OBJECTIVETo construct microRNA-223 overexpression and suppression lentivirus vectors and determine their effects after infecting oral squamous cell carcinoma (OSCC) cell line.

METHODSLentivirus vectors GV229 and GV232 were cut by the restriction sites of Age I and EcoR I and connected to the target gene, which contained mature microRNA-223 and microRNA-223 oligonucleotide. Real-time polymerase chain reaction (PCR) method was used to detect the microRNA-223 expression level after infecting the recombinant lentivirus vector into the OSCC cell line.

RESULTSThe successful construction of microRNA-223 recombinant lentivirus vectors was confirmed by the PCR method and DNA sequencing. HN-30 cell infected with microRNA-223 overexpression vector showed a significant increased in microRNA-223 expression, whereas HN-30 cell infected with microRNA-223 inhibitor vector suppressed microRNA-223 expression.

CONCLUSIONThe microRNA-223 overexpression and suppression lentivirus vectors are successfully constructed. These vectors could alter the expression level of microRNA-223 in OSCC cell line significantly, and provide a stable cell line for functional studies in the future.

Base Sequence ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Lentivirus ; MicroRNAs ; metabolism ; Real-Time Polymerase Chain Reaction

Objectives To provide reference for the improve of health care system and healthcare insurance con-trolling method, through researching on the value-based healthcare paying system in South Korea. Methods The basic characteristics of the Korean health care system was researched, Specifically, its reform experiences of value-based health insurance review and assessment paying system,which government took. Results South Korea’s health care pay-ing system had achieved both good social and policy effects. Conclusions Value-based health care paying system is the main reason for the reducing of health care expenditures and making of scientific health policy.

Aged ; Humans ; Lower Extremity ; Middle Aged ; Muscle, Skeletal/physiology* ; Vibration

AIM: To investigate the behaviour of 3T3 fibroblast and macrophage co-culture on blood fibrin clot or adipose tissue with recombinant basic fibroblast growth factor (rhbFGF). METHODS: MTT method, inverted contrast microscopy, Giemsa staining as well as scanning electron microscope were used in the present study. RESULTS: The effect of rhbFGF on co-culture of 3T3 fibroblast and mouse macrophage on blood fibrin clot in low-serum DMEM with rhbFGF were monitored, and 3T3 fibroblast and macrophage growed well on the blood fibrin blot in low-serum DMEM with rhbFGF. CONCLUSION: The blood fibrin clot, with its low immunogenicity, could be used as a bionic support for three-dimensional tissue culture, and also a physiological carrier to distribute the growth factor rhbFGF for the cells.

Objective:To observe the effect of horse-riding squat exercise plus Governor Vessel-regulating Tuina(Chinese therapeutic massage)therapy on static balance function in patients with stroke.Methods:A total of 176 stroke patients were enrolled as the study subjects and divided into four groups by the random number table method,namely horse-riding squat+Tuina group,horse-riding squat group,Tuina group,and control group,with 44 cases in each group.The control group was treated with rehabilitation balance training.The horse-riding squat+Tuina group,horse-riding squat group,and Tuina group were treated with additional horse-riding squat exercise plus Tuina,horse-riding squat exercise,and Tuina treatment,respectively.Four weeks of treatment was regarded as one treatment course.After 1 treatment course,the balance function of the four groups was compared.Results:After treatment,the ellipse area of motion and the length of motion in all four groups were reduced,and the intra-group differences were all statistically significant(P＜0.05).The difference in the ellipse area of motion before and after treatment in the horse-riding squat+Tuina group and the horse-riding squat group was larger than that in the control group,and the differences were statistically significant(P＜0.05).The difference in the ellipse area of motion before and after treatment in the horse-riding squat+Tuina group was larger than that in the Tuina group,and the difference was statistically significant(P＜0.05).The difference in the length of motion before and after treatment in the horse-riding squat+Tuina group,the horse-riding squat group,and the Tuina group was greater than that in the control group,and the differences were statistically significant(P＜0.05).There was no statistically significant difference in the pairwise comparison between the horse-riding squat+Tuina group,the horse-riding squat group,and the Tuina group(P＞0.05).Conclusion:Horse-riding squat exercise plus Governor Vessel-regulating Tuina therapy can effectively improve the static balance function in patients with stroke.Training the affected lower limb weight-bearing exercise,adjusting the center of gravity distribution,and promoting the mutual balance and coordination between muscle groups may be the mechanism of improving the static balance function.

ObjectiveTo establish a new method for preparing synaptosomes.MethodsDensity gradient centrifugation method was used to isolate synaptosomes of mouse, checking by transmission electron microscopy.ResultsSynaptosomes prepared by this method had intact morphological characteristics, surrounding with a continuous oval-shaped membrane structure, moreover, mitochondrion and lots of synaptic vesicle in them.ConclusionThis method is applicable to establish a rapid, convenient and useful method for preparing synaptosomes.

Objective:To analyze the relationship between hepatitis B virus(HBV)gene mutation at 1896 in precore region with genotype and replication of HBV and the liver function of patients.Methods:HBV precore 1896 site mutation,the genotype of HBV and serum content of HBV DNA were determined by PCR in 60 patients positive of HBV DNA.Chemiluminescence miacropaticle immunoassay(CMIA)was used for detection of serum HBeAg and HBeAb.Liver function parameters were ob- tained by routine biochemistry method.Results:The alanine aminotransferase(ALT)level in HBV with 1896 site mutation was significantly higher than that in the wildtype virus.Site mutation at 1896 had no correlation with HBeAg,HBV genotype and HBV DNA content.HBV DNA content in patient with genotype C was significantly higher than that with genotype B(P

Objective To observe the effects of Dahuangfuzi decoction on the intestine barrier functional of acute necrotizing pancreatitis in rats. Methods The 60 rats were randomly divided into sham operation group ( n = 19 ), ANP group ( n = 21 ), and Dahuangfuzi treatment group ( n = 20). The rats of ANP group were induced by injecting 1 ml/kg of 4% sodium taurocholate into the pancreatiobiliary duct, and jejunal fistula was esablished. The rats of treatment group received Dahuangfuzi decoction (2 ml, repeated at 4 and 8 h)through jejunum distal stoma tube 0. 5 h after ANP induction. The other 2 groups received same amount of normal saline. Blood sample was collected through abdominal aorta, 24 h after ANP induction, and the serum amylase, endotoxin, D-lactate, plasma diamine oxidase (DAO) were detected. Pancreas, small intestine tissue was harvested for pathologic examination, index of intestinal epithelial damage was measured and ultrastructural changes in small intestinal mucosa was observed. Results The expression of serum amylase, endotoxin,D-lactate, DAO in sham operation group was ( 152 ± 32 ) U/L, (6.95 ± 2.10) pg/L, ( 3.96 ± 1.08 ) μg/mland ( 14.26 ± 2.67 ) μg/ml, while the corresponding values were ( 1549 ± 93 ) U/L, (40.48 ± 3.41 ) pg/L,( 12.34 ± 1.23 ) μg/ml and ( 80.28 ± 3.54) μg/ml in ANP group, and they were (655 ± 49 ) U/L, ( 19.55 ±2.50) pg/L, (6.75 ± 1.36 ) μg/mland ( 20.69 ± 7.53 ) μg/ml in treatment group. The values in ANP group were significantly higher than those in sham operation group. The values in treatment group were significantly lower than those in ANP group, but significantly higher than those in sham operation group ( P ＜ 0.05 or P ＜0. 01 ). The thickness and height of intestinal mucosa in ANP group were ( 389.44 ± 29.87 )μm and ( 16.52 ±3.73) μm, which were significantly lower than those in treatment group [(501.95 ± 45.38 )μm, (27.82 ±5.17)] μm, and in sham operation group [( 658.72 ± 57.49 ) μm, ( 35.49 ± 6.43 )μm, Index of intestional epitholial donage in ANP group was 3.72 ± 0.65 which is significently higher than those in theatment (2.12 ±0.37 ) and in sham operation group (0.85 ± 0.24). The intestinal mucosa histological and ultrastructural changes in Dahuangfuzi treatment group were better than those in ANP group. Conclusions Dahuangfuzidecoction can significantly decrease the damage of intestine barrier function in ANP rats.

OBJECTIVETo observe the three-dimensional distribution of vessels, and to establish a new method for measurement of blood flow velocity in mice cerebral cortex using two-photon laser scanning microscopy and fluorescence probe labeling technique.

METHODSThe mouse was made cranial window surgery and injected Texas-Red through tail vein after anesthetized. The three-dimensional imaging of vessel was obtained through z-stack scanning, and blood flow velocity was quantified through line scanning.

RESULTSWe could detect vascular distribution for more than 500 µm depth using two-photon microscopy. The velocity of blood flow was (0.59 ± 0.12) mm/s in capillary.

CONCLUSIONThe method for observing the brain blood flow by two-photon microscopy was established, which could achieve quantification of single vascular blood flow velocity and provide experimental evidence for basic research and medical applications.

Animals ; Blood Flow Velocity ; Brain ; blood supply ; Capillaries ; Cerebrovascular Circulation ; Fluorescent Dyes ; Hemodynamics ; Mice ; Microscopy, Fluorescence

Objective To investigate the effects of cholinergic pathway on acute renal tubular cell injury induced by acute oxygen and glucose deprivation. Methods Rat kidney macrophages were isolated and cultured for constructing macrophages and renal epithelial cells co-cultivating model of oxygen-glucose deprivation (OGD), and the model cells were divided into three groups: OGD alone group, acetylcholine (ACh 100μmol/L)+OGD group and ACh + galantamine (Gal 10μmol/L)+OGD group. The cells underwent OGD treatment for 1 hour, and normally cultured for 24 hours. The expressions of TNF alpha, IL-1 beta, and IL-10 in supernatant fluid were detected by ELISA, the renal tubular cell viability was determined by MTT assay, the expression of acetylcholine esterase (AChE) mRNA and protein were determined by RT-qPCR and Western blotting. The activity of AChE was determined by colorimetric method. Results The expressions of TNF alpha (pg/ml) in OGD, Ach+OGD group, Ach+Gal+OGD groups were 140.2±44.81, 119.46±4.42 and 103.31±1.62 respectively (P<0.05), those of IL-1β (pg/ml) were 172.26±13.51,144.34±5.53 and 119.37±11.42 respectively (P<0.05), and those of IL-10 (pg/ml) were 181.47±16.01, 173.62±10.12 and 188.36±8.73 respectively (P>0.05); The values of renal tubular cell proliferation were 55.02%±6.28%, 66.65%±6.47%, and 79.75%±4.22% respectively (P<0.01); the expressions of AChE mRNA in macrophages were 4.07±0.03, 4.22±0.15 and 3.98±0.29 respectively in the three groups (P>0.05); those of AchE protein were 0.66±0.07, 0.74±0.04 and 0.67±0.06 respectively (P>0.05); The activity of AChE (kU/L) was 0.51±0.02, 0.35±0.05 and 0.32±0.04 respectively (P=0.001, 0.001 and 0.368). Conclusions ACh and Gal could inhibit the secretion of inflammatory mediators and cholinesterase activity and can reduce the acute hypoxic renal tubular cell injury. The modulation of the cholinergic pathway in macrophages may be the important treatment method for acute renal injury in the future.