Objective To explore a reliable method of 70% hepatectomy model in liver fibrosis mice. Methods Sixty-six C57BL6 mice were randomly devided into control group (n=6), the traditional group (n=30, ligation and removal liver lobe) and improved group (n=30, removal of liver lobe after blocking blood flow). Those 60 mice were induced liver fibrosis firstly, then randomly divided into six mice in each group, and were sacrificed at preoperative, 12, 24, 48 and 72 hours after liver resection. Liver tissues and blood samples were collected. The survival rate and incidence of complications were recorded and compared between two groups. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured to observe the liver injury after 70%hepatectomy. The ratio of liver weight to body weight and the expression of proliferating cell nuclear antigen (PCNA) were also measured to observe the difference of liver regeneration between the two groups. Results (1) Compared to the pathological control group, liver fibrosis model was established successfully in both traditional group and improved group, which can be used in 70%hepatectomy. So the follow-up experiment can be undertook timely. (2) Compared to traditional group, the survival rate was improved significantly in improved group (96.67%vs. 73.33%), and the incidence of complications was significantly lower (P<0.05). (3) The ALT and AST levels were higher 12 h and 24 h after operation in traditional group than those of improved group (P<0.05), while ALT and AST levels were increased first 12 h after operation and then decreased in both groups (P<0.05). (4) The liver/body weight ratio showed a decreasing trend 12 h after hepatectomy in two groups. The expression of PCNA increased at the beginning of postoperative, and reached its peak at 48 h (P<0.05). However, there was no significant difference at each time point between the two groups. Conclusion By blocking blood flow to establish 70% hepatectomy model in liver fibrosis mice, we can significantly improve the success rate of the model, and reduce the incidence of complications.

Objective To evaluate effects of human papilloma virus(HPV)16E7 on expressions of six tumor suppressor genes(including MT1G, NMES1, RRAD, SFRP1, SPARC and TFPI2)in a cell line SiHa, as well as on its proliferation and apoptosis. Methods SiHa cells were divided into two groups to be transfected with a small interfering RNA targeting HPV16E7(E7SiRNA, experimental group)and an empty vehicle(negative control group) respectively, with SiHa cells receiving no treatment serving as the blank control group. After 48 hours, qPCR was performed to measure the mRNA expressions of E7 and six tumor suppressor genes, CCK?8 assay to evaluate cellular proliferative activity, and flow cytometry to assess apoptosis of SiHa cells. Results At 48 hours after the transfection, the experimental group showed significantly decreased E7 mRNA expression(0.25 ± 0.036, P<0.05), but increased mRNA expressions of the six genes(MT1G 1.403 ± 0.190, NMES1 1.720 ± 0.060, RRAD 1.390 ± 0.160, SFRP1 1.493 ± 0.120, SPARC 2.157 ± 0.144, TFPI2 2.060 ± 0.122, all P < 0.05). The proliferative activity of SiHa cells was significantly decreased(0.554 ± 0.130 vs. 1.028 ± 0.236 and 1.220 ± 0.126, both P<0.05), but the apoptosis rate was significantly increased(9.222%vs. 0.246%and 0.123%, both P<0.05)in the experimental group compared with the negative control group and blank control group. No significant differences were observed between the negative control group and blank control group in proliferative activity or apoptosis rate of SiHa cells(both P>0.05). Conclusion E7 may participate in HPV16?induced cellular malignant transformation by suppressing the mRNA expressions of 6 tumor suppressor genes, including MT1G, NMES1, RRAD, SFRP1, SPAR and TFPI2.

Objective To evaluate the safety and effectiveness of 2μm Thulium laser vaporization enucleation (Thu-VEP) in the treatment of benign prostate hyperplasia (BPH). Methods A total of 145 patients with BPH were randomized into two groups including patients underwent ThuVEP and patients underwent standard transurethral resection of the pros-tate (TURP). The intraoperative blood loss,flushing fluid quantity,operation time,bladder irrigating time,catheterization time,the international prostate symptom score (IPSS),quality of life score (QOL),the maximum flow rate (Qmax),the post-void residual (PVR) and complications were observed in two groups. Results There were no significant differences in pa-tient age, preoperative duration, prostate weight, IPSS score,QOL score,Qmax and RUV between two groups. Patients in two groups were performed surgery successfully. The values of blood loss, bladder irrigating time and catheterization time were significantly less in ThuVEP group than those of TUEP group (P＜0.05). There were no postoperative complications in pa-tients of ThuVEP group. There were 5 cases of complications in TUEP group after operation. The values of IPSS score,QOL score,Qmax and RUV were significantly different after 3-12-month follow up than those before operation(P＜0.01),but no significant difference between two groups. Conclusion ThuVEP is a safe and more effective treatment than that of TURP for patients with BPH. ThuVEP can significantly improve the quality of life, and reduce complications in patients with BPH.

Objective To study the promoting effects and mechanisms of interleukin-22 on liver regeneration in GCl4-induced liver fibrosis mice after partial hepatectomy.Methods One hundred and fortyfour C57/BL6 mice were randomly divided into four groups:PHX group,CCl4 group,CCl4 + PHX group,and CCl4 + IL22 + PHX group.The blood samples were taken to measure serum ALT and AST levels.ALT /AST was calculated to observe the liver injury at 3 h,6 h,12 h,24 h,48 h and 72 h after hepatectomy.The liver tissue specimens were collected at each time point after hepatectomy.We measured the hepatic lobe to calculate the liver weight ratio and conducted pathological examinations to observe the degree of fibrosis and pathological changes at each time point.The positive expression of proliferating cell nuclear antigen (PCNA) in liver tissue was tested by immunohistochemistry.The level of CyclinD1 and STAT3 (Signal transducer and activator of transcription 3) signaling pathway was detected by Western blot.Results (1) Compared with CCl4 + PHX group,the ALT/AST ratio of CCl4 + IL22 + PHX group was significantly higher at 24 h,48 h and 72 h,and the level of ALB of CCl4 + IL22 + PHX group was obviously increased at 48 h and 72 h (P ＜ 0.05).(2) The liver regeneration was significantly increased in CCl4 + IL22 + PHX group.Compared with CCl4 + PHX group (2.08 ± 0.16,2.77 ± 0.07,2.97 ± 0.14),the liver weight ratio of CCl4 + IL22 + PHX group(2.34 ± 0.07,3.23 ± 0.09,3.55 ± 0.09) dramatically increased at 24 h,48 h and 72 h.Moreover,the pathological sections displayed that the disease was alleviated (P ＜ 0.05).(3) Immunohistochemical assay and western blot revealed that compared with other three groups,the level of PCNA,STAT3 and Cyclin D1 was significantly lower in the CCl4 + PHX group.However,the level of PCNA,STAT3 and Cyclin D1 apparently increased in CCl4 + IL22 + PHX group at 24 h,48 h and 72 h (P ＜ 0.05).Conclusion Interleukin-22 may significantly promote liver regeneration and reduce liver pathological injury in liver fibrosis mice induced by administration of CCl4 after hepatectomy,which plays a positive role in the recovery of liver function.

AIM:To compare the effect of three types of pterygium surgery and on tear film in patients with type 2 diabetes mellitus. ·METHODS:A total of 102 patients ( 102 eyes ) with pterygium combined with type 2 diabetes mellitus treated in our hospital from March 2013 to March 2016 were analyzed retrospectively. The patients were divided into three groups including the 34 cases ( 34 eyes ) with simple excision of pterygium ( resection group ) , pterygium excision combined with conjunctival flap transplantation in 34 cases (34 eyes, as conjunctival flap group ) and pterygium excision combined with limbal stem cell transplantation in 34 cases ( 34 eyes, as stem cell group ) . The wound repair time, complications, recurrence rate, uncorrected visual acuity (UCVA), tear film break-up time ( BUT ) and basal tear secretion test (SⅠt) were observed before, and 6 and 12mo after surgery in the three groups, respectively. ·RESULTS:The postoperative UCVA of the three groups was significantly higher than that preoperation ( P =0. 039, 0. 013, 0. 024 ), and there was no significant difference among the three groups ( P = 0. 317 ). The wound repair time was 5. 67 ± 1. 45d in the resection group, which was significantly higher than that in the conjunctival flap group (4. 18 ± 0. 76d) and the stem cell group (4. 09±0. 79 d) (P<0. 001), there was no significant difference between the conjunctival flap group and the stem cell group ( P = 0. 937 ). There were 4 cases in resection group reappeared, and the recurrence rate was 11. 8%, which was significantly higher than the other two groups ( P = 0. 037 ). There were 1 recurrences in the conjunctival flap group, and the recurrence rate was 2. 9%, while the patients in the stem cell group had no obvious recurrence. SⅠt and BUT increased significantly after operation (P<0. 05), especially in conjunctival flap group and stem cell group (P<0. 001). There was no significant difference between the conjunctival flap group and the stem cell group (P=0. 845, 0. 894). · CONCLUSION: Pterygium excision combined with conjunctival flap transplantation or limbal stem cell transplantation for the treatment of type 2 diabetic patients with normal blood glucose and tear film function has the similar effect, and is better than simple pterygium excision.

Attention was gradually paid by biologists to the using of RNA secondary structure in the classification of microbiology and phylogenetic relationship analysis in recent years. The development around the research was summarized here briefly. And more emphasis was given to the part introducing the application of RNA secondary structure to the analysis of phylogenetic relationship.

Eighty-five endophytic fungal strains were isolated from the roots、stems and leaves of Polygala tenuifolia Willd, among which, fifty-two from natural plants and thirty-three from cultivated ones. Sev-enty-six strains were classified as twenty-three fungal genera. The inhibitory activity screening to fourteen microbe were conducted research. The results showed that some endophytic fungi had remarkble inhibitory activities to Bacillus subtilis, Shigella sonnei, Escherichia coli, Candida albicans, Fusarium kyrushuense and they were all belonged to Fusarium, Alternaria, Aphanocladium respectively. All of the endophytic fungi isolated from Polygala tenuifolia showed no inhibitory activities to Staphyloccocus aureus, Salmonel-lae enteritis, Bibrio parahemolyticus.

Objective To screen for the miR-155 expression in synovial fibroblasts of rheumatoid arthritis (RASFs) and osteoarthritis (OASFs) and to evaluate the function of miR-155 on RASFs and its possible target mRNAs. Methods The expression levels of miR-155 in RASFs and OASFs were detected by real-time PCR. MiR-155 mimic and miR-155 inhibitor, as well as scrambled control were transfected into cultured RASFs by Lipofectamine 2000. Forty-eight hours later, MMP-3 levels in the cell culture supernatant were detected by ELISA and fibroblast proliferation was assayed by 3H -TdR incorporation test. Fibroblast invasive ability was tested by transwell system. IKBKE which previously identified as actual target of miR-155 was examined by real-time PCR. Comparisons between groups were performed with t test or one-way ANOVA analysis. Results It was shown that miR-155 was up-regulated in RASFs (1.79 ±1.94) and it was higher than that in OASFst (0.11±0.17), P<0.05]. Up-regulation of miR-155 could decrease MMP-3 levels (P<0.05). The proliferation and invasion of RASFs transfected with miR-155 were both evidently suppressed (P<0.05), while reducing the endogenous miR-155 could significantly enhance RASF proliferation (P<0.05). The expression of IKBKE of RASFs transfected with miR-155 was obviously down-regulated compared to those transfected with the scrambled control (P<0.05). Conclusion miR-155 is up-regulated in RASFs which may be a protective factor against the inflammatory effect, at least partially by attenuating the expression of IKBKK.

To explore the new method of discriminating Cistanche deserticola, Cynomorium songaricum and Orobanche pycnostachya by using PCR amplification of specific alleles. 30 samples of the different C. deserticola, 21 samples of C. songaricum and O. pycnostachya were collected. The total DNA of the samples were extracted, the ITS sequences from C. deserticola, C. songaricum and O. pycnostachya were amplified by PCR and sequenced unidirectionally. These sequences were aligned by using ClustulW. Specific primer was designed according to the ITS sequences of specific alleles, and PCR reaction system was optimized. Additionally, compare with the identification of specific PCR method and DNA sequence analysis method. The result showed that the 331 bp identification band for C. deserticola and the adulterants not amplified bands by a single PCR reaction, which showed good identification ability to the three species. PCR amplification of specific alleles can be used to identify C. deserticola, C. songaricum and O. pycnostachya successfully.
Alleles ; Cistanche ; classification ; genetics ; DNA Primers ; genetics ; DNA, Intergenic ; genetics ; DNA, Plant ; genetics ; Drug Contamination ; prevention & control ; Phylogeny ; Polymerase Chain Reaction ; methods

Adjuvants, Immunologic ; therapeutic use ; Adult ; Aged ; B-Lymphocytes ; immunology ; CD4-CD8 Ratio ; Carcinoma, Hepatocellular ; drug therapy ; immunology ; surgery ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Killer Cells, Natural ; immunology ; Liver Neoplasms ; drug therapy ; immunology ; surgery ; Male ; Middle Aged ; Perioperative Care ; Phytotherapy ; T-Lymphocyte Subsets ; immunology