[Objective]This study was aimed to research the synergistic antitumor proliferation effects and their best proportion of ursolic acid(UA) and tetrandrine(Tet), a pair of compounds isolated from Chinese herbs which showed complement inhibition on the multiple signal pathways. [Methods] The reporter assays on tumor-related signal pathways for MAPK/ERK, MAPK/JNK, NF-κB, Wnt, Notch, Cell Cycle, Myc/Max and Hypoxia were used to study the effect of five different Chinese herbal compounds on tumor proliferation,it was concluded cepharanthine(Cep), Tet, 18α-glycyrrhetinic acid(18α-Gly), UA and luteolin(Lut). MTT assay and crystal violet staining were used to study the antiproliferative effect of 15 different compounds for the tumor cells of MDA-MB-231,SW480,MG63,PC3,DU145,HCT116,143B and MDA-MB-468, which is consisted with Cep, Tet, 18α-Gly, UA and Lut for the 15 different compounds. Coefficient of drug interaction(CDI) method was used to detect the synergistic effect of the two compounds. Combination of index(CI) and isobologram method was used to screen the best ratio of compounds in their antiproliferative effects. [Results] The signal pathway reporter assay showed that UA and Tet could complementarily inhibit tumor-related signaling pathways. And the results also showed that UA and Tet could induce synergetic anti-tumor cell proliferation in vitro. Furthermore, the optimal ratio of UA and Tet was 9:1 by using isobologram and CI method. [Conclusion] UA and Tet can be inhibited and complemented by 8 tumor-related signaling pathways, and we used MTT assay and crystal violet staining or other methods to confirm the synergistic antitumor proliferation effects, furthermore, the optimal proportion for UA and Tet were screened, and it provided a new insight to develop new anticancer formula in research.

This study was aimed to optimize the extraction process of resveratrol by central composite design (CCD) and response surface method (RSM), based on the result of single factor experiment. Resveratrol was extracted from Polygonum cuspidatum by water bath extraction of organic solvent. And content of resveratrol was taken as index in the investigation of solvent concentration effect, extraction time and solvent content on extraction process. The results showed that the optimum condition was 12.7 times amount of 72.0% ethanol and extract for 2 times under the temperature of 50℃, 1.1 h for each time; the average deviation between the maximum theoretical value and measured value was 0.83%. It was concluded that this extraction process was highly predictive, which provided experimental evidence for the industrial production of P. cuspidatum extraction.

Objective To explore the effect of transcription factor SOX18 on proliferation, migration and invasion of HeLa cells and its related mechanism. Methods HeLa cells were divided into blank control group, negative control group, pCI-SOX18 group. The expression of SOX18 and matrix metalloproteinase 7 (MMP-7) was detected by Western blot. The cell proliferation of different group was measured by MTT analysis. The effect of transcription factor SOX18 on migration and invasion of HeLa cells were measured by Transwell assay. Results HeLa cells were successfully transfected with pCI-SOX18. According to the results of MTT analysis, the cell number of pCI-SOX18 group showed no significant difference compared with blank control group and negative control group (P> 0.05). Compared to blank control group (0.23 ±0.05) and negative control group (0.28±0.07), SOX18 expression in pCI-SOX18 group (1.16±0.13) was significantly higher (F=218.14, P<0.05). Meanwhile, MMP-7 expression in pCI-SOX18 group (0.95±0.09) was increased significantly compared with blank control group (0.71±0.07) and negative control group (0.68±0.06) (F=116.84, P < 0.05). The number of cell migration in pCI-SOX18 group (175.71 ±17.17) was increased significantly compared with that in negative control group (95.19 ±8.18) and that in blank control group (93.34±9.72) (t=75.47, t=77.35, P<0.05). Compared with that in negative control group (98.63±14.62) and that in blank control group (96.57 ±13.95), the number of cell invasion in pCI-SOX18 group (247.12 ±36.82) was significantly higher (t=98.02, t=99.34, P<0.05). Conclusion Overexpression of SOX18 has no effect on the proliferation of HeLa cells, but has a significant role in promoting cell migration and invasion probably by regulating MMP-7 expression positively.

Objectives:In the study, effect of Ala Gln on glutathione concentration and antioxidation were investigated in burned rats . Methods:Thirty male SD rats were randomly divided into three groups,A:control group,B:TPN group,C:Ala Gln PN group.The nutrient solution was administered through central venous catheter for 7 days.Glutathione concentration and levels of xanthine oxidase(XOD) and superoxide dismutase(SOD) in the serum were determined. Results:Compared with control group and TPN group,the levels of serum glutathione and SOD in group C were significantly increased( P

Objective To design and investigate the clinical effects of parallel double venous pots hemodialysis tubing set in patients with heparin-free hemodialysis.Methods Totally 120 patients who received hemodialysis therapy in our hospital from May,2014 to June,2016 were randomly divided into the experimental group (n=60) and the control group(n=60) via random number table.The experimental group received self-designed parallel double venous pots dialysis tubing set,while the control group received the normal dialysis tubing set.The number of preventive replacement of dialyzer and tubing set during heparin-free dialysis,the number of discarded tubing set due to coagulation and the time frame for the first tubing set were recoded from two groups.Results There were statistically significant differences in the number of discarded tubing set due to coagulation and the time frame for the first tubing between two groups(P＜0.05).Conclusion Parallel double venous pots dialysis tubing set can reduce the number of discarded tubing set due to coagulation,prolong the time frame for the first tubing.

Objective To investigate clinical efficacy and safety and complications of transurethral plasmakinetic resection of prostate ( PKRP) for benign prostatic hyperplasia ( BPH) .Methods Totally 186 BPH patients were underwent PKRP .Comparison of clinical parameters before and after operation .Results Following-up at 3 and 6 months after the operation showed that international prostate symptom score ( IP-SS),quality of life(QQL),residual urine volume(RUV) scores increased and maximal urinary flow rate ( Qmax) scores decreased .The incidence of complications was 8.2%.Conclusion PKRP have efficacy in the treatment of BPH , and PKRP is safer and less complications .

Objective To study the treatment of acute renal failure induced by uretericobstruction . Methods Twenty-five cases of acute renal failure induced by uretericobstruction were emergent managed with ureteroscope pneumatic lithotripsy .Results All the renal function resumed well , BUN,Cr in serum was natural or near natural .Conclusion Ureteroscope pneumatic lithotripsy should be used for acute renal failure induced by uretericobstruction as first-line.

Infectious clone is a useful tool in exploring viral replication and pathogenesis. In order to prevent linear PCV2 cyclization, PCR mutagenesis was used to construct the first molecular clone (pSK-2PCV2) by ligating two copies of the complete PCV2 genome with the pBluescript SK (pSK) vector. In addition, pSK-PCV2 and ds-PCV2 were constructed. PK-15 cells were transfected with above three infectious clones. Indirect immunofluorescence assay (IFA) revealed that the virus antigen mainly localized in infected cell nucleolus and cytoplasm. PCV2 specific nucleotide fragment in cell culture was amplified by RT-PCR. Typical porcine circovirus particles with diameter about 17 nm were also observed by transmission electron microscope (TEM) in the infected cells. The rescued virus sequences from the cultures had 100% homology with the inserting PCV2 genome. The rescued virus shared similar properties with that of the parental virus. The study establishes a platform for further research on the virus molecular biology and pathogenicity.
Animals ; Cell Line ; Circoviridae Infections ; virology ; Circovirus ; genetics ; growth & development ; pathogenicity ; physiology ; Cloning, Molecular ; DNA, Viral ; genetics ; physiology ; Recombination, Genetic ; genetics ; Swine ; Transfection ; Virulence ; Virus Replication

Objective:To evaluate the role of Toll-like receptor 4 (TLR4)/nuclear transcription factor κB (NF-κB) signaling pathway in long-term cognitive impairment induced by multiple exposures to sevoflurane anesthesia in neonatal rats.Methods:Seventy-five SPF healthy newborn Sprague-Dawley rats of either sex, aged 6 days, weighing 12-20 g, were divided into 3 groups ( n=25 each) using a random number table method: control group (group C), multiple exposures to sevoflurane anesthesia group (group S) and TLR4 inhibitor plus multiple exposures to sevoflurane anesthesia group (group I+ S). The rats in group S and group I inhaled 3% sevoflurane for 2 h at 6, 7 and 8 days after birth. TLR4 inhibitor TAK-242 10 mg/kg was intraperitoneally injected before each exposure to sevoflurane in group I, and the equal volume of normal saline was given instead in the other two groups. The spontaneous activity was evaluated by open field test on day 29 after birth, and the cognitive function was assessed by Morris water maze test on days 30-34 after birth. After the behavioral test, the blood samples from the abdominal aorta were collected, and then the rats were sacrificed under deep anesthesia to isolate the hippocampal tissues for measurement of the levels of S100β and neuron-specific enolase (NSE) in serum and hippocampal interleukin-1β (IL-1β), IL-6 and tumor necrosis factor α (TNF-α) (by enzyme-linked immunosorbent assay), expression of TLR4, NF-κB p65 and phosphorylated NF-κB p65 (p-NF-κB p65) (by Western blot) and for microscopic examination of the pathological changes of hippocampal CA1 region after HE staining. Results:Compared with group C, the escape latency was significantly prolonged, the number of crossing the original platform was reduced, the TLR4 expression was up-regulated, the ratio of p-NF-κB p65/NF-κB p65 was increased, the levels of serum S100β protein and NSE and hippocampal IL-1β, IL-6 and TNF-α were increased ( P<0.05), and the pathological changes in the hippocampal CA1 region were aggravated in group S. Compared with group S, the escape latency was significantly shortened, the number of crossing the original platform was increased, TLR4 expression was down-regulated, the ratio of p-NF-κB p65/NF-κB p65 was decreased, the levels of S100β and NSE in serum and hippocampal IL-1β, IL-6 and TNF-α were decreased ( P<0.05), and the pathological changes in hippocampal CA1 area were significantly attenuated in group P. Conclusions:The mechanism by which multiple exposures to sevoflurane anesthesia induces long-term cognitive impairment is related to activation of TLR4/NF-κB signaling pathway and increase in hippocampal inflammatory responses in neonatal rats.

Objective:To evaluate the role of nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway in edaravone-induced attenuation of long-term cognitive impairment caused by long-time sedation with propofol in the neonatal rats.Methods:Eighty SPF healthy newborn Sprague-Dawley rats of both sexes, aged 7 days, weighing 15-20 g, were divided into 4 groups ( n=20 each) using a random number table method: control group (group C), propofol group (group P), edaravone+ propofol group (group EP) and Nrf2 inhibitor ML385+ edaravone+ propofol group (group MEP). Propofol 75 mg/kg was intraperitoneally injected once a day for 7 consecutive days in P group, EP group and MEP group, respectively, while the equal volume of medium/long chain fat emulsion injection was intraperitoneally injected in C group. Edaravone 3 mg/kg was intraperitoneally injected at 30 min before each propofol injection in EP and MEP groups, and ML385 15 mg/kg was intraperitoneally injected simultaneously in group MEP. The spontaneous activity was evaluated by the open field test on day 29 after birth, and the cognitive function was assessed by Morris water maze test on days 30-34 after birth. The rats were sacrificed after the end of water maze test, and brains were removed and hippocampal tissues were obtained for determination of reactive oxygen species (ROS) levels (by flow cytometry), superoxide dismutase (SOD) and malondialdehyde (MDA) levels (by enzyme-linked immunosorbent assay) and expression of Nrf2 and HO-1 (by Western blot) and for microscopic examination of the pathological changes in the hippocampal CA1 area (using HE staining). Results:There was no significant difference in the speed, distance and time of stay at the center of the open field among the four groups ( P>0.05). Compared with C group, the escape latency was significantly prolonged, the number of crossing the original platform quadrant was reduced, the levels of MDA and ROS were increased, the activity of SOD was decreased, the expression of Nrf2 and HO-1 was down-regulated ( P<0.05), and the pathological injury was observed in the hippocampal CA1 region in group P. Compared with P group, the escape latency was significantly shortened, the number of crossing the original platform quadrant was increased, the levels of MDA and ROS in the hippocampus were decreased, the activity of SOD was increased, the expression of Nrf2 and HO-1 was up-regulated ( P<0.05), and the pathological injury in the hippocampal CA1 region was significantly alleviated in EP group. Compared with EP group, the escape latency was significantly prolonged, the number of crossing the original platform quadrant was reduced, the levels of MDA and ROS were increased, the activity of SOD was decreased, the expression of Nrf2 and HO-1 was down-regulated ( P<0.05), and the pathological injury was aggravated in the hippocampal CA1 region in MEP group. Conclusions:The mechanism by which edaravone attenuates long-term cognitive impairment caused by long-time sedation with propofol is related to activation of Nrf2/HO-1 signaling pathway and inhibition of oxidative stress in the neonatal rats.