The plasmid pXJ-41-p15, which contains the full length DNA coding for p15 was introduced into human melanoma cell line A375 in which p 15 was deleted by DNA recombination and transfection. Using G418, the positive clones were selected. And the cell model overexpressing p15 was constructed suc cessfully through the analysis of PCR and Western blot. It is showed tha t the expression of p15 was further enhanced after the cells overexpressing p15 were treated with PMA for 72 hours. In contrast of the control cells, the PKC a ctivity was further declined in the cells overexpressing p15 after treated with PMA. At the same time, the growth rate of cell was decreased more significantly and approximate 30% apoptotic cells were found. The expression of Caspase-3(P2 0) was increased in the apoptotic cells. It is indicated that CKI p15 was relat ed to PKC signal transduction in the regulation of cell proliferation and apopto sis. They may be involved in the apoptotic pathway including Caspase3, thus indu cing the apoptosis of cells.

The intracellular signal transduction pathways play an important role in the occurrence and development of thyroid cancer.Mitogen activated protein kinase (MAPK) signal transduction pathway,phosphatidylinositol 3-kinase (PI3K)-AKT pathway,NF-κB pathway and RASSF1-MST1-FOXO3 pathway affect the occurrence and development of thyroid cancer by modulating the activity and expression of specifical proteins,and determine the growth and apoptosis of thyroid cancer cells.

The synchronized HeLa cells were used to study the effect of protein kinase A(PKA) inhibitor on the progression of S phase. Synchronized cell s in S phase were obtained by the method of TdR double block through 3H-T dR incorporation assay. The PKA inhibitor typeⅢ obviously increased the level of 3H-TdR incorporation of S phase in HeLa cells. In contrast with contro l, the activity of thymidine kinase (TK) in S phase increased, too. It indicated that PKA played an inhibitory role in S phase progression of HeLa cells. With t he method of Western blotting, the PKA inhibitor typeⅢ enhanced the level of C yclinA and PCNA, inhibited the expression of p21, which is a negative regulator of cell cycle, but had no effect on the expression of CDK2. The results showed t hat PKA could negatively regulate the S phase progression by affecting the level of CyclinA, PCNA and influencing the expression of p21 protein. This may be one of the molecular mechanisms which is involved in the negative regulation of S p hase progression by PKA in HeLa cells.

Objective To study the effect of mechanical tension on collagen arrangement and illustrate the relationship between tissue architecture and tension properties. Methods Cell morphologies, orientation and collagen arrangement of fibroblasts cultured in three different types of collagen gels with variation of mechanical tension were observed by phase contrast photomicrographs, light microscopy and transmission electron microscopy. Expression and distribution of a-smooth muscle actin (α-SMA) were investigated by immunohistochemistry. Results Phase contrast photomicrographs, light microscopy and transmission electron microscopy showed high level of tension distributed anisotropically in the monolayer gels and the anchored collagen gels, with bipolar shape of the fibroblasts, obvious polarity, arrangement of exogenous collagen fibres parallel to the long axis of the fibroblasts, especially prominent in monolayer gels. Low level of tension distributed isotropically was observed in floating collagen gels, with stellate morphology and arrangement of exogenous collagen fibres in a reticular array. Immunofluorescence showed that fibroblasts expressed high level of α-SMA protein distributed along the long axis of fibroblasts in the monolayer gels and the anchored collagen gels, especially in former ones. In contrast, few expression of α-SMA protein was found in floating collagen gels. Cell morphologies and orientation, expression and distribution of α-SMA as well as collagen arrangement of fibroblasts in the monolayer gels and the anchored collagen gels were similar to those in granulation tissue, whereas floating collagen gels resembled normal dermis or remodelled tissues. Conclusions Tissue architecture or morphology of the dermis are corresponding to tension proporties. Different tissue architectures are closely correlated with particular tension proporties.

This study was aimed to search and analyze patents of Chinese herb published both at home and abroad in 2010. The Derwent World Patents Index (DWPI) was used as database source. A total of 8 704 Chinese herb patent records were retrieved. And the geographical distribution was concentrated in China. The technological field was mainly on hepatitis, liver cirrhosis, lower back pain, cough and other diseases. Innovations on formulation and preparation method also required special attention. In addition, through the analysis on patent attribution agency, some well-known Chinese enterprises, who aim to gain the initiative in specific technical areas, have actively applied a number of patents in specific areas.

Objective To detect the histological characteristics of collagen nodules from hypertrophic scars (HS) and investigate the origin of collagen nodules.Methods The scar tissues were collected from patients with plastic operation.Morphological characteristics of collagen nodules were observed by light microscopy of HE-stained sections; expressions of type Ⅰ /Ⅲ collagens were observed by polarized light microscopy of sirius red-stained sections; expression and distribution of myofibroblasts (MFb)-specific protein (α-smooth muscle actin,α-SMA) were observed by immunostaining in order to observe level of local tissue tension.Results Collagen nodules varied in shape,not only sphereshaped,and in size.Moreover,abundant fibroblasts (Fb) with large and light-stained nuclei were seen in the nodules compared to non-nodule area,indicating that the cells located in the modules were active.Some collagen nodules were composed largely of collagen type Ⅲ (green),but some mainly contained collagen type Ⅰ (red or yellow),indicating the difference in the time of nodule formation.α-SMA was expressed mainly in the deep dermis equivalent to the level of reticular layer of the new scar tissues (2 months after injury) ; α-SMA was expressed mainly in the nodules of the old scar tissues (2-10 years after injury),but almost not in non-nodule areas except for a strongly positive staining in the vessels.Moreover,α-SMA presented a heterogeneous distribution in the collagen nodules,with stronger expression in the epidermal end than in the subcutaneous tissue end and stronger expression in the superficial dermis than that in the deeper part.It was suggested that there existed massive amount of MFb and high tension in the nodules arid that the tension distribution was not uniform in or between the nodules.Conclusions Collagen nodules are of varying shape and size and collagen types are associated with the time of nodule formation.Moreover,Formation of the collagen nodules may be closely related to the distribution and evolution of the local tissue tension.

Objective:The present study was aimed to determine the value of cone beam CT (CBCT) in predicting the risk of lingual bone plate injury during extraction of impacted mandible third molar (IMTM).Methods:The original CBCT data of 150 teeth (50 in vertical, 50 in angular and 50 in horizontal ) in January 2018 to December 2019 in Panzhihua Central Hospital of Sichuan Province were collected and analyzed. The thickness of lingual bone plate in enamel cementum boundary (ECB), root middle (RM) and root tip (RT) of each IMTM was measured by the software of CBCT system, and datas were analyzed by one-way ANOVA.Results:The average thickness of lingual bone plate in ECB of IMTM was (1.36±0.43)mm, (1.21±0.44)mm and (1.28±0.40)mm in vertical, horizontal and angular groups, respectively, with no significant difference ( F=1.07, P=0.35). The average thickness of lingual bone plate in RM of IMTM was (1.48±0.33)mm, (1.06±0.57)mm and (1.11±0.45)mm, respectively, with statistically significant difference ( F=8.78, P<0.01). The average thickness of lingual bone plate in RT of IMTM was (1.44±0.49)mm, (0.84±0.58)mm and (0.86±0.64)mm, respectively, with statistically significant difference ( F=12.35, P<0.01). Compared with the mandibular second molar, there were statistically significant differences in the average thickness of the lingual bone plate in ECB ( F=5.03, P<0.01), the RM ( F=15.13, P<0.01) and the RT ( F=33.12, P<0.01) of the IMTM among the three groups. In addition, the horizontal and angular IMTM, the thinness of lingual bone plate in RT region was more likely to occur than in vertical, and the absence of lingual bone plate was most likely to occur in patients with partial buccal crown. Conclusions:The doctor-patient communication and risk prediction should be sufficient before IMTM extraction when CBCT shows that the lingual bone plate of RT region is thin or absent. At the same time, we should avoid violent operation and thoroughly protect the lingual bone plate in the process of tooth extraction, and guard against serious complications such as perforation or fracture of lingual bone plate of mandible, and root displacement.

Recently, herbal medicine including traditional Chinese medicine (TCM) has gained huge attention in the world. In 2015, the global trades of herbal medicine reached 93.15 billion US dollars. And, the latest statistics from the Ministry of Industry and Information Technology of People's Republic of China showed that total sales of Chinese patent medicine and raw herbs reached 120 billion US dollars in 2014. Therefore, the aim of this study was to analyze the situation of international marketing on herbal medicine and how much TCM shared in it. The PubMed database, search engines and government websites and research reports were searched for analyses. The results showed that total trades of TCM products in both domestic and foreign markets, were about 135 billion US dollars, including Chinese patent medicine, raw herbs, herbal extracts, herbal health care products, whose proportion of the global marketing was 80%.

Objective To simulate the environmental factors during the process of formation and evolution of hypertrophic scar,so as to explore the effects of moderate and severe hypoxia and low concentration of serum protein on the function of human hypertrophic scar fibroblasts (HSFs).Methods Human HSFs were routinely cultured.Cells of the 3rd to the 6th passage were divided into 10.0％ oxygen + 10.0％fetal calf serum (FCS),5.0％ oxygen + 5.0％ FCS,and 0.5％ oxygen + 0.5％ FCS groups according to the random number table.After being cultured with DMEM nutrient solution with no FCS for 24 h,the cells were cultured with the corresponding volume fraction of oxygen and FCS.Cell proliferation activity was determined with methyl-thiazole-tetrazolium assay (denoted as actual cell number).Content of total collagen was detected with Sirius red staining method (denoted as absorbance value).Protein expression levels of hypoxia-inducible factor 1 α (HIF-1 α),vascular endothelial growth factor (VEGF),transforming growth factor β1 (TGF-β1),B-cell lymphoma 2 (Bcl-2),and P53 were determined with Western blotting (denoted as ratio of gray value).Cell apoptosis rate was detected by in situ end labeling method.The sample numbers of each group in the above experiments were all 3.Data were processed with Kruskal-Wallis test and Dunnett test.Results (1) Compared with 11 000 ± 1 306 in 10.0％ oxygen + 10.0％ FCS group,the cell proliferation activity was higher in 5.0％ oxygen + 5.0％ FCS group (13 290 ± 1 500,P ＜ 0.05),but lower in 0.5％oxygen +0.5％ FCS group (6 999 ±765,P ＜0.05).(2) Compared with 0.039 6±0.004 2 in 10.0％oxygen + 10.0％ FCS group,the content of total collagen of cells was higher in 5.0％ oxygen + 5.0％ FCS group (0.051 6±0.005 1,P ＜0.05),but lower in 0.5％ oxygen +0.5％ FCS group (0.015 6 ±0.002 4,P ＜ 0.05).(3) Compared with those in 10.0％ oxygen + 10.0％ FCS group,the protein expression levels of HIF-1 α,VEGF,TGF-β1,and Bcl-2 were increased (with P values below 0.05),with no obvious difference in protein expression level of P53 in 5.0％ oxygen + 5.0％ FCS group (P ＞ 0.05),whereas the protein expression levels of HIF-1α,VEGF,TGF-β1,and Bcl-2 were decreased (with P values below 0.05),while the protein expression level of P53 was increased in 0.5％ oxygen + 0.5％ FCS group (P ＜0.05).(4) Compared with (1.2 ±0.9)％ in 10.0％ oxygen+ 10.0％ FCS group,the cell apoptosis rate in 5.0％ oxygen + 5.0％ FCS group showed no significant difference [(2.6 ± 0.9)％,P ＞ 0.05],while it was significantly increased in 0.5％ oxygen + 0.5％ FCS group [(13.3 ± 4.1)％,P ＜ 0.05].Conclusions Severe hypoxia and low concentration of serum protein can inhibit proliferation activity and production of total collagen of human HSFs and induce their apoptosis.

Objective To evaluate the clinical implication of circulating endothelial microparticles in severe burned patients in burn shock resuscitation period and in patients with severe infection.Methods A retrospective case control study was conducted on 90 burned patients from January 2009 to December 2015.The patients were divided into burn shock group (n =57) and infection group (n =33).The infection group was further divided into sepsis group (n =18) and non sepsis group (n=15).There were 17 healthy controls.The numbers of CD31 +,CD51 +,CD144 +,CD54 +,and CD62E + EMPs in plasma were measured by flow cytometry immediately thereafter.The numbers and ratios of EMPs in plasma of the 16 patients from burn shock group were compared before and after fluid resuscitation.The numbers and ratios of EMPs in plasma were compared among groups.Sequential Organ Failure Assessment (SOFA) scores were used to assess the burn severity in patients at enrollment.The numbers and ratios of EMPs in plasma were compared between sepsis and non sepsis group.The correlations of SOFA with the numbers of EMPs and with the burn size were analyzed.Results Compared with control group,the numbers of CD31 +,CD51 +,CD144 +,CD54 +,and CD62E + EMPs were increased significantly in both burn shock group and infection group (P ＜0.05 or ＜0.01);the ratios of CD31 + EMPs,CD51 + EMPs,and CD54 + EMPs in plasma were significantly higher in burn shock group (P＜O.05 or ＜0.01);the ratios of CD31 + EMPs and CD54 + EMPs were significantly higher in infection group (P ＜ 0.05 or ＜ 0.01).The number of CD62E + EMPs,the ratios of CD144 + EMPs and CD62E + EMPs were higher in burn shock group than those in infection group (P ＜0.05 or ＜0.01).The ratio of CD31 + EMPs in plasma was significantly higher before resuscitation than that after resuscitation in burn shock period patients (8.8 ± 1.7 vs.5.2 ±0.9) (P ＜0.05).The ratios of CD31 + EMPs and CD54 + EMPs were significantly higher in sepsis group than those in non-sepsis group (8.0± 1.3 vs.4.5 ± 1.0;18.3 ± 2.2 vs.8.9 ± 1.4) (P ＜ 0.05 or ＜ 0.O1).The number of antigen-positive EMPs correlated significantly with SOFA score (P ＜0.05 or ＜0.01).The ratio of CD54 + EMPs correlated significantly with SOFA score (P ＜0.01).The number and the ratio of CD54 + EMPs as well as the ratio of CD144 + EMPs was correlated with bum size (P ＜ 0.05).SOFA score was positively correlated with the burn size (P ＜ 0.01).Conclusions Expressions of CD31,CD54,CD144,CD62E antigens EMPs might play a role in burn pathological damage and serve as an indicator of endothelium function change after bums.Expression of CD54 of EMPs plays a role in assessing the function and severity of burned organs.