AIM: To investigate the influence of He-Ne laser on connective tissue growth factor (CTGF) expression and collagen formation of fibroblast in filtration site after trabeculectomy in rabbit, and to discuss the mechanism for preventing scar formation with He-Ne laserin vivo.METHODS: The upper nasal limbus area next to the upper rectus muscle in right eyes Received 10 minutes He-Ne laser irradiation (200mW/cm2) every day for three days, the left eyes served as control. Twenty-four hours after the last irradiation, both eyes of the rabbits were took trabeculectomy surgery. The expressions of CTGF in the filtration area were tested on the 7th, 14th and 28th day after surgery and collagen density was tested on the 14th and 28th day after surgery. Each of the time point had 7 rabbits. RESULTS: The expression of CTGF was lower than that of the control group's on the 7th and 14th day after trabeculectomy surgery (P=0.01, P=0.005). When examined on the 14th and 28th day, the collagen density of irradiation group were significantly lower than that of the control group's (P=0.013, P=0.01).CONCLUSION: Pretreating the filtration area with 200mW/cm2 He-Ne laser may be helpful in preventing scar formation after trabeculectomy in rabbit, possibly due to downregulation of the expression of CTGF and collagen synthesis in fibroblasts. He-Ne laser may be developed into a new scar preventing method in filtration surgery.

Peptide drugs exhibit an irreplaceable role in clinics due to their high specificity, efficiency and low toxicity. At present, more than 80 peptide drugs have been approved for marketing with global sales exceeding $50 billion in 2019. However, with large molecular weights, high hydrophilicity and instability in digestive tract, oral peptide drugs encounter substantial physiological barriers leading to low oral bioavailability. Therefore, peptide drugs are mostly administered by parenteral routes. Although parenteral delivery of peptide drugs achieves high bioavailability, this is associated with inconvenience and discomfort, even causing severe side effects compared with the oral route possessing a high degree of patient compliance. Therefore, numerous studies concentrate on novel strategies to improve the oral bioavailability of peptide drugs. Some delivery technologies such as Eligen™ and Axcess™ have been successfully applied to the oral dosage form of therapeutic peptides and have accelerated relevant oral formulations for Food and Drug Administration (FDA) approval and clinical treatment. In this review, we focus on the oral peptide delivery, mainly summarizing the progress of recent strategies used to overcome oral barriers and the commercialization applications of related patents, which could facilitate the research and development (R&D) of clinical applications of oral delivery techniques for peptide drugs.

OBJECTIVEThe purpose of the present study was to explore the relationship between the expression of S100A4 and E-cad protein and some clinicopathological factors such as histological types, TNM stages, lymph node metastasis and prognosis in non-small cell lung cancer (NSCLC), and analyze whether there is a correlation between them.

METHODSThe expression of S100A4 protein and E-cad protein was detected with immunohistochemical SP technique in 116 non-small-cell lung cancers.

RESULTSThe positive immunohistochemical staining for S100A4 protein was observed in 64 out of the 116 patients, with a positive rate of 55.2%. The expression rate of S100A4 protein was associated with age, tumor size, lymph node metastasis and prognosis of NSCLC. The expression of S100A4 protein was not significantly related with histological types, sex and TNM stages. The positive rate of E-cad protein expression was 65.5%. The expression of E-cad protein was associated with TNM staging, lymph node metastasis and prognosis of NSCLC. The expression of E-cad protein had no significant correlation with histological types, patient age and sex. An inverse correlation between the levels of S100A4 and E-cad protein expression was found (P < 0.005).

CONCLUSIONExpression of S100A4 protein and loss of E-cad protein expression are significantly associated with tumor progression, and may become valuable markers in prediction of biological behavior and tendency of metastasis of non-small-cell lung cancers.

Biomarkers, Tumor ; Cadherins ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; S100 Calcium-Binding Protein A4 ; S100 Proteins ; metabolism ; Survival Rate

Objective To study the animal toxicity test on skin of the Chinese herbal drugs liquid paste (CHDLP).Methods Acute and multiple times skin stimulation test and skin allergy test were performed on healthy guinea pig by using CHDLP.Results The acute skin stimulation score and intensity score were all zero.Multiple skin stimulation score and pathological examination score were all zero.So were the skin allergy score and the sensiti- zation rate.Conclusion CHDLP is a weak sensitizer and arose no skin stimulation on guinea pig.It's safe when ex- ternal use.

Esophagogastric Junction/surgery* ; Gastrectomy/methods* ; Humans ; Laparoscopy/methods* ; Retrospective Studies ; Stomach Neoplasms/surgery*

AIM: To observe the effect of endothelin-1(ET-1) on the cytoskeleton protein F-actin of cultured human trabecular meshwork (HTM) cells. METHODS: Cultured HTM cells were randomly divided into four groups: control group(0mol/L), low-dose ET-1(10-9mol/L) treatment group, middle-dose ET-1(10-8 mol/L) treatment group, and high-dose ET-1(10-7 mol/L) treatment group. After treated with ET-1, the expression of cytoskeleton protein F-actin in trabecular meshwork was analyzed with Western-blot and the distribution of F-actin was detected with FITC-Phalloidin probe. RESULTS: ET-1 dose-dependently and significantly increased F-actin in trabecular meshwork cells. The F-actin stress fiber and periphery actin fiber highly increased and manifested mild reorganization after treated with ET-1; and there were much more cell-to-cell and cell-to-extracellular matrix attachments formation in ET-1 treated HTM cells than that in the untreated HTM cells. CONCLUSION: ET-1 promoted the expression of cytoskeleton protein F-actin and induced the trabecular meshwork actin cytoskeleton reorganization.

Objective To evaluate the protective effect of Yersinia pestis capsular antigen F1 and recombinant rV270 on mice after immunization with them.Methods According to body weight,40 female Balb/c mice aged 6 to 8 weeks were randomly divided into four experimental groups(Fl-10 μg + aluminum adjuvant,F1-20 μg + aluminum adjuvant,rV-10 μg + aluminum adjuvant,and rV-20 μg + aluminum adjuvant) and a control group,8 in each group.Mice in experimental groups were immunized with the natural antigen F1 and recombinant antigen rV270 adsorbed to 25％ aluminum adjuvant and the control group was immunized with the same amount of aluminum adjuvant.Each mouse was immunized at the hind leg muscle with 100 ml immunizing agent,then a booster immunization was done once on the 21st day after the first immunization.The blood of all mice was collected on the 8th week after the first immunization,serum antibody titers were detected by ELISA and the data of antibody titers were analyzed by t test for comparison between groups.At the same time the mice were injected subcutaneously with 2000-fold LD50 of Yersinia pestis virulent strain 141,after 14 days,the protective effect of immunization was analyzed.Results The control group did not produce antibody.Antibody geometric mean titers (GMT) of the F1-10 mg + aluminum adjuvant and F1-20 mg + aluminum adjuvant groups were 1 ∶ 30443.9,and 1 ∶21527.8,respectively,and compared between the two groups,the difference was not statistically significant (t =1.1282,P ＞ 0.05).The GMTs of the rV-10 μg + aluminum adjuvant and rV-20 μg + aluminum adjuvant groups were 1 ∶ 13957.3 and 1 ∶18100.9,respectively,and compared between the two groups,the difference was not statistically significant(t =0.9408,P ＞ 0.05 ).After subcutaneous injection with Yersinia pestis virulent strain 141,all mice died in the control group but all survived in the experimental group.Conclusion The immune activity of natural antigen F1 and recombinant rV270 is high,which can be used as the main component of subunit vaccine in the plague subunit vaccine study.

Objective To understand the effect of hyaluronic acid (HA) on the proliferation and apoptosis of chondrocytes cultured in vitro with Kashin-Beck disease(KBD) to provide the experimental evidences for treating KBD diseases with HA. Methods The articular cartilage samples collected from KBD patients were selected according to Diagnosis for Kaschin-Beck Disease(GB 16003-1995). And the normal cartilage samples were collected from victims of incidence (control). Chandrocytes were separated and cultured in vitro. Then varying dosages of HA were administered to chondrocytes and individed into 0,100,500 mg/L group, according to HA doages. The effect of HA on the proliferation and apoptosis of chondrocytes cultured/n vitro both KBD and the controls were investigated by methyl thiazolyl tetrazolium(MTT), Annexin V/PI staining on 2~(nd), 4~(th), 6~(th) day. Results In the control group, 500 mg/L group(0.140 ± 0.049) promoted chondrocyte proliferation significantly than 0 mg/L group (0.116 ± 0.021 ) at the 4~(th) day(P < 0.05), similar phenomenon was observed in KBD group in the 6~(th) day between 500 and 0 mg/L group(0.179 ± 0.081,0.128 ± 0.017, P< 0.05). In the KBD group, compared with 0 mg/L (12.860 ± 2.159), both 100 and 500 mg/L( 10.458 ± 1.143,7.877 ± 1.346) inhibited chondrocyte apoptosis rate (P < 0.05). In control, apoptosis rate of 500 mg/L group(4.045 ± 1.204) descreased compared with 0 mg/L group (7.128 ± 1.244, P < 0.05). Conclusion HA can promote the proliferation and inhibit the apoptosis of KBD chondrocytes cultured in vitro, and 500 mg/L HA play more effective role than that of 100 mg/L in promoting proliferation and inhibiting poptosis.

Abstract: Objective　To investigate the clustered regularly interspaced short palindromic repeats (CRISPR) genotypes and regional distribution of Yersinia pestis strains in the natural plague foci of Hainan Tibetan Autonomous Prefecture of Qinghai Province (referred to as "Hainan prefecture") and provide a scientific basis for plague prevention and control in this area. Methods　A total of 36 representative Yersinia pestis strains, which were isolated from different host animals and insect vectors from 1954 to 2009 in Hainan Prefecture, were selected as experimental subjects. The DNAs were extracted using the traditional sodium dodecyl sulfate decomposition and phenol-chloroform method. Three pairs of CRISPR primers (YPa, Ypb, YPc) were used for PCR amplification, sequencing and analysis of the DNA of the tested strains, respectively, as a means to identify the CRISPR genotypes of Yersinia pestis in Hainan Prefecture. Results　A total of 17 spacers were observed among 36 strains of Yersinia pestis, including 9 of YPa, 5 of YPb and 3 of YPc. All strains were divided into 5 CRISPR gene clusters (Cb2, Cb4 ', Ca7, Ca7 ', Ca35 ') and 6 genotypes (G1, G9, G22, G22-A1 ', G26-A1 ', G26-A1 'A4 -). The G26-a1 ' was the main genotype, which was distributed in Gonghe, Guide and Xinghai County, and the G22 is the second type, which was distributed in Gonghe and Guide County. Conclusions　The genetic polymorphism of CRISPR loci of Yersinia pestis strains in Hainan was high, and the regional distribution characteristics of Yersinia pestis strains with different genotypes were significant.

Abstract: Objective To investigate the current status of streptomycin resistance of Yersinia pestis caused by point mutations of rpsL gene in Qinghai, so as to provide theoretical basis for precise clinical medication and prevention of drug resistance of human plague outbreak in South area of Qinghai Province in the future. Methods A total of 104 representative strains of Yersinia pestis collected from plague patients, vector insects and intermediate hosts in South area of Qinghai Province from 1957 to 2009 were screened, isolated and cultured by Hiss agar plates. The DNA of representative Yersinia pestis was extracted by sodium dodecyl sulfate lysis and phenol-chloroform method. The primers forward primer and reverse primer and TaqMan-MGB probes probe1 [FAM] and probe2 [VIC] were designed for the rpsL gene of streptomycin resistance gene in China. Real-time PCR with TaqMan-MGB fluorescent probe was used to detect the mutations of rpsL gene in streptomycin resistance locus of 104 strains of Yersinia pestis in South area of Qinghai Province. Results The FAM test results of 104 strains in South area of Qinghai Province were positive, corresponding to the detection of rpsL (128 : A ), RFU peak >1 000,negative <200. VIC test results of all tested strains were negative, corresponding to the detection of rpsL (128:G), RFU peak <200, positive >1 000. That is, no strains with rpsL gene mutation related to streptomycin resistance were found in the 104 strains of Yersinia pestis in Qingnan Province. Conclusion This study provides basic data on the distribution of streptomycin resistance of Yersinia pestis in South area of Qinghai Province, and lays a foundation for preventing the occurrence of drug resistance and clinical treatment of Yersinia pestis in South area of Qinghai Province.