autologous BM-MNCs and MSCs via the coronary artery on ventricle remodeling subsequent to acute myocardial infarction (AMI). DESIGN,TIME AND SETTING:Randomized controlled animal experiment performed at the Center for Clinical Research,Hebei Provincial People's Hospital,Electron Microscope Room,Hebei Medical University between March 2005 and December 2006.MATERIALS:Thirty-six male Jizhong pigs,were randomly divided into 4 groups:control group (n = 6),infarct model group (n = 10),BM-MNC group (n = 10),and MSC group (n = 10).METHODS:Porcine autologous BM-MNCs were isolated by gradient density centrifugation,and MSCs were obtained by adherence method.Prior to transplantation,both BM-MNCs and MSCs were colloidal gold labeled.Except the infract model group,pigs in the other 3 groups were developed into AMI models by oppressing the left anterior descending branch with balloon catheter.Ninety minutes after modeling,(6.0±1.3)×107 autologous BM-MNCs and (4.5±2.1)x 107 MSCs were respectively transplanted into pigs in the BM-MNC group and the MSC group via the coronary artery and cultured for 28 days.MAIN OUTCOME MEASURES:Observation of pathological changes of cardiac muscle tissue by light and electron microscope;Examination of cardiac function by ultrasonograph;Detection of the number of blood vessels and apoptotic myocardial cells,and expression of nuclear factor-κB (NF-κB) and troponin Ⅰ and its correlation to cardiac function by immunohistochemistry;Detection of mRNA expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in the cardiac tissue as well as its correlation to cardiac function by reverse transcription-polymerase chain reaction (RT-PCR).RESULTS:In the MSC group,there was proliferation of a great deal of blood vessels as well as growth of abnormal cell masses around the coronary vessels,while the BM-MNC group exhibited the "budding" of many capillary vessels.Prior to transplantation,cardiac function indices were basically similar among each group (F = 1.550,P＞0.05).Twenty-eight days after transplantation,left ventricular ejection fraction was significantly lower in the control,BM-MNC,and MSC groups than in the infarct model group (F = 5.30,P＜0.05),while endocardial fractional shortening was significantly higher (F = 10.67,P＜0.01).Compared with the infarct model group,the number of blood vessels in the infarct zone and infarct border zone was increased in the BM-MNC group (F=29.56-34.87,P＜0.01) and had no apparent change in the MSC group.In the BM-MNC and MSC groups,apoptotic myocardial cells in the infarct zone and infarct border zone were significantly reduced (F=14.31-35.34,P＜0.01 ) and troponin I expression rate was significantly increased (F=19.05,P＜0.01 ),as compared with the infarct model group.In addition,NF-κB positive rate in the infarct border zone was significantly lower in the BM-MNC and MSC groups than in the infarct model group (F=19.05,P＜0.01).VEGF gene expression level in the infarct border zone was significandy higher in the BM-MNC group than in the infarct model group and MSC group (F = 49.41,P＜0.01).bFGF gene expression level in the infarct border zone was significantly higher in the MSC group than in the infarct model and BM-MNC groups (F=4.71,P＜0.01).LVEF was negatively correlated to myocardial cell apoptosis rate and NF-κB level (r=-0.441 1,P＜0.05;r=-0.579 6,P＜0.01 ).LVEF was positively correlated to number of blood vessels,VEGF and bFGF expression (r=0.775,P＜0.01;r=0.565 1,P＜0.05;r=0.573 5,P＜0.05).CONCLUSION:Transplantation of both autologous BM-MNC and MSC via coronary artery can improve the condition of left ventricular remodeling subsequent to myocardial infarction.The improvement of cardiac functions is related to the increase of blood vessels,VEGF and bFGF expression,the decrease of myocardial cell apoptosis and NF-κ B level in cardiac muscle tissues after stem cell transplantation.BM-MNC transplantation better promotes blood vessel proliferation and VEGF expression in the cardiac tissue but produces worse effects on bFGF gene expression than MSC transplantation.

To produce recombinant Maxadilan using gene engineering technology, the gene of recombinant Maxadilan which expressed in protocaryon were designed and synthesized according to the amino acid sequences of Maxadilan. The recombinant plasmid pKYB-MAX was constructed and transformed into host bacteria Escherichia coli strain ER2566. After the MAX-intein-CBD fusion protein was purified by chintin-affinity chromatography, the self-cleavage activity of the intein was induced by beta-mercaptoethanol and the recombinant Maxadilan was released from the chitin-bound intein tag. The molecular weight of peptides was determined by the laser flight mass spectrometry and the results was conformity with the theoretical value. The biological activity analysis showed that recombinant Maxadilan significantly enhanced the concentration of serum glucose.
Animals ; Base Sequence ; Escherichia coli ; genetics ; metabolism ; Insect Proteins ; biosynthesis ; genetics ; Inteins ; genetics ; Isopropyl Thiogalactoside ; pharmacology ; Molecular Sequence Data ; Recombinant Proteins ; analysis ; biosynthesis ; genetics

External Guide Sequences (EGSs) represents a novel nucleic acid based gene interference approach to modulate gene expression. They are oligonucleotides that consist of a sequence complementary to a target mRNA and recruit intracellular RNase P for specific degradation of the target RNA. DNA-based EGS1386 with a size of 12 nt was chemically synthesized to target the mRNA coding for the UL49 gene of human cytomegalovirus (HCMV). The DNA-based EGS1386 molecule efficiently directed human RNase P to cleave the target mRNA sequence in vitro. A reduction of more than 50% in the levels of UL49 expression was observed in human cells treated with the DNA-based EGS1386 targeted UL49 assayed by fluorescent quantization PCR and Western blotting. This results showed that the DNA-EGS1386 can effectively guide the RNase P cut the target mRNA. Therefore, DNA-EGS can develop into a new gene silencing technology and potential of the anti-viral reagents.
Base Sequence ; Cytomegalovirus ; drug effects ; genetics ; metabolism ; Cytomegalovirus Infections ; enzymology ; virology ; DNA, Viral ; genetics ; Directed Molecular Evolution ; methods ; Gene Expression Regulation, Viral ; Humans ; Nucleic Acid Conformation ; Oligodeoxyribonucleotides ; genetics ; pharmacology ; RNA, Guide ; chemistry ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Ribonuclease P ; genetics ; metabolism ; Viral Structural Proteins ; genetics ; metabolism

Objective·To compare the effects and complications of carbon dioxide laser laryngectomy and open partial laryngectomy in the treatment of laryngeal carcinoma.Methods·Clinical data of 101 patients with stage Ⅰ-Ⅲ laryngeal squamous cell carcinoma admitted to Renji Hospital,Shanghai Jiao Tong University School of Medicine from January 2013 to January 2018 were selected and divided into open surgery group and minimally invasive group according to different surgical methods.The open surgery group received open partial laryngectomy(57 cases),and the minimally invasive group received carbon dioxide laser laryngectomy(44 cases).Follow-up visits were made by telephone and outpatient visits,and the operation time,postoperative laryngeal mucosa edema time,length of stay,5-year survival rate,complication rate and recurrence rate of the two groups were compared in the different clinical stages(stage Ⅰ,stage Ⅱ,and stage Ⅲ)and tumor locations(glottic type and non-glottic type).Results·The operation time,postoperative laryngeal mucosa edema time and length of stay of the minimally invasive group were shorter than those of the open surgery group(P＜0.05).Comparison of the 5-year survival rates of patients with the same clinical stage and glottic type between the two groups showed no statistically significant difference(both P＞0.05);while comparison of the 5-year survival rates of patients with non-glottic type between the two groups(62.5％in the open surgery group vs 0 in the minimally invasive group)showed a statistically significant difference(P＜0.05).In the minimally invasive group,there were 3 cases of complications,including 1 case of fever and 2 cases of lung infection;in the open surgery group,there were 14 cases of complications,including 5 cases of fever,7 cases of lung infection and 2 cases of pharyngeal fistula.The complication rate of patients in the minimally invasive group(6.82％,3/44)was lower than that in the open surgery group(24.60％,14/57),and the difference was statistically significant(P＜0.05);the complication rate of patients with stage Ⅱ and glottic type in the minimally invasive group was significantly lower than that in the open surgery group(P＜0.05).Comparing the recurrence rate of patients in the two groups(10.53％in the open surgery group vs 2.27％in the minimally invasive group),the difference was not statistically significant(P＞0.05).Conclusion·Compared with open partial laryngectomy,carbon dioxide laser laryngectomy has faster postoperative recovery and lower complication rate in the treatment of early and middle laryngeal malignant tumors,so it is worth promoting.