Objective To explore the possible pathway and regulatory mechanism of dermal fibroblasts' transdifferentiation into myofibroblasts induced by estrogen. Methods Human dermal fibroblasts were divided into six groups (A: control; B: estrogen; C: estrogen + ICI-182780; D: estrogen + SB203580; E: estrogen + PD98059; F: estrogen + SP600125). The cells were collected for RNA extraction and the expression of α-SMA was detected by real time quantitative RT-PCR. Some cells were analyzed by single cell RT-PCR to detect positive expression percentage of α-SMA.Results The expression and positive rate of α-SMA in estrogen group were significantly increased (Group B vs. Group A, 7. 385±0. 246 vs 1. 367±0. 034, P＜0.01) and those in ICI-182780 group and SP600125 group were significantly inhibited (Groups C and F vs. Group B, 4. 619 ±0. 164,2. 409±0. 091 vs 7. 385±0. 246, P＜0. 05). Conclusions In the process of fibroblast transdifferentiation into myofibroblasts induced by estrogen, estrogen β receptor and JNK-MAPK signal transduction pathway may play an important role.

From the standpoint of training mode,the paper makes an effort to analyze the present situation on Sino-Foreign cooperation in nnning schools in mode of nursing personnel training in Hubei Province,find out the existing problems and put forward the corresponding proposals in order to ensure sustainable development of the Sino-Foreign cooperation in running schools in mode of nursing personnel training.

A sensitive, rapid and specific liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method for quantification of gabapentin in human plasma has been developed. After a single plasma protein precipitation with methanol, gabapentin and metformin (internal standard) were chromatographed on a Inertsil ODS-3 column (50 mm x 2.1 mm ID, 3 microm) with mobile phase consisting of methanol-0.2% formic acid aqueous solution (80:20, v/v) at a flow-rate of 0.2 mL x min(-1). Electrospray ionization (ESI) source was applied and operated in the positive ion mode. Multiple reaction monitoring (MRM) mode with the transitions of m/z 172 --> m/z 154 and m/z 130 --> m/z 71 were used to quantify gabapentin and metformin, respectively. The run time was 2.2 min. The linear calibration curve was obtained in the concentration range of 40.8-8.16x10(3) ng x mL(-1). The lower limit of quantification was 40.8 ng x mL(-1). The intra- and inter-day precision (RSD) was less than 12%, and the accuracy (RE) was within +/-6.4% calculated from quality control (QC) samples. The method was used to determine the concentration of gabapentin in human plasma after a single oral administration of 600 mg gabapentin capsule to 20 healthy male Chinese volunteers. The method was proved to be selective, sensitive, rapid and suitable for pharmacokinetic study of gabapentin in human plasma.

OBJECTIVE:To explore the cultivation method for community pharmaceutical care talents in higher vocational colleg-es. METHODS:According to the talent training aims to meet the demand of community pharmaceutical service positions in higher vo-cational colleges,the teaching reform was carried out for talent cultivation mode,course structure system,training base construction and teaching methods. RESULTS:In terms of talent cultivation mode,the work-integrated learning mode was built;in terms of course structure system,“Medicine,Pharmacy,Humanities and Expansion”course module was constructed to further optimize it;in terms of training base construction,the capital investment was increased to improve the school training base,meanwhile,cooperation between schools and enterprises was carried out to strengthen the construction of out-of-school training bases;in terms of teaching methods, PBL,case teaching model and situational teaching method were implemented. CONCLUSIONS:The quality of pharmaceutical talents has been significantly improved by implementing reform of the talent cultivation mode,the course structure system and teaching meth-ods,together with constructing training bases.

Adolescent ; Adult ; Aged ; Endoscopy ; Female ; Humans ; Male ; Middle Aged ; Nasal Septum ; Nose Neoplasms ; surgery ; Young Adult

Objective To explore the feasibility of using irreversible electroporation (IRE)mediating HPV16 E6 shRNA into cervical cancer cell line SiHa,and to clarify the influence of their co-effect on the proliferation of SiHa cells and its mechanism.Methods A HPV16 E6 gene specific interference sequence was inserted in pGenesil-1 to build a interference vector.10 pulses of IRE with 800 V,100 μs,and 1 Hz were applied to the suspension of SiHa cells and vectors.According to the treatment factors,control group,IRE group,pGenesil-N group,pGenesil-N+IRE group,pGenesil-E6 group and pGenesil-E6 + IRE group were set up.The expression of green fluorescent protein (GFP)and transfection efficiency were confirmed by inverted fluorescence microscope 24 h after the vector was transfected by IRE,and the expression efficancy of GFP was calculated.The expression levels of E6 mRNA and protein were detected by RT-PCR and Western blotting method which was also applied to detect the expressions of P53 and PCNA.The proliferative activity of SiHa cells was determined by CCK-8 assay.Results Enzyme digestion and DNA sequencing verified that the vectors were correctly constructed.GFP was seen under inverted fluorescence microscope 24 h after IRE transfection.Compared with IRE group,the expression levels of E6 mRNA and protein were decreased detected by RT-PCR and Western blotting method after the vectors were treated with IRE,the P53 protein expression level was increased (P < 0.05),and the PCNA expression level was decreased (P <0.05).The CCK-8 assay results showed the proliferative activity of SiHa cells in pGenesil-E6+IRE group was decreased more obviously than that in pGenesil E6 group (P <0.05).Conclusion IRE can play the role of gene transfection of mediating HPV16 E6 shRNA into SiHa cells, and their co-effect can significantly inhibit the proliferation of SiHa cells.

Objective In order to analyze the variation of HA genes of influenza viruses (H1N1) by being compared with the vaccine strain A/California/07/2009(H1N1) recommended by WHO ,influenza viruses (H1N1) isolated from 2009 to 2014 were selected to do this study .Methods According to the different isolating time and place ,47 strains of H1N1 collected from 2011 to 2014 were selected .Then the 47 strains′ nucleotide sequence of HA genes which were sequenced in the study and other 25 se‐quences of HA genes which were sequenced in 2009 were collected .Nucleotide and amino acid sequences were analyzed by using molecular biology software ,and the phylogenetic trees were drawn .Results A total of 72 strains isolated from 2009 to 2014 were closely related to the vaccine strain A/California/07/2009(H1N1) ,the nucleotide variance and amino acid variance between the 72 strains were 0-2 .7% and 0-3 .1% respectively .Compared with the vaccine strain A/California/07/2009(H1N1) ,the nucleotide variance and amino acid sequence variance were 0 .4% -2 .4% and 0 .9% -3 .1% respectively .The amino acids sequence indicated that ,although the variance was increased by years ,the H1N1 viruses were still showed characteristics of low pathogenic influenza viruses .It was also found that there were 9 strains lost their potential glycosylation site at HA protein site 481 in 2009 ,while in 2013 there were 6 strains got new potential glycosylation sites at HA protein site 162 .Conclusion The vaccines (H1N1) recom‐mended by WHO was still protective to people in Chongqing .But as time goes by ,antigen drift may occur in some new antigenic drift strains and the routine monitoring of influenza viruses should be continued .

Objective To investigate the effect of expanded deltopectoral perforator flap as a free skin flap for distant transfer to aesthetically repair large facial scars via microsurgical technique.Methods 20 cases of large facial scars were treated with free expanded deltopectoral skin flaps via microsurgical technique.At the first stage,24 large volume expanders were inserted into the subcutaneous pockets in the deltopectoral zone.4 to 6 months later,the second stage operation was performed when the flaps were expanded fully and facial scars removed;deltopectoral perforator flaps harvested choosing the second or the third thoracic artery perforating branches as the pedicle;the surface of the wound was covered with the flaps.Results All of the 24 flaps survived completely and the wounds healed well after transplantation.The 6 months to 2 years' follow-up results showed that the color and texture of the flaps matched the recipient place well without swelling.The superficial and deep sensation recovered very fast.Conclusions It is safe,practical and effective that the delto-pectoral perforator flaps are used to repair large facial scars avoiding the shortcomings of traditional pedicle transfer such as more procedures,longer fixed time,or more wasted expanded skin.Free transfer of expanded delto-pectoral perforator flaps can achieve ideal function and aesthetic effects.

Objective To explore the effects of silencing Livin gene by RNA interference mediated by lentiviral vector on colorectal cancer HT-29 cell xenograft growth and sensitivity to radiotherapy in nude mice.Methods BALB/c nude mice models were established by subcutaneously inoculating differently treated HT-29 cells into nude mice and the tumor growth situation of tumors was observed by measuring the volume of tumors and the weight of the nude mice at different time points after cell seeding.Livin expression was detected by RT-PCR and immunohistochemistry,respetively.Apoptosis rate was detected by TUNEL.Normal saline,lentivirus carring unrelated sequences,lentivirus caning Livin shRNA were injected intratumorally.All the nude mice were given 10 Gy of 6 MV X-ray irradiation.The changes of mice weight and the tumor volume were measured at different time points and the weight and tumor growth curves were drawn.Results The inhibition rate of tumor volume was(50.04 ± 0.07)％ and the tumor weight of the RNA interfering group was significantly less than that in experimental group compared to the blank and negative groups(F=4.85,P＜0.05),and the inhibition rate of tumor weight was(50.27 ±0.17)％.Relative Livin mRNA expression level in the RNA interfering experimental group was(17.75 ±0.08)％,and was significantly lower than that of the blank group(67.60 ± 0.05)％ and the negative group(68.54 ± 0.03)％(F=89.97,P＜0.01).Livin protein expression level in the RNA inferring group was also significantly lower[(36.00 ± 3.40)％ versus(85.00 ± 3.15)％,(80.33 ± 3.08)％,F=107.32,P＜0.01].The apoptosis rate in the RNA interfering experimental group was significantly higher than that in the blank and the negative groups[(23.67 ± 2.25)％ versus(5.00 ± 1.50)％,(8.33 ± 1.82)％,F=56.94,P＜0.01].Combined with radiotherapy,the tumor volume at different groups had significant difference(F=10.70,P＜0.01),and RNA interfering group was significantly less than negative group and blank group(F=7.01-9.32,P＜0.01).Conclusions Silencing of Livin gene expression by lentiviral vector-mediated RNA interference could inhibit the growth of colorectal HT-29 cell xenograft and increase the sensitivity of the transplanted tumors to radiotherapy.

Purpose To investigate the relationship and clinical significance of high-risk human papillomavirus infection and expression of c-jun and c-fos protein in cervical carcinoma.Methods Cervista HR-HPV testing,immunohistochemical SP method were employed to detect the infection of HR-HPV,c-jun and c-fos protein in 70 cases of CSCC,60 cases of cervical intraepithelial neoplasia and 20 cases of chronic cervicitis.The correlation between c-jun and c-fos protein expression and of HR-HPV infection was analyzed.Results Of the 70 cases of CSCC,the positive HR-HPV was 69 cases,the positive rate in HR-HPV in A9 group was the highest (85％,59/69).The positive expression of c-jun and c-fos were 80％ (56/70),85.7％ (60/70) in 70 cases CSCC,70％ (21/30) and 70％ (21/30) in 30 cases of cervical high grade intraepithelial neoplasia,and 20％ (6/30),23.3％ (7/30) in 30 cases cervical low grade intraepithelial neoplasia,but the positive expression rates of c-jun and c-fos in the 20 cases of chronic cervicitis were 0.Expression of c-jun and c-fos in cervical cancer group was higher than that in the chronic cervicitis group and low grade intraepithelial neoplasia (P ＜ 0.05).The expression of c-jun and c-fos was statistically significant in different groups of clinical stage and pathologic grading in the CSCC (P ＜ 0.05).Additionally,the expression of c-jun and c-fos was positively correlated with the infection of HR-HPV in CSCC (P ＜ 0.01).Conclusion The infection of HR-HPV has significant subtype-specific and has a positive correlation with the expression of c-jun and c-fos in CSCC,which suggests that AP-1 pathway activation after HRHPV infection may be associated with the occurrence and development of cervical carcinoma.