In order to assess whether gene transfection could be mediated by ultrasound in association with P85 and find the appropriate parameters of ultrasound irradiation, the effects of ultrasound with or without P85 on gene transfection of HepG2 cells were examined. The HepG2 cells were irradiated by ultrasound at 1 MHz, 0.4-2.0 W/cm(2) and 50% duty cycle with plasmid encoding enhanced green fluorescent protein (EGFP) as a report gene. Forty-eight h later, the expression of EGFP was detected under the fluorescence microscopy. Transfection efficacy was quantitatively assessed by flow cytometry, and cell viability was evaluated by trypan blue exclusion. The results showed that the transfection efficacy was increased with the increases in ultrasound output power and the ideal transfection efficacy was achieved in HepG2 cells irradiated by ultrasound at 0.8 W/cm(2) for 30 s. The transfection efficacy in ulstrasound+P85 group was three times higher than in single ultrasound group [(17.63+/-1.07)% vs (5.57+/-0.56)%, P<0.05]. The cell viability was about 81% and 62% in ultrasound group and ultrasound+P85 group respectively. It was concluded that ultrasound in combination with P85 could mediate the gene transfection of HepG2 cells, ideal transfection efficacy was achieved by ultrasound irradiation at 0.8 W/cm(2) for 30 s, and P85 could somewhat increase the damage to cells caused by ultrasound.
Cell Survival/genetics ; Green Fluorescent Proteins/genetics ; Hep G2 Cells ; Poloxalene/*pharmacology ; Transfection ; Ultrasonics

ObjectiveTo investigate the social support and its related factors of elderly patients with chronic obstructive pulmonary disease (COPD).Methods60 outpatients with COPD over the age of 60 years were recruited in the study. Social support rating scale (SSRS) and demographic information were applied for investigation.ResultsScores of objective, subjective support, and availability of support were lower than the norm (P<0.05). Social support was significantly different among elderly patients with age, education levels and matital status (P<0.05). Patients aged over 70 years, low education level, divorced or widowed were vulnerable groups with low social support.ConclusionEfforts should be made to strengthen the social support system for elderly patients with COPD, especially for those aged over 70 years, low education, divorced or widowed.

Objective To investigate the influencing factors and prognosis of early and late recurrence after radical resection of primary hepatocellular carcinoma(HCC).Methods The clinical data of 117 patients who received radical resection of HCC at the Tongji Hospital of Huazhong University of Science and Technology from January 2003 to December 2006 were retrospectively analyzed.Tumor recurrence occurred within 2 years after operation was defined as early recurrence,and tumor recurrence occurred latter than 2 years after operation was defined as late recurrence.Relationship between postoperative tumor recurrence and level of alpha-fetoprotein (AFP),AFP/V,tumor diameter,tumor number,blood vessel invasion,tumor differentiation,hepatic cirrhosis,hepatic function,hepatitis B surface antigen,procedure of hepatic resection and blood transfusion was analyzed.The overall survival and disease-free survival rates were determined by Kaplan-Meier method,and the survival rate was analyzed by Log-rank test.Results Eighty-five(72.6％)patients were found with tumor recurrence,including 59(50.4％)with early tumor recurrence and 26(22.2％)with late tumor recurrence.Levels of AFP,AFP/V,tumor diameter,tumor number,blood vessel invasion,tumor differentiation and blood transfusion were the influencing factors of early recurrence(x2 = 12.78,13.40,5.79,9.98,10.26,9.48,8.32,P ＜ 0.05).Level of AFP and hepatic cirrhosis were the influencing factors of late recurrence(x2 =4.46,7.75,P ＜ 0.05).AFP/V,tumor number and blood vessel invasion were the independent risk factors of early recurrence(RR = 0.170,0.172,0.064,P ＜ 0.05).Hepatic cirrhosis was the independent risk factor of late recurrence(RR = 2.809, P ＜ 0.05).The 1-,3-,5-year overall survival rates and tumor-free survival rates were 82.6％,60.8％,54.9％ and 65.0％,38.5％,23.1％.There were significant differences in overall survival and disease-free survival rates among patients with AFP ＜20 μg/L,AFP/V ＜ 14 μg/(L · cm3)or AFP/V ≥ 14 μg/(L · cm3)(P ＜ 0.05).The 1-,3-,5-year overall survival rates of patients with early tumor recurrence were 64.9％,23.0％ and 20.5％,respectively,and the 1-,3-,5-year overall survival rates of patients with late tumor recurrence were 100.0％,88.5％ and 72.5％,respectively.A significant difference in the 1-,3-,5-year overall survival rates between patients with early or late tumor recurrence was observed(x2 = 26.918,P ＜0.05).Conclusions AFP/V,tumor number,blood vessel invasion were independent risk factors of early tumor recurrence,and hepatic cirrhosis was the independent risk factor of late tumor recurrence.There is a significant difference in the survival rate between patients with early or late tumor recurrence.

The value of color Doppler flow imaging (CDFI) and intravenous contrast-enhanced ultrasound (CEUS) for assessing the transplanted liver and early diagnosing complications by examining hemodynamic changes was discussed. Seventy-five patients with orthotopic liver transplantation (OLT) underwent CDFI. The following parameters were measured: peak systolic velocity (PS), resistance index (RI) and Doppler perfusion index (DPI) of the hepatic artery (HA), time average velocity (TAV) of portal vein (PV) and velocity of hepatic vein (HV) in different stages postoperation. And 11 patients of them received CEUS. Thirty healthy subjects were enrolled as controls. The results showed that: (1) In 23 patients without obvious complications, TAV of PV within 15 days post-operation was significantly higher than in controls (P<0.05), PS and DPI of HA within 7 days postoperation were lower, but RI was higher than in controls (P<0.05); (2) When the hepatic artery thrombosis (HAT) occurred, PS and DPI of HA were obviously decreased, but TAV of PV significantly increased like a high saw-tooth wave; (3) While rejection occurred, both TAV of PV and PS of HA were decreased with the increase in RI of HA, and the triphasic wave of HV disappeared and displayed as saw-tooth wave; (4) The incidence of biliary complications in liver transplantation was increased when DPI was reduced; (5) Seven cases of hepatic carcinoma relapse after OLT demonstrated hyperecho in the arterial phase and hypoecho in the portal and later phase on CEUS; (6) In 2 cases of HA thrombus, there was no visualized enhancement in arterial phase of CEUS, but enhancement during the portal vein and parenchymal phase. It was concluded that the hemodynamic changes of PV, HA and HV in the transplanted liver are valuable for assessing the transplanted liver and early diagnosing complications on CDFI and CEUS.
Bile Ducts/pathology ; Contrast Media/*administration & dosage ; Hemodynamics ; Hepatic Artery/pathology ; Infusions, Intravenous/*methods ; Liver Transplantation/*adverse effects ; Liver Transplantation/*methods ; Liver Transplantation/ultrasonography ; Perfusion ; Postoperative Complications ; Ultrasonography/*methods ; Ultrasonography, Doppler/*methods

Objective To observe the effects of electroacupuncture at Ganshu (BL18) and Zusanli (ST36) on depression state and gastrointestinal motility and relevant protein expressions in brain of rats with stress-induced depressive functional dyspepsia. Methods Totally 48 Wistar rats were randomly divided into normal, model, electroacupuncture (EA) and sham-operation group, with 12 rats in each group. Depressive functional dyspepsia model was established by chronic unpredictable mild stress protocols. From day 15 to day 28, the EA group was given electroacupuncture at BL18 and ST36, and the sham-operation group was stimulated by electroacupuncture at 1/3 and 2/3 of the tail. Body weight test, open field test and sucrose preference test were measured on day 0, day 14 and day 28. After 14-day administration, gastric residual volume and intestinal propulsion rate of semisolid paste were detected. The expression of βCaMK inⅡ lateral habenulas and BDNF in hippocampus were measured by Western blot. Results Compared with the normal group, the weight, the sucrose preference rate, the behavior score, the small intestinal propulsion rate of semisolid paste and the expression of BDNF in hippocampus decreased significantly both in the model group and sham-operation group (P<0.01), and the gastric residual volume of semisolid paste and the expression of βCaMK inⅡ lateral habenulas increased significantly (P<0.01). Compared with the model group, the weight, the sucrose preference rate, the behavior score, the small intestinal propulsion rate of semisolid paste and the expression of BDNF in hippocampus increased significantly in EA group (P<0.01), and the gastric residual volume of semisolid paste and the expression of βCaMK inⅡ lateral habenulas decreased significantly (P<0.01). Compared with the sham-operation group, the expression of βCaMKⅡ in lateral habenulas decreased significantly, the expression of BDNF in hippocampus increased significantly in EA group (P<0.01). Conclusion Electroacupuncture at Ganshu (BL18) and Zusanli (ST36) can improve depression and gastrointestinal motility of rats with depressive functional dyspepsia, which may be related to the inhibition of βCaMK inⅡ lateral habenulas and the activation of BDNF in hippocampus dyspepsia.

Purpose　To study myocardial pathology and its pathogenesis in diabetes. Methods　Myocardial structure of alloxan induced diabetic rats was observed under light microscopy (LM)and transmission electron microscopy(TEM).Activity of superoxide dismutase(SOD),glutathione peroxidase (GSH-Px)，nitric oxide synthase(NOS)and content of malondialdehyde (MDA),nitric oxide(NO) were detected biochemically in myocardial homogenate. Results　Atrophy and degeneration of myocardium and interstitial fibrosis were found under LM and expansion of mitochondria and smooth endoplasmic reticulum, destruction of myofibril and interstitial proliferation of collogen fiber under TEM. Activity of SOD and GSH-Px decreased significantly, while content of NO and MDA and activity of NOS increased significantly in diabetic rats. Conclusion　Atrophy of myocardium, expansion of mitochondria,destruction of myofibril and interstitial fibrosis are the main morphological changes of diabetic cardiomyopathy, and lipid peroxidation and NO may be involved in it.

0.05). Second and third generation of the implanted tumor were obtained in 8 nude mice respectively,and all transplanted tumors survived. Liver,bone and lung metastasis were found in 100% of nude mice between 10 w～14 w. [WT5”HZ] Conclusions This nude mouse HCC model holds the biologic features of human HCC and serves the purpose of in vivo study investigation .

This study examined the effect of P85 (a pluronic block copolymer) and microbubble (MB) ultrasound contrast agents under ultrasound irradiation on gene transfection and expression. The pEGFP plasmids that can encode enhanced green fluorescent protein (pEGFP) served as a report gene and were mixed with different concentrations of MB/0.05% (w/v) P85. Then the plasmids were transfected into human hepatoma G2 (HepG2) cells. The HepG2 cells treated with MB/P85 or without treatment were exposed to ultrasound (US parameters: 1 MHz, 1.0 W/cm(2), 20 s, 20% duty cycle). Twenty-four hours later, the transfection efficiency was assessed by fluorescence microscopy and fluorescence activated cell sorting (FACS) analysis. The cell viability was evaluated by Trypan blue exclusion test. The results showed that the gene transfection efficiency in HepG2 cells under ultrasound irradiation was significantly higher than that without ultrasound irradiation. HepG2 cells in the MB or P85 group in the absence of ultrasound expressed less amount of green fluorescent protein. The expression efficiency reached (22.14±3.06)% and the survival rate was as high as (55.73±3.32)% in the 30% MB plus P85 group. It was concluded that MB and P85 in the presence of ultrasound can enhance gene transfection and expression.

OBJECTIVE:To optimize the processing technology of rehmanniae radix praeparata. METHODS:Using transfer rates of catalpol,rehmaionoside D,acteoside,isoacteoside,polysaccharide as indexes for comprehensive score,heating tempera-ture(pressure),heating time and heating times as investigating factors,L9(34)orthogonal test was used to optimize the processing technology of rehmanniae radix praeparata,and verification test was conducted. RESULTS:The optimal processing technology of rehmanniae radix praeparata was as follow as heating temperature of 125 ℃,pressure of 150 kPa for twice,2 h every time. The comprehensive scores of 3 batches of samples were 0.6985,0.6755,0.7016 in the verification test,respectively,RSDs were less than 5%(n=3). CONCLUSIONS:Optimized processing technology is simple,stable,feasible,and can provide reference for in-dustrial production of rehmanniae radix praeparata.

AIM: To study the protective effects of the extract of Ginkgo biloba (EGB) on brain tissues in experimental diabetic rats and explore its possible mechanisms. METHODS: Thirty male Sprague-Dauley rats were divided into three groups: normal control group, diabetic group and EGB-treated group. Strepozotocin were injected intraperitoneally on the later two groups to induce diabetes, EGB-treated group was injected intraperitoneally with EGB, and the others were treated with normal saline at the same volume. After five weeks, the content of endothelin (ET), malondial dehyde (MDA), NO2~-/NO 3~-, and the activity of superoxide dismutase (SOD), nitric oxide synthase (NOS) were determined in brain homogenate, and the level of blood glucose, insulin and ET were measured respectively. In addition, the morphologic changes of the brain tissue were studied by light microscopy and electron microscopy, respectively. RESULTS: In diabetic group, there were degeneration of neuron, brain edema, softened focus and demyelination in the white matter of brain by light microcopy. There were expansion of mitochondria of neuron and gliocyte, the shortened of crista, demyelination of neurofiber and injury of blood-brain-barrier by the electron microscopy. After treated with EGB, the pathological changes decreased in brain under light microcopy and electron microcopy. Compared with diabetic rats, the activity of SOD and the level of serum insulin increased, while the level of blood ET, the activity of NOS, the content of ET, MDA, NO2~-/NO3~- decreased in EGB-treated rats (P