Objective To investigate the effects of Panax notoginseng saponins(PNS) on penile erection in rats with diabetes-as-sociated erectile dysfunction (ED) .Methods Ninety male Sprague-Dawley rats were established diabetic rats by injecting streptozo-tocin(STZ) ,and observing erectile phenomenon by injecting apomorphine .After 4 weeks of PNS treatment (low-dose ,medium-dose , high-dose group) ,erectile function in each group was assessed by intracavernous pressure (ICP) and mean arterial pressure(MAP) measurement .The level of nitric oxide(NO)and cyclic guanosine monophosphate(cGMP)in cavernous tissue were detected .Immu-nohistochemical staining and TUNEL were performed for detecting endothelial nitric oxide synthase (eNOS) and apoptosis ,respec-tively .Results ICP and ICP/MAP ratio were significantly increased in medium-dose and high-dose PNS treated groups compared with the diabetic untreated group(P<0 .05) .Compared with the diabetic untreated group ,the expression of eNOS and the levels of NO and cGMP were increased in medium-dose and high-dose PNS treated groups(P<0 .05) .The apoptosis ratecorpus of caverno-sum in 3 PNS treatment groups significantly decreased than the diabetic untreated group (P<0 .05) .Conclusion PNS can recovery the endothelial cell function in corpus cavernosum by adjusting the NO/cGMP pathway and controlling the accumulation of AGEs , and may be used for improving in diabetic ED rats .

Objective To study the effects of cripto on migration, invasion, and liver metastasis of colorectal cancer cell. Methods After human colorectal cancer cell line SW480 was transfected by cripto small interfering RNA (siRNA), the mRNA and protein level were determined by Real-time RT-PCR and Western blot, respectively. The migration and invasion ability were evaluated by wound-healing assay and boyden chamber model, respectively. Thirty nude mice model of liver metastasis from colorectal cancer was established by splenectomy. Results The siRNA could down-regulate the level of mRNA and protein of cripto in a dose- and time-dependent manner. Suppression of cripto expression could inhibit migration and invasion ability of human colorectal cancer cell in vitro. The metastastic rate and tumor nodules were lower in transfection with cripto siRNA than in two control groups in vivo. Conclusions Cripto gene might play an important role in regulation of liver metastasis from colorectal carcinoma cell, and suppression of cripto gene by siRNA can inhibit liver metastasis of colorectal cancer.

The stability of the intraocular lens ( IOL ) after cataract surgery is composed of decentration, tilt, rotation, and the change of anterior chamber depth. Its stability is an important factor affecting postoperative visual quality. By analyzing the related factors which influence the stability of intraocular lens, improvements can be identified for future cataract operations. The stability of intraocular lens is influenced by many factors: intraocular structure, the size and the symmetry of intraoperative capsulorhexis, the position of the intraocular lens, the material and design of the intraocular lens, etc. In order to improve the patient's vision, cataract surgeries have been experiencing an evolution. IOL material have also been contributing to such innovations.

Objective To investigate the expression and clinical significance of cell division cycle 6 (CDC6)and homeobox gene A5(HOXA5)in esophageal squamous cell carcinoma. Methods The expres-sion of CDC6 and HOXA5 in 51 specimens esophageal squamous cell carcinoma and 27 normal specimens esophageal tissues were evaluated by immunohistochemistry. Analyzed the relationship among the expression of CDC6 and HOXA5 protein and the clinicopathologic features of esophageal squamous cell carcinoma,along with the correlation between these two proteins. Results Immunohistochemical results showed that the positive expression rate of CDC6 and HOXA5 in esophageal squamous cell carcinoma tissue were 66. 7%(34 / 51)and 60. 8%(31 / 51),respectively,significantly higher than those in normal esophageal tissue18. 5%(5 / 27), 22. 2%(6 / 27),(χ2 = 16. 370,P = 0. 000;χ2 = 10. 528,P = 0. 001);There were significant positive correlation in the expression of CDC6 and HOXA5 and histological type(χ2 = 9. 031,P = 0. 011;χ2 = 7. 372,P = 0. 000), TNM stage(χ2 = 10. 474,P = 0. 015;χ2 = 11. 667,P = 0. 009),and there were no correlation in the expression of CDC6 and HOXA5 and age(χ2 = 0. 000,P = 1. 000;χ2 = 0. 001,P = 0. 972),sex(χ2 = 0. 049,P = 0. 824;χ2 = 0. 107,P = 0. 743),lymph node metastasis(χ2 = 3. 186,P = 0. 074;χ2 = 2. 212,P = 0. 137)in esophageal squamous cell carcinoma tissues. The expression of CDC6 and HOXA5 showed a positive correlation( r =0. 454,P = 0. 001). Conclusion The positive expression rate of CDC6 and HOXA5 in esophageal squamous cell carcinoma tissue were significantly higher than in normal esophageal tissue and close correlation with TNM stage and differentiation. High expression of CDC6 and HOXA5 may play important roles in the occurrence, development and proliferation of esophageal squamous cell carcinoma.

Objective: To adjust the distance between SD sequence and ATG in the same expressive plasmid pLX1 to enhance expression of heterologous bFGF gene in E. coli. Methods: Adjusting the different distance between SD sequence and ATG by Klenow and Mung Bean Nuclease. SDS PAGE and Western blot showed the expressed protein bFGF in E.coli. bFGF proteins were purified by HPHIC, HPGFC and HAC. Biological activity was examined by MTT. Results: Recombinant plasmids pLX2, pLX3 were obtained and the expressive levels were 8.03%, 9.9% respectively. Also the purified bFGF was obtained by HPHIC, HPGFC, HAC and its ED 50 was 2.29 ng/ml. Conclusion: Increasing the bFGF gene dosage by adjusting the distance between SD sequence and ATG could increase the expression level of a desired protein.

The main objective of this research is to investigate the effects of astragaloside IV, calycosin separately glucoside, formononetin on oxidative stress in Chang Liver cells induced by H2O2. In the experiments, Chang Liver cells (a kind of normal human hepatocytes) were used as the research object, bifendate which has a clear hepatoprotective effect was used as the positive control drug, then the oxidative damage model of Chang Liver cells were established by H2O2. Cells were divided into six groups: blank control group, oxidative stress group, astragaloside IV group, calycosin separately glucoside group, formononetin group and positive control group. Then endogenous antioxidant system related indexes were detected by micro plate and colorimetric method; intracellular reactive oxygen species (ROS) were detected by DCFH-DA fluorescent probe; and the expressions of CYP2E1 were evaluated by liver microsomes, mRNA, and protein, respectively with spectrophotometry, Real-time PCR method, and Western blot technique. Results showed that H2O2 decreased antioxidant activity, and increased ROS level and expression of CYP2E1. The above oxidative stress status had been changed with protections of the three components of Astragalus membranaceus (compared with oxidative stress group, P < 0.05, P < 0.01), which taken as a whole had equivalent effects as the drug of positive control group( bifendate). Taken together, three Astragalus membranaceus ingredients all had significant or extremely significant inhibiting effects on oxidative damaged Chang Liver cells which were induced by H2O2, and the oxidative damage of Chang Liver cells had been relieved.
Astragalus membranaceus ; chemistry ; Cells, Cultured ; Cytochrome P-450 CYP2E1 ; metabolism ; Humans ; Isoflavones ; pharmacology ; Liver ; drug effects ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Saponins ; pharmacology ; Triterpenes ; pharmacology

OBJECTIVETo study the protective effect of astragaloside IV on oxidative damages of Chang Liver cells induced by ethanol and H2O2.

METHODThe alcoholic and nonalcoholic oxidative damage models were established on Chang Liver cells with ethanol and H2O2, respectively. The cells viabilities were detected by MTT assay, transaminase activity and antioxidant ability were detected by micro plate and colorimetric method, reactive oxide species (ROS) was detected by DCFH-DA fluorescent probe and cell cycle was detected by flow cytometry. DNA ladder method was used to detect apoptosis.

RESULTBoth kinds of oxidative damage could decrease the viability and antioxidant enzyme activity of Chang Liver cells, and increase the transaminase activity and MDA content of extracellular fluid. The protective effects of astragaloside IV against those two kinds of oxidative damages were significant or extremely significant. Meanwhile, ethanol could decline the level of ROS significantly in the damaged cells, while H2O2 could increase it significantly. And the effect of astragaloside IV was to make ROS return to the normal level. Retardation of cell cycle progression of Chang Liver cells in G0/G1 induced by ethanol or H2O2 was relieved, and apoptosis was also inhibited.

CONCLUSIONAstragaloside IV had protective effect on oxidative damages of Chang Liver cells induced by ethanol and H2O2.

Antioxidants ; metabolism ; Apoptosis ; Cell Cycle ; drug effects ; Cell Line ; Cell Survival ; drug effects ; Ethanol ; pharmacology ; Humans ; Hydrogen Peroxide ; pharmacology ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Saponins ; pharmacology ; Triterpenes ; pharmacology

Aim To establish a model of the vascular endothelial cell (VEC) injury in SDrats.Methods SD rats were randomly divided into the control and the modelgroups. The model rats were injected with adrenaline diluted to 2. 5 times 0. 05 mg?100 g-1 (tid) for 5 d continously. From the 4th d, they were irritated for 5 min in the0℃ cold-water in the middle between adrenaline injections.The control rats weregiven 0. 9% NS as above. At 6th d, blood samples were taken from carotid arteries ofthe rats and the CEC counts, t - PA、PAI activities, 6-keto-PGF1? concentrations andthe platelet aggregation rate(max) were detected respectively. Results In the modelgroup, as compared with those in the control group, t - PA activity and 6-keto-PGF1?concentration decreased significantly(P

Objective To study morphological changes of rabbit artery endothelial cell injury and atherosclerosis caused by high fluoride and the role of selenium. Methods Twenty healthy male New Zealand white rabbits, body weight (2.0 ± 0.5)kg, were randomly divided into control group(drinking deionized water, fed basic diet), fluoride group(drinking fluoride 100 mg/L deionized water, fed basic diet), selenium group(drinking selenium 1 mg/L deionized water, fed basic diet), fluoride plus selenium group(drinking fluoride 100 mg/L deionized water, selenium 1 mg/L of deionized water, fed basic diet). The experimental period was 6 months. At 0, 3, 6 months of the experiment, serum fluorine and selenium levels were determined. At the end of the experiment,thoracic aorta was collected to observe its pathology and ultrastructural changes. Results Serum fluoride was significantly higher at the 3rd and the 6th month of experiment(all P ＜ 0.01 ) in fluoride group[ (0.589 ± 0.146),(0.772 ± 0.175)mg/L] and fluoride plus selenium group[ (0.502 ± 0.094), (0.693 ± 0.158)mg/L] than in control group[ (0.174 ± 0.002), (0.208 ± 0.031 )mg/L] and serum fluoride was significantly higher at 6 months than at 3 months(P ＜ 0.05 ) in fluoride group. Serum selenium was significantly higher at the 3rd and the 6th month of experiment (all P ＜ 0.01 ) in selenium group[ (0.252 ± 0.022), (0.319 ± 0.052)mg/L] and fluoride plus selenium group[ (0.239 ±0.016), (0.294 ± 0.018)mg/L] than in control group[(0.135 ± 0.014), (0.167 ± 0.019)mg/L], and serum selenium was significantly higher at the 6th month than at 3rd month of experiment in selenium group(P ＜ 0.05). Endothelial cell apoptosis indices were (4.92 ± 1.32)%, (30.30 ± 6.80)%, (6.57 ± 2.14)% and (14.29 ± 2.99)%, respectively in control group, fluoride group, selenium group and fluoride plus selenium group. Their main effect of fluorine and selenium was statistically significant (F = 106.833,20.082, all P ＜ 0.01 ). There were antagonistic effect between fluoride and selenium(F = 30.402, P ＜ 0.01 ). Pathological changes of rabbit aortic endothelial cells in fluoride group included endothelial with attached fibrin and red blood cells, and structural of the cells changed, with serious vascular injury; in fluoride plus selenium group apoptosis of endothelial cells decreased, with reduced number of attached red blood cells and fibrin, endothelial cell structure normal, the extent and scope of vascular damage significantly reduced. Conclusions Appropriate amount of selenium inhibits the apoptosis of endothelial cells induced by high fluoride, reduces aortic structural damage caused by high fluoride, and maintains the integrity of endothelial cells, thereby antagonizes the vascular damage and atherosclerosis induced by high fluoride.

Objective To compare the clinical efficacies of totally laparoscopic and laparoscopy-assisted radical total gastrectomies.Methods The retrospective cohort study was conducted.The clinicopathological data of 373 patients with gastric cancer who underwent totally laparoscopic or laparoscopy-assisted radical total gastrectomies from the 7 medical centers in China (82 patients in the Affiliated Hospital of Qinghai University,80 in the Traditional Chinese Medicine Hospital of Guangdong Province,60 in the First Affiliated Hospital of Xiamen University,51 in the Hangzhou First People's Hospital,46 in the First Affiliated Hospital of Xi'an Jiaotong University,30 in the Second Affiliated Hospital of Jilin University and 24 in the Xijing Hospital of the Fouth Military Medical University) between January 2015 and December 2016 were collected.Of 373 patients,the 183 and 190 patients were respectively divided into the totally laparoscopic group (undergoing totally laparoscopic radical total gastrectomy) and laparoscopy-assisted group (undergoing laparoscopy-assisted radical total gastrectomy),including 63 and 19 in the Affiliated Hospital of Qinghai University,36 and 44 in the Traditional Chinese Medicine Hospital of Guangdong Province,25 and 35 in the First Affiliated Hospital of Xiamen University,20 and 31 in the Hangzhou First People's Hospital,10 and 36 in the First Affiliated Hospital of Xi'an Jiaotong University,17 and 13 in the Second Affiliated Hospital of Jilin University,12 and 12 in the Xijing Hospital of the Fouth Military Medical University.Routine five-port method was applied for laparoscopic radical total gastrectomy and D2 lymphadenectomy.Roux-en-Y anastomosis was applied for digestive tract reconstruction,and digestive tract reconstruction was performed under laparoscopy in the totally laparoscopic group and via upper abdominal median incision in the laparoscopy-assisted group.Observation indicators:(1) operation and postoperative situations;(2) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect the postoperative overall survival and tumor recurrence or metastasis up to March 2017.Measurement data with normal distribution were represented as x±s.Comparison between the groups was analyzed by the t test.Measurement data with skewed distribution were represented as M (range).Comparisons of count data were analyzed using the chi-square test and Fisher exact probability.Results (1)Operation and postoperative situations:all the patients in the 2 groups underwent successful operations,without perioperative death.Esophagojejunostomy methods of 183 patients in totally laparoscopic group:conventional circular stapler method were performed in 28 patients,anti-puncture circular staplar method in 6 patients,OrVilTM method in 5 patients,functional end-to-end esophagojejunostomy method in 65 patients and peristalsis side-to-side esophagojejunostomy method in 79 patients.Conventional circular stapler method was applied to 190 patients in the laparoscopy-assisted group.Operation time,time of esophagojejunostomy,length of assisted incision,using time of analgesics and expenses of digestive tract reconstruction were (238± 55)minutes,(29±9)minutes,(5.1 ± 1.1)cm,(2.2±l.0)days,(18 332±2 141)yuan in the totally laparoscopic group and (217±39)minutes,(26±7)minutes,(7.8 ±2.0)cm,(2.7± 0.9)days,(16 237 ± 1 923)yuan in the laparoscopy-assisted group,respectively,with statistically significant differences between the 2 groups (t =4.324,3.455,-16.835,-5.561,9.949,P＜0.05).The cases with postoperative overall complications,anastomosis leakage,anastomosis stricture,anastomosis bleeding and expenses of esophagojejunostomy were respectively 24,9,7,5,(9 668±2 814)yuan in the totally laparoscopic group and 24,8,9,6,(9 331 ±2 067)yuan in the laparoscopy-assisted group,with no statistically significant difference between the 2 groups (x2 =0.036,0.107,0.189,0.059,t=1.322,P＞0.05).All the patients with postoperative complications were cured by symptomatic treatment.(2) Follow-up and survival situations:of 373 patients,336 were followed up for 4-26 months,with a median time of 13 months,including 166 in the totally laparoscopic group and 170 in the laparoscopy-assisted group.During the follow-up,cases with overall survival,tumor recurrence and tunor metastasis were respectively 150,10,16 in the totally laparoscopic group and 154,9,16 in the laparoscopy-assisted group (10 and 9 patients in the totally laparoscopic and laparoscopy-assisted groups with simutaneous tumor recurrence and metastasis),showing no statistically significant difference between the 2 groups (x2 =0.075,0.010,P＞0.05).Conclusions Total laparoscopic and laparoscopyassisted radical total gastrectomies are safe and feasible,with equivalent overall outcomes and effects of esophagojejunostomy.Compared with laparoscopy-assisted radical total gastrectomy,the postoperative pain time of patients in total laparoscopic radical total gastrectomy is less,but there are longer time of esophagojejunostomy and higher expenses of digestive tract reconstruction.