Objective To explore the efficacy of pushen capsule on blood lipid and atherosclerosis in patients with ischemic stroke.Methods One hundred and twenty patients with ischemic stroke were randomly divided into three groups,treatment group (pushen capsule and rosuvastatin calcium),control group-1 (pushen capsule) and control group-2 (rosuvastatin calcium).There were 40 patients for each group.The course of treatment was 6 months.Data of serum total cholesterol (TC),triacylglycerol (TG),high density lipoprotein cholesterol (HDL-C),low density lipoprotein cholesterol (LDL-C),homocysteine (Hcy),high sensitive C reactive protein (hs-CRP),nitric oxide (NO),endothelin-1 (ET-1),the number and size of carotid plaques,and changes of intima-media thickness (IMT) were detected before and after treatment in three groups.The incidences of adverse cerebrovascular events and adverse events were observed in the three groups.And the therapeutic effects for dyslipidemia were evaluated in three groups.Results The treatment indicators were obviously improved after treatment in three groups (P ＜ 0.05).There were no significant differences in treatment indicators before treatment between three groups.The levels of TG and NO increased,the level of LDL-C,the size and thickness of carotid plaques decreased after treatment in control group-2 than those of control group-1.Levels of TC,LDL-C,Hcy,hs-CRP,ET-1,IMT,and the number,size and thickness of carotid plaques were significantly lower in treatment group than those of control group-1 (P ＜ 0.05),and the level of NO was higher in treatment group than that of control group-1 (P ＜ 0.05).Compared with control group-2,levels of TG,LDL-C,hs-CRP and the number,size and thickness of carotid plaques were significantly lower,and levels of HDL-C and NO were significantly increased in treatment group (P ＜ 0.05).There were no significant differences in the incidences of adverse drug reactions and adverse cerebrovascular events between the three groups (P ＞ 0.05).The total effective rate was higher in treatment group than that of control group-1 (P ＜ 0.05).There were no significant differences in total effective rates between treatment group and control group-2,and between control group-2 and control group-1 (P ＞ 0.05).Conclusion Pushen capsule combined with rosuvastatin calcium can obviously improve blood lipid metabolism in patients with ischemic stroke,which shows obvious antiatherosclerosis effect and good safety.

OBJECTIVE:To evaluate the clinical efficacy and safety of letrozole combined with traditional Chinese medicine for ovulation dysfunction induced by polycystic ovarian syndrome(PCOS). METHODS: 300 PCOS outpatients were randomized to 3 groups of 100 each: clomiphene citrate (CC) group (188 cycles);urinary gonadotropin (HMG) group (104 cycles) and letrozole (LE) group (155 cycles). The occurrence rate of mature follicle,ovulation rate,pregnancy rate,abortion rate,superfoetation-terata rate,cancellation rate,incidence of no dominant follicle forming and OHSS etc were compared between the two groups after treatment. RESULTS: 30 babies were born in CC group,34 in HMG group and 43 in LE group were born. The occurrence rate of mature follicles,ovulation rate and pregnancy rate in the LE group were higher than in either CC group or HMG group(P0.05);the superfoetation rate in HMG group was higher than in CC group or LE group(P

Potency is one of the most important indexes of inactivated vaccines.A number of methods have been established to assay the potency,of which the NIH test and single-dose mouse protection test are the prescribed methods.Here,we report a method to semi-quantitatively assay the potency of an inactivated rabies vaccine,which uses fewer animals and takes less time to complete.Depending on the quality requirements of a vaccine(e.g.minimum potency),a rabies reference vaccine is,for example,diluted to the minimum potency,and 50 μL of the dilution is taken to inoculate 10 mice.The same amount of the test rabies vaccine is inoculated into another 10 mice.After two weeks,all mice are bled and serum samples are assayed for viral neutralizing antibody by the fluorescent antibody virus neutralization(FAVN) test.By comparing the median and interquartile range of antibody titers of the reference vaccine with those of the test vaccine,the test vaccine potency can be semi-quantitatively judged as to whether it is in accord with the required quality.The reliability of this method was also confirmed in dogs.The procedure can be recommended for batch potency testing during inactivated rabies vaccine production.

BACKGROUND:Previous studies have mainly focused on costal cartilage, articular cartilage, nasal septal cartilage, and auricular cartilage, but in vitro culture of human vertebral endplate cartilage is stil rarely reported. OBJECTIVE: To discuss the feasibility of culture of vertebral endplate chondrocytes from type I neurofibromatosis associated with scoliosis patientsin vitro and to study the biological characters of the chondrocytes. METHODS: Through two-step enzymatic digestion and tissue culture, the chondrocytes from the vertebral endplate of seven type I neurofibromatosis patients isolated and cultured in monolayer and passaged to observe the changes of cel morphology under inverted phase contrast microscope. Colagen type II expression was detected by immunocytochemistry to identify whether the cels had chondrocyte characters. The growth kinetics was detected by using MTT colorimetric assay to draw the growth curve of passage 2 chondrocytes. RESULTS AND CONCLUSION:A few chondrocytes crawled from the cartilage after 2 weeks culture and cels were passaged at 3 weeks. Along with passage going on, the phenotype of chondrocytes was changed from polygonal, round, triangle, and irregular shapes to fusiform. The colagen type II expression in passage 2 cels was positive by immunohistochemical staining. MTT test showed the growth curve of the passage 2 chondrocytes presented a transverse “S”. Cels were found logarithmic growth at days 4-7, reached platform stage at days 8-13, and decreased at day 14. It is an effective and simple procedure by two-step enzymatic digestion and tissue explant method to culture vertebral endplate chondrocytes with high purity and good viability from type I neurofibromatosis patients associated with scoliosisin vitro. Passage 2 chondrocytes from the vertebral endplate exhibit the best viability at days 4-7, which can be used as targets for research of pathogenesis of type I neurofibromatosis with atrophic scoliosis.

Objective To investigate whether serum level of low-density lipoprotein cholesterol can predict the expan?sion of hemorrhage growth in elderly male patients with acute hypertensive intracerebral hemorrhage. Methods Patients (n=108) who visited our hospital with from June 2012 until May 2014 spontaneous hypertensive intracerebral hemorrhage with?in 6 hours of onset which is confirmed by initial computed tomography (CT) were sent to repeated CT within 24 hours of on?set. All selected patients were divided into the LDL-C≥2.49 mmol/L group and LDL-C<2.49 mmol/L group. Clinical data of these 2 groups were compared and the relationships of hematoma growth and its risk factors were analyzed. Results Baseline blood pressure, the level of blood glucose, PT, APTT, FIB, PLT and hemorrhage volume did not differ significantly between the LDL-C≥2.49 mmol/L group and LDL-C<2.49 mmol/L group. The ratio of hemorrhage growth in LDL-C<2.49 mmol/L group was significantly higher than that in LDL-C≥2.49 mmol/L group (34.21%vs 11.43%). Multiple logistic regres?sion analysis showed that LDL-C<2.49 mmol/L was the only risk factor contribute to hemorrhage growth. Conclusion Pa?tients with LDL-C<2.49 mmol/L in acute intracerebral hemorrhage are of high risk of hemorrhage growth so early attention and appropriate procedure are needed to prevent or slow its growth.

Objective To investigate the mechanism of restenosis after carotid stent and balloon angioplasty for the Guangxi swines by intravascular ultrasound(IVUS). Methods Twelve Guangxi swines fed by a high cholesterol diet were randomly divided into two groups. Seven stents were implanted in the left carotid artery of six swines in the first group, and balloon angioplasty was performed in the left carotid artery of swines in the other group. Digital subtraction angiography(DSA) and IVUS were conducted respectively before and after the intervention and in the 13th week. Results IVUS found that the percentage of area stenosis in stent group was (18.31±7.79) % and in balloon group (37.28±7.89) % in the 13th week. The percentage of area restenosis in stent was obviously related to neointimal hyperplasia (r = 0.897, P<0.05), the percentage of area restenosis due to balloon angioplasty was markedly related to area decrease of external elastic lamina (r = 0.856, P<0.05). Conclusions The restenosis in stent was related to intimal hyperplasia of blood vessel,and restenosis after balloon angioplasty had some connection with area decrease of external elastic lamina.

Objective To observe the safety and efficacy of the posterior spinal instrumentation (PSI) with preserving of the intraspinal rib head in neurologically intact patients with scoliosis secondary to Neurofibromatosis type 1 (NF1).Methods The clinical and radiographic data of nine NF1 scoliosis patients with rib head protrusion into the spinal canal,who had undergone PSI from August 1998 to March 2012,were retrospectively investigated.The average age of these patients (4 males,5 females) was 15.4±7.6 years,and their neurological status all were Frankel grade E preoperatively.The intraspinal rib head was not resected in all patients intraoperatively.The following parameters,including the magnitude of rib head penetration into the spinal canal (MRPC),the angle between the bilateral rib (ABR),the angle between the dislocated rib and the posterior vertebral wall (ARV),the coronal Cobb angle and the sagittal Cobb angle,were measured before and after surgery.Results The follow-up period was 0.5 to 4.8 years.The average MRPC decreased from preoperative 32.8％±9.9％ to postoperative 16.8％±15.2％ (P=0.026); the average ARV increased from preoperative 34.7°±16.4° to postoperative 47.8°±17.5° (P=0.001); the average ABR decreased from preoperative 83.0°±19.5° to postoperative 67.9°±13.3° (P=0.009); the average coronal Cobb angle decreased from preoperative 70.7° ±17.7° to 35.4°±17.0° immediately after operation (P=0.000) and the sagittal Cobb angle decreased from preoperative 59.7°±17.6° to 24.7°±10.8° immediately after operation (P=0.001).The coronal Cobb angle and sagittal Cobb angle had no significant change during follow-up period.The neurological status was Frankel grade E in all patients immediately after operation and at final follow-up.Conclusion For NF1 scoliosis patients with rib head penetration into the spinal canal without impingement of the spinal cord and neurological deficits,the deformity can be corrected safely and effectively without resecting the intraspinal rib head.

Objective To assess the efficacy of pirfenidone combined with PD-L1 inhibitor for treatment of bladder cancer in a mouse model and its effect on tumor immune microenvironment modulation.Methods Forty C57BL/6 mouse models bearing ectopic human bladder cancer xenografts were randomized into control group,PD-L1 inhibitor group,pirfenidone group and combined treatment group(n=10).After successful modeling,PD-L1 inhibitor treatment was administered via intraperitoneal injection at 12.5 mg/kg every 3 days,and oral pirfenidone(500 mg/kg)was given on a daily basis.The survival rate of the mice and tumor growth rate were compared among the 4 groups.The expressions of CD3,CD8,CD45,E-cadherin and N-cadherin in the tumor tissues were detected with immunohistochemistry after the 21-day treatment,and bone marrow-derived suppressor cells(MDSCs)were observed with immunofluorescence staining;serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),urea nitrogen(BUN),creatinine(CRE)and lactate dehydrogenase(LDH-L)were analyzed using an automated biochemical analyzer.Results Treatment with PD-L1 inhibitor and pirfenidone alone both significantly decreased tumor growth rate and tumor volume at 21 days(P<0.05),but the combined treatment produced an obviously stronger inhibitory effect(P<0.05).PD-L1 inhibitor and pirfenidone alone significantly increased E-cadherin expression and decreased N-cadherin expression in the tumor tissue(P<0.05).The two treatments both significantly increased the percentage of CD3+,CD8 and CD45+ T cells and decreased the percentage of Ly-6G+CD11b+MDSCs in the tumor tissue,and these changes were more obvious in the combined treatment group(P<0.05).No significant differences were found in serum ALT,AST,BUN,CRE or LDH-L levels among the 4 groups(P>0.05).Conclusion Combined treatment with pirfenidone and PD-L1 inhibitor significantly inhibits the progression of bladder cancer in mice possibly by regulating tumor immune microenvironment and inhibiting epithelial-mesenchymal transition of the tumor cells.

Objective To assess the efficacy of pirfenidone combined with PD-L1 inhibitor for treatment of bladder cancer in a mouse model and its effect on tumor immune microenvironment modulation.Methods Forty C57BL/6 mouse models bearing ectopic human bladder cancer xenografts were randomized into control group,PD-L1 inhibitor group,pirfenidone group and combined treatment group(n=10).After successful modeling,PD-L1 inhibitor treatment was administered via intraperitoneal injection at 12.5 mg/kg every 3 days,and oral pirfenidone(500 mg/kg)was given on a daily basis.The survival rate of the mice and tumor growth rate were compared among the 4 groups.The expressions of CD3,CD8,CD45,E-cadherin and N-cadherin in the tumor tissues were detected with immunohistochemistry after the 21-day treatment,and bone marrow-derived suppressor cells(MDSCs)were observed with immunofluorescence staining;serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),urea nitrogen(BUN),creatinine(CRE)and lactate dehydrogenase(LDH-L)were analyzed using an automated biochemical analyzer.Results Treatment with PD-L1 inhibitor and pirfenidone alone both significantly decreased tumor growth rate and tumor volume at 21 days(P<0.05),but the combined treatment produced an obviously stronger inhibitory effect(P<0.05).PD-L1 inhibitor and pirfenidone alone significantly increased E-cadherin expression and decreased N-cadherin expression in the tumor tissue(P<0.05).The two treatments both significantly increased the percentage of CD3+,CD8 and CD45+ T cells and decreased the percentage of Ly-6G+CD11b+MDSCs in the tumor tissue,and these changes were more obvious in the combined treatment group(P<0.05).No significant differences were found in serum ALT,AST,BUN,CRE or LDH-L levels among the 4 groups(P>0.05).Conclusion Combined treatment with pirfenidone and PD-L1 inhibitor significantly inhibits the progression of bladder cancer in mice possibly by regulating tumor immune microenvironment and inhibiting epithelial-mesenchymal transition of the tumor cells.

Objective To construct a RNA interference lentiviral vector aimed at human ARK5 (AMPK-related protein kinase 5) gene and explore its effect on the biologic behavior of human gastric cancer SGC7901 cells.Metho ds Targeting human ARK5 mRNA coding sequence, we designed three specific short hairpin RNAs (shRNAs) and constructed the lentiviral vector,then infected human gastric cancer SGC7901 cells with this vector.Afterwards, we used qPCR and Western blot for detecting the silencing effect on ARK5 gene,MTT colorimetric assay to measure the cell proliferation , cell scratch test for cell migration and Transwell for cell invasion, and flow cytometry analysis for apoptosis in cells treated with glucose starvation and TNF-α.Results Sequencing proved that the recombinant lentiviral vector containing ARK5-shRNA-3 was constructed successfully.Real time fluorescent quantitative PCR assay showed that the expression abundance of ARK5 gene in the normal control group, negative control group and ARK5-shRNA-3 infected group were 1.002+0.082, 1.001+0.050 and 0.140+0.003, respectively, showing a statistically significant difference (P<0 .01).Cell scratch test showed that the cell migration rate of ARK5-shRNA-3 infected group was (38.5+4.3 )%, significantly lower than that of the normal control group [(72.4+6.4)%] and negative control group [(75.1+7.1)%, P<0.01].The results of Transwell test showed that the number of penetrating cells in the normal control group, negative control group and ARK5-shRNA-3 transfection group were 257.4±12.3, 245.7±11.6, 112.5±7.8, with a significant difference (P<0.01).After glucose starvation and TNF-α-treatment for 24 h, the cell death rate of the normal control group, negative control group and ARK5-shRNA-3 group were (11.7±3.2)%, (12.3±2.6)% and (30.8±4.3)%, respectively, showing that the cell apoptosis rate of ARK5-shRNA-3 transfected group was significantly higher than that of the normal control and negative control groups (P<0.01).Conclusions We have successfully constructed a recombinant lentiviral vector which can efficiently silence ARK5 gene.Using it we can inhibit the proliferation, migration, invasion of tumor cells, and promote cell apoptosis under the condition of TNF-αtreatment and glucose starvation.