Astragalus Plant ; Cell Differentiation ; drug effects ; Cell Division ; drug effects ; Codonopsis ; Drugs, Chinese Herbal ; pharmacology ; Hematopoietic Stem Cells ; cytology ; drug effects

AIM: Currently available anti-scarring regimens for glaucoma filtration surgery have potentially blinding complications and safer alternatives would be beneficial. This experiment is to investigate the effect of TGF-β2 antisense oligodeoxynucleotide on differentiation, proliferation of subconjunctival fibroblast following glaucoma filtration surgery.METHODS: Glaucoma filtration surgery were performed on both eyes of 28 rabbits. TGF-β2 antisense oligodeoxynudeotide was subconjunctivally injected in the right eyes (A group), and TGF-β2 missense oligodeoxynucleotide (B group)or PBS(C group) was used at the same method in the left eyes as controls. Rabbits were killed at 4,7,14 and 28 days after surgery. Intraocular pressure (IOP), bleb characteristics were recorded at different time point. Subconjunctival fibroblasts were examined by immunohistochemistry and electron microscopy.RESULTS: The IOP of rabbits in group A was significantly lower at 14 days (6.74± 1.18 mmHg) and 21 days (8.15± 1.97mmHg) after operation than the IOP in group B (8.53± 1.04,9.72± 1.09 mmHg)(P ＜0.01) and group C(8.79± 1.21, 9.43±1.27 mmHg) (P ＜0.05). The mean bleb survival time was longer (17.2 days) in group A than that of group B (14.5 days) and group C (13.5 days)(P＜0.05). The population of the cells expressing α -smooth muscle actin(α -SMA) and proliferating cell nuclear antigen (PCNA) was significantly reduced in group A compared with the group B and C. The ultrastructure of fibroblast was not altered by TGF-β2 anti-sense oligodeoxynucleotide.CONCLUSION:TGF-β2 antisense oligodeoxynucleotide can prevent the scar formation after glaucoma surgery by inhibit the differentiation and proliferation of subconjunctival fibroblast. It could be a potentially useful anti-scarring alternative for the prevention of late surgical failure.

AIM:To compare the effect of 1,2-propanediol (PROH) and ethylene glycol (EG) on apoptosis and expressions of P53,Bcl-2 of follicles in human ovarian tissue,in order to offer experimental foundation for selecting the best cryoprotectant. METHODS:Biopsies of ovarian tissue obtained from 12 women were cryopreserved,and ovarian tissue slice from each woman was divided into three groups:fresh control group,ethylene glycol group and 1,2-propanediol group. The slow-freezing /rapid-thawing protocol was used to freeze and thaw the slice of ovarian cortex. Apoptosis of follicles in fresh and frozen-thawed ovarian cortical tissue was detected by TUNEL experiment,and expressions of P53 and Bcl-2 were detected by immuno-histochemistry. RESULTS:The percentages of apoptosis follicles were 14.58%,23.08%,30.43% in fresh control group,PROH group and EG group,respectively,and the percentage of apoptosis follicles in EG group was higher than that in fresh control group (P0.05). The percentages of P53 positive follicles were 13.48%,25.00% and 33.93%,respectively. There was significant difference between fresh control and EG group (P0.05). CONCLUSION:The results of this study indicate that 1.5 mol/L PROH is more suitable for cryopreservation of human ovarian tissue rather than 1.5 mol/L EG in slow-freezing /rapid-thawing protocol. The protocol of slow-freezing/ rapid-thawing may preserve oocytes well,but it is not ideal for cryopreservation of granulosa cells. Cryopreservation may influence on apoptosis of follicles in human ovarian tissue.

Objective To investigate the expression of Rap1 GTPase-activating protein 1 (Rap1GAP),matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9),and their relation with clinical patterns in colorectal carcinoma.Methods Immunohistochemistry was used to detect the expression of Rap1GAP,MMP-2 and MMP-9 in colorectal carcinoma,villous adenoma,tubular adenoma and normal colorectal tissue,and their relationship with clinicopathological parameters was analyzed.Results The positive rate of Rap1GAP expression was 30.4 ％ (14/46),77.8 ％ (14/18),69.6 ％ (16/23) and 95.2 ％ (20/21) in colorectal carcinoma,villous adenoma,tubular adenoma,and normal colorectal tissue,respectively (x2 =30.659,P=0.000).The figures were 71.7 ％ (33/46),55.6 ％ (10/18),52.2 ％ (12/16) and 9.5 ％ (2/21) for the positive rate of MMP-2 expression (x2 =22.459,P =0.000),as well as for 76.1％ (35/46),61.1％ (11/18),56.5 ％ (13/23) and 14.3 ％ (3/21) for the positive rate of MMP-9 expression,respectively (x2 =22.643,P =0.000).In patients with colorectal carcinoma,the expression of Rap1GAP was correlated with tumor differentiation (x2 =5.275,P =0.022),but not sex,age,or lymphatic metastasis (all P ＞ 0.05).The expression of MMP-2 and MMP-9 were correlated with lymphatic metastasis (x2 =6.661,P =0.010;x2 =8.475,P =0.040),but not sex,age or tumor differentiation(all P ＞ 0.05).There was a negative correlation between expression of Rap1GAP and MMP-2,MMP-9 in colorectal carcinoma,respectively (r =-0.424,P =0.003;r =-0.294,P =0.048),but no correlation between the expression of MMP-2 and MMP-9 (r =0.101,P =0.505).Conclusions Rap1GAP,MMP-2 and MMP-9 play important roles in the malignant biological behavior of colorectal carcinoma,and the expression of Rap1GAP is negatively correlated with MMP-2 and MMP-9.The interactions among the three affect the occurrence and development of colorectal carcinoma.

Objective To investigate the clinicopathological significance of periostin and MMP-2 in colorectal carcinoma.Methods Immunohistochemistry was used to detect the expression of periostin and MMP-2 in colorectal carcinoma,tubular adenoma,villous adenoma and normal colorectal tissue.The correlations between the expression of periostin and MMP-2 with clinicopathologic parameters were analyzed.Results In colorectal carcinoma,tubular adenoma,villous adenoma and normal colorectal tissue,the positive rates of periostin were 83.7 ％ (41/49),40.0 ％ (6/15),32.1％ (9/28),0 (0/15),respectively,while the positive rates of MMP-2 were 71.4 ％ (35/49),60.0 ％ (9/15),64.3 ％ (18/28),0(0/15),respectively.There were significant differences among the groups (x2 =41.252,P =0.000; x2 =24.811,P =0.000).The expression of periostin in colorectal carcinoma tissue were not correlated with sex (x2 =0.002,P =0.961),age (x2 =2.267,P =0.132),tumor sites (x2 =1.506,P =0.220),differentiation status (x2 =0.875,P =0.350) and lymphatic metastasis (x2 =3.315,P =0.069).The expression of MMP-2 in colorectal carcinoma were correlated with lymphatic metastasis (x2 =5.800,P =0.016),whereas not correlated with sex (x2 =0.562,P =0.453),age (x2 =0.138,P =0.711),tumor sites (x2 =0.408,P =0.532),differentiation degree (x2 =1.335,P =0.248).The expression of periostin and MMP-2 were positively correlated in colorectal carcinoma (r =0.332,P =0.020).Conclusion Periostin and MMP-2 are correlated closely with the development and progression of colorectal carcinoma.It might be helpful for evaluating the biological properties and prognosis of colorectal carcinoma,and it would be more accurate to use a combined analysis of the two indicators.

iple occurrence of the tumor, which can decline its recurrence and postpone its progression.

The angiotensinogen(AGT) expression and angiotensin Ⅱ (AngⅡ ) secretion levels in cultured SD rat mesangial cells were determined. High glucose up-regulated AGT mRNA(0. 29±0.07 vs 0. 20±0. 05,P< 0.05)and protein(0.66±0.23 vs 0.37±0. 15,P<0.05) expression and Ang Ⅱ secretion [(9.85±2.08 vs 7.50± 1. 51) pg/ml,P<0. 05]levels, which were down-regulated by pyrrolidine dithiocarbamate( PDTC) treatment via inhibiting NF-κB activity.

Objective To observe the effects of the compatibility of Radix Astragali and Radix Angelicae Sinensis in different proportion on hemodynamic and balance of Th1/Th2 cells of Qi deficiency and blood stasis rat model. Methods Qi deficiency and blood stasis rats were caused by restricted diet, forced swimming and norepinephrine subcutaneous injecting, and treated by compatibility of Radix Astragali and Radix Angelicae Sinensis in different proportion of 5∶1, 3∶1, 1∶1 and 1∶2 for 21 days. The indexes of hemorheology were detected with hemorheological analyser, and the level of IFN-γand IL-4 in serum were tested by ELISA. Results Compared with model group, the compatibility of Radix Astragali and Radix Angelicae Sinensis in proportion of 1∶1, 3∶1 and 5∶1 groups reduced the low shear blood viscosity. The spleen index of model group decreased, and compatibility of Radix Astragali and Radix Angelicae Sinensis in 5∶1 proportion group increased spleen index. The level of IL-4 increased and IFN-γ decreased in the serum of model group, the compatibility of Radix Astragali and Radix Angelicae Sinensis in proportion of 3∶1 group increased the level of IFN-γ. Astragalus angelica 5∶1 group decreased the level of IL-4 and increased the level of IFN-γ. Conclusion The compatibility of Radix Astragali and Radix Angelicae Sinensis can improve hemorheology, adjust the balance of Th1/Th2 cells of Qi deficiency and blood stasis rats. The effects were better when Radix Astragali’s dosage greater than that of Radix Angelicae Sinensis, and the group of 5∶1 proportion was the best.

Objective To establish the quality standards of Longdan Xiegan Capsule and Longdan Xiegan Soft Capsule. Methods Gentianae Radix et Rhizoma, Bupleuri Radix, Scutellariae Radix, Gardeniae Fructus, Alismatis Rhizoma, Angelicae Sinensis Radix, Rehmanniae Radix and Glycyrrhizae Radix et Rhizoma were identified by TLC. Gentiopicrin in Gentianae Radix et Rhizoma, geniposide in Gardeniae Fructus, baicalin in Scutellariae Radix were determined by HPLC. Results The TLC spots developed were clear. Gentiopicrin showed good linear relationship in the range of 0.066 1-0.595 1 mg (r=1.000 0), the average recovery of Capsule was 99.34%(RSD=1.50%), and the average recovery of Soft Capsule was 96.62%(RSD=1.50%). Geniposide showed good linear relationship in the range of 0.076 1-1.369 8 mg (r=0.999 9), the average recovery of Capsule was 101.3% (RSD=1.70%), and the average recovery of Soft Capsule was 100.59%(RSD=0.79%). Baicalin showed good linear relationship in the range of 0.214 4-1.608 mg (r=1.000 0), the average recovery of Capsule was 101.12% (RSD=1.30%), and the average recovery of Soft Capsule was 98.07% (RSD=2.40%). Conclusion The method is simple and reproducible. It can be used to control the quality of Longdan Xiegan Capsule and Longdan Xiegan Soft Capsule.

Objective To explore the methylation status of Rap1 GTPase activating protein (Rap1GAP) promoter in colon cancer, and to provide the oretical basis and research direction for the early diagnosis, targeted therapy, anti-multidrug resistance of colon cancer and so on. Methods The paraffin embedded specimens of 33 patients with colonic adenocarcinoma diagnosed by pathology were analyzed from Department of Pathology of Xinzhou City People′s Hospital from January 2010 to September 2014, including 19 males and 14 females, and aged 41-72 years old. The paraffin embedded specimens of 16 patients with colonic adenoma were enrolled, including 9 males and 7 females, and aged 34-58 years old. 13 normal tissues from the tumor distal margin (from the tumor > 15 cm) were selected. Quantitative methylation specific PCR (q-MSP) was applied to detect methylation level of Rap1GAP gene promoter. The methylation level differences of Rap1GAP gene promoter region among 3 groups or between different clinicopathologic factor subgroups were compared. Results The methylation rates [median (interquartile range)] of Rap1GAP promoter were 65.43 % (50.35 %), 21.37 % (8.39 %) and 17.43 % (15.71 %) in colonic adenocarcinoma group, colonic adenoma group and adjacent normal tissue group, respectively. The methylation rate of colonic adenocarcinoma group was significantly higher than that of colon adenoma group or that of adjacent normal tissue group (P< 0.05). The methylation rates of Rap1GAP promoter in colonic adenocarcinoma were not correlation with age, sex, differentiation and the stage of TNM [ male vs. female: 42.74 % (70.44 %) vs. 21.98%(80.00%);≤60yearsoldvs.>60yearsold:36.26%(62.62%)and26.23%(76.42 %);well-differentiated vs. moderately/poorly-differentiated: 21.98 % (40.32 %) vs. 42.74 % (74.20 %); TNM Ⅰ-Ⅱ vsⅢ-Ⅳ: 25.31 % (48.27 %) vs. 36.26 % (75.55 %); all P> 0.05]. Conclusion The methylation status of RAP1GAP promoter maybe associate with genesis and development of colon cancer, which might be used as a target for early diagnose of colon cancer.