Objective To explore the feasibility,efficiency and effect of wide-type p53 gene transfected into cervical cancer HeLa cells by ultrasound microbubble intensifier. Methods Based on the optimum parameter of ultrasound wave irradiation (0.5 W/cm2 ,30 s) selected in pre-experiment,HeLa cells were divided into 5 groups:(A)simple plasmid group,(B)plasmid-ultrasound group,(C)plasmid-ultrasound-microbubble group,(D)plasmid-liposome group,(E)blank control group. At 24-48h after transfection,the transfection efficiency was detected by fluorescence microscope,the mRNA expression of p53 was detected by RT-PCR,the cell cycle was detected by flow cytometry,and the inhibition rate of cells growth was detected by MTT assay. Results Fluorescence microscopy showed that the expression of green fluorescent protein in group C was higher than that in group Band group D (P

Objective To investigate the effects of cold preservation and reperfusion injury on the regen-eration of donor liver and to study the mechanisms. Methods Male SD rats were divided in to sham group (6 rats), UW 1 h group (48 rats) and UW 12 h group (48 rats). Liver tissue specimens were collected at different time points after orthotopic liver transplantation or sham operation. The morphology of liver tissue was observed via light microscope and transmission electron microscope. Proliferation of hepatocytes and sinusoidal endothelial cells (SECs) were assessed by a double immunostaining technique using antibodies against rat endothelial cell antigen-1 and bromodeoxyuridine (BrdU). Expression of vascular endothelial growth factor (VEGF) and its receptors, fins-like tyrosine kinase-1 (flt-1) and fetal liver kinase-1 (flk-1) was evaluated by immunohistochemistry. The mRNA expression of flt-1 was detected by a RT-PCR method. Mean comparison in groups was conducted by one-way ANOVA or t test. Results BrdU labeling indexes of hepatocytes and SECs in UW 12 h group was significantly higher than those in UW 1 h group (F = 61.45,41.4, P < 0.05). The proliferation of hepatocytes peaked at 48 h after operation in both UW 1 h group and UW 12 h group. However, the proliferation of SECs was fallen behind compared to hepatocytes, with a peak appeared at 72 h in UW 1 h group and at 96 h in UW 12 h group, respec-tively. The expression of VEGF was up-regulated in both UW 1 h group and UW 12 h group compared to sham group. Furthermore, expression of flt-1 and flk-1 was found to be mainly limited in SECs, with a peak in expres-sion occurring between 72 h and 96 h, coinciding with the peak in SECs proliferation in UW 1 h group. Conversely, flt-1 was found to be reduced significantly on mRNA level at any time throughout the experiment in UW 12 h group compared to sham group (F = 141.67, P < 0.05). Conclusion Reduced expression of flt-1 results in a retarded regeneration of SECs, and then the recovery of rat donor liver function is delayed after cold-preserved transplantation.

This study was aimed to collect relevant provisions of bulging disease medical records in the Ming and Qing dynasties to excavate the syndrome differentiation, drug law and medication frequency analysis. A database was established to analyze relevant provisions. The results showed that the syndromes of liver stagnation and spleen deficiency, dampness-heat accumulation, spleen-yang deficiency are the most common type. The mainly used drugs are from the category to eliminate dampness and water, and to tonify the deficiency. The top three used drugs are poria, atractylodes, and dried citrus peel. It was concluded that medications used in the Ming and Qing dynasties were mainly targeted to the disordered zangfu-organ, which was mainly about the spleen and the stomach. The syn-drome differentiation is the mixture of deficiency and excess. And deficiency is the main part. The medication should combine tonification and reducing. And tonification should be paid attention to. The syndrome differentiation and treatment should be coordinated and the primary and secondary aspect should be identified. These rules provide ref-erence effect for the clinical practice and scientific research of bulging disease treatment.

Objective:To optimize the determination technology for the total amino acids in Runing pills. Methods:The process was optimized by L9 (34 ) orthogonal design using the hydrolysis temperature, hydrolysis time and hydrochloric acid concentration as the evaluation factors,and the content and yield of total amino acids as the evaluation indices. Results: The optimal determination condi-tions were as follows:the hydrolysis temperature was 110℃, the hydrolysis time was 22 h and the hydrochloric acid concentration was 6 mol·L-1 . Conclusion:The optimal determination technology can be applied in the total amino acids from Runing pills,which pro-vides the basis for the further research of Runing pills.

68 NIH Mice (24-25g) were divided into three groups. Group A was normal control, group B was treated with 54% alcohol in the beverages 0.25ml/per day, and group C was administrated 54% alcohol mixed with folic acid 0.25ml/per day. The histochemical reaction and ultrastructural changes in liver were observed. The reaction of glycogen (PAS reaction), Mg-ATPase SDH LDH G-6-Pasc and ACP was demonstrated respectively by Chayen's, Wachstein-Meisel, Lojda, Bancroft's method, and some procedures of these techniques were modified. The results showed that under the protection effect of acid folic, the regularities seemed to be present in the changes of the following histochemical reactions in the liver cells of the group C (treaded by acid folic) the PAS reaction and the activities of Mg-ATPase SDH LDH G-6-Pase were similar to the group A and apparently higher than that in group B. The activities of ACP were lower those in group B(P

Objective To observe the effect of intra-articular injection of dehydroepiandrosterone (DHEA)on the experimental osteoarthritis in rabbits and study the mechanism.Methods Forty rabbits un derwent unilateral anterior cruciate ligament transection(ACLT)and then divided into two groups randomly. 100?mol/L DHEA resolved in the dimethylsulphoxide were injected into the knees of experimental rabbits 4 weeks after transection,once a week for five weeks.Rabbits in the control group were treated under the same schedule using dimethylsulphoxide.All rabbits were killed 9 weeks after ACLT and the knee joints were evalu- ated by gross morphology and histology.The mRNA expression of metalloproteinases-3(MMP-3),tissue in- hibitor of metalloproteinases-1(TIMP-1)and interleukin-lbeta(IL-1?)in the cartilage and synovium was analyzed using reverse transcription polymerase chain reaction(RT-PCR).Results Gross morphologic in- spection and histological evaluation showed that the extent and grade of cartilage and synovium damage in the experimental group were less severe than the control group.The mRNA expression of MMP-3 in cartilage and synovium decreased significantly in the experimental group(both P＜0.05).The mRNA expression of TIMP-1 in cartilage and synovium increased significantly in the experimental group compared with that in the control group(both P＜0.05).No significant difference of IL-1?mRNA expression in cartilage was found between the experimental and the control groups(P＞0.05).The mRNA expression of IL-1?in the synovium was signifi- cantly suppressed in the experimental group compared with that in the control group(P＜0.01).Conclusion DHEA protects against cartilage degradation,alleviates synovium inflammation and inhibits the progression of osteoarthritis in the experimental model.Down-regulation of MMP-3 and up-regulation of TIMP-1 in cartilage and synovium and IL-1?in the synovium may be the mechanism of the protective effect of DHEA on os- teoarthritis.

Objective To evaluate the significance of cystatin C (CysC) in the diagnosis for early kidney injury of diabetes and hypertension by testing serum CysC and serum creatinine concentration and urinary albumin excretion (UAE) in patients with type 2 diabetes and primary hypertension.Methods Sixty-six patients with type 2 diabetes were selected,among whom the total 38 patients whose UAE were 30-300 mg/24 h were classified into early diabetic nephropathy (DN) group,and 28 patients whose UAE were ＜ 30 mg/24 h were classified into non diabetic nephropathy (NDN) group.Fifty-two patients with primary hypertension were selected,among whom the total 25 patients whose UAE were ≥ 30 mg/24 h were classified into hypertension with kidney injury group,and 27 patients whose UAE were ＜ 30 mg/24 h were classified into hypertension without kidney injury group.Sixty health people were selected as control group.The UAE,CysC and creatinine levels were detected.Results The CysC in early DN group was significantly higher than that in control group and NDN group [(1.84 ± 0.83)mg/L vs.(0.41 ± 0.62) and (0.66 ± 0.59) mg/L],and there was statistical difference (P ＜ 0.05).There was no statistical difference in CysC between NDN group and control group (P ＞ 0.05).There were no statistical differences in creatinine and UAE among the DN group,NDN group and control group (P ＞ 0.05).The CysC in hypertension with kidney injury group was significantly higher than that in control group and hypertension without kidney injury group [(0.93 ± 1.04) mg/L vs.(0.41 ± 0.62) and (0.69 ± 0.57) mg/L],and there was statistical difference (P ＜ 0.05).There was no statistical difference in CysC between hypertension without kidney injury group and control group (P ＞ 0.05).There were statistical differences in creatinine and UAE among hypertension with kidney injury group,hypertension without kidney injury group and control group (P ＞ 0.05).Conclusion CysC can be regarded as the value to judge early kidney injury,and it is very important inunderstanding patient's condition,by which appropriate treatment can be carried out to prevent or decrease the occurrence of chronic renal failure.

UNLABELLEDOBJECTIVE To study the in vitro effect and mechanism of Ginkgo biloba Extract 50 (GBE50) for inhibiting beta-amyloid (Abeta)-induced oxidative stress in rats' hippocampal neurons.

METHODSThe primary hippocampal neurons were cultured in vitro and divided into 4 groups, i. e. the normal control group (Ctrl), the Abeta group, the propanediol control group (PDO), and the six GBE50 concentrations groups (5, 10, 25, 50, 100, and 200 microg/mL). Excepted the Ctrl group, neurons were induced to oxidative stress by 20 gmolLAbeta25-35. The MTT and fluorescent probes labeling were used to observe the effect of GBE50 with different concentrations on the cell viability and the generation of intracellular reactive oxygen species (ROS) in neurons. Furthermore, Western blot was used to detect the cytoplasmic/total cytochrome C (Cyto C) ratio and total intracytoplasmal Cyto C, and the effect of the expression of oxidative stress-related protein Cyto C and activated Caspase-3 in three GBE50 concentrations groups (25, 50, and 100 microg/mL).

RESULTSCompared with the Ctrl group, the cell vitality was obviously lowered and intracellular ROS generation significantly increased after induction of 20 micromol/L Abeta25-35 (both P < 0.05). Compared with the Abeta group, the cell vitality was evidently improved after treated with different GBE50 doses. Except for 10 microg/mL, the cell vitality could be obviously elevated along with increased drug concentrations (P < 0.05). Meanwhile, the intracellular ROS generation decreased significantly in each GBE50 dose groups (P < 0.05). Abeta could increase the cytoplasmic/total Cyto C ratio and enhance the activated Caspase-3 expression significantly (P < 0.05). Compared with the Abeta group, among the three concentrations of GBE50, the Cyto C ratio was obviously lowered in the 100 microg/mL GBE50 group (P < 0.05), and the expression of activated Caspase-3 significantly decreased in 50 microg/mL and 100 microg/mL GBE50 groups (P < 0.05).

CONCLUSIONS20 micromol/L Abeta25-35 could induce the generation of intracellular ROS in hippocampal neurons. GBE50 could inhibit Abeta induced intracellular oxidative stress of neurons through lowering the cytoplasmic/total Cyto C ratio and inhibiting the activation of apoptosis protein Caspase-3 expression.

Amyloid beta-Peptides ; toxicity ; Animals ; Cells, Cultured ; Cytochromes c ; metabolism ; Hippocampus ; metabolism ; Neurons ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Peptide Fragments ; toxicity ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the feasibility of an intelligent three-dimensional ultrasound technique (Smart mid-sagittal planes,Smart MSP) in automatically detecting the fetal cranial mid-sagittal view.Methods Two hundred and forty pregnant women with singleton pregnancies were imaged to display the mid-sagittal view of fetal head using the Smart MSP by six doctors divided into three groups according the different experiences.Another two doctors were then invited to score the images acquired and to compare the successful rates of 6 doctors for acquiring fetal cranial mild-sagittal view according to the doctors' experience.Results The overall successful rate of acquiring the mid-sagittal view was 97.08％ (233/240) with the bipartial diameter plane (BPD plane) as the starting plane and 98.33(237/240) with the trans-cerebellum plane as the starting plane by using Smart MSP program.The successful rates in three groups were similar but the consumed times were different(P ＜0.05).The experienced doctors consumed less time than non-experienced doctors.The scores were also different in three groups.The scores of experienced doctors (group B and C) were significantly higher than that of non-experienced doctors (Group A) (P ＜0.05).There was no significant difference in the scores between experienced doctors (group B and C) (P＞0.05).Conclusions Smart MSP is a novel and feasible method for the automatic visualization of fetal cranial mid-sagittal plane,which may become an effective tool to screen the fetal midline anomalies in future.

The protective effect of vitamin E and the change in activity of enzymes at hepatic injury by carbon tetrachloride(CCl_4) were studied with histochcmical methods. 60 rats were devided into three groups(A,B,C). The group A was untreated control; the animals of group B and C were injected(i. m.) with 0.9% NaCl 1 ml/100g or vitamin E 1 ml/100g body weight qd for 7 days respectively and then treated with 30% CCl_4 0.05ml/l00g by adding to the solution of NaCl or vitamin E from 8th to 14th days. On the 15th day, rats were sacrificed for histochemical study of the liver parenchyma. The results showed that the activity of SDH, G-6-Pase, ATPase, 5-NT or AcP in liver lobules of groups B and C animals were changed conspicuously after treatment with CCl_4. The quantitative histochemistry of SDH and G-6-Pase was also consistent with these results. The activity of SDH, G-6-Pase, ATPase and 5-NT were inhibited, but of AcP was elevated. As compared with those in the rats of group A, the changes however in group B appeared more obviously than those in group C. The results indicated that CCl_4 of the given dose induced acute damage in liver cells, while vitamin E had important protective effect against acute liver damag induced by CCl_4.