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 investigate the effects of denatured collagen type Ⅰ on differentiation of human fibroblasts into myofibroblasts. Methods: A small amount of normal skin donated by burn patients undergoing scar surgery was collected. Human fibroblasts were obtained by method of explant culture and then sub-cultured. The fourth passage of cells were used in the following experiments. (1) Fibroblasts were divided into normal collagen group and denatured collagen group according to the random number table, with 10 wells in each group. Fibroblasts in normal collagen group were cultured on normal collagen type Ⅰ coated coverslips. Fibroblasts in denatured collagen group were cultured on denatured type Ⅰ collagen coated coverslips. Expression of proliferating cell nuclear antigen (PCNA) was detected by immunohistochemical method, and the percentage of PCNA positive cells was calculated. (2) Another batch of fibroblasts were grouped and treated as in (1), with 12 wells in each group. Proliferation activity of cells was determined with methyl-thiazolyl-tetrazolium colorimetry method. (3) Another batch of fibroblasts were grouped and treated as in (1), and the microfilament morphology of cells was observed by rhodamine-phalloidin staining. (4) Another batch of fibroblasts were grouped and treated as in (1). Expression of α smooth muscle actin (α-SMA) of cells was detected by immunohistochemical method, and expression of OB-cadherin of cells was detected by immunofluorescence method. (5) Another batch of fibroblasts were divided into normal collagen, denatured collagen, and common coverslips groups according to the random number table, with 6 wells in each group. Fibroblasts in normal collagen and denatured collagen groups were treated as in (1), while fibroblasts in common coverslips group were cultured on coverslips without collagen coating. Expressions of α-SMA and OB-cadherin of cells were determined with Western blotting. (6) Another batch of fibroblasts were grouped and treated as in (5), and then the mRNA expressions of collagen type Ⅰ, collagen type Ⅲ, and α-SMA of cells were determined with real-time fluorescent quantitative reverse transcription polymerase chain reaction. Data were processed with t test, one way analysis of variance, and least-significant difference test. Results: (1) The percentage of PCNA positive cells in denatured collagen group was (83±9)%, significantly higher than (29±9)% in normal collagen group (t=13.53, P<0.01). (2) The proliferation activity of fibroblasts in denatured collagen group was 0.32±0.06, significantly higher than 0.25±0.05 in normal collagen group (t=3.06, P<0.01). (3) The microfilament of fibroblasts in normal collagen group was arranged vertically and in parallel way, paralleling the long axis of cells. The microfilament of fibroblasts in denatured collagen group was denser and thicker. (4) Most fibroblasts in normal collagen group showed long shuttle-like shape typically. Morphology of fibroblasts in denatured collagen group changed, and cells were obviously spreading. Expressions of α-SMA and OB-cadherin of fibroblasts in denatured collagen group were stronger than those in normal collagen group. (5) Expressions of α-SMA of fibroblasts in denatured collagen, normal collagen, and common coverslips groups were respectively 1.69±0.41, 0.89±0.27, and 1.46±0.42. Expression of α-SMA of fibroblasts in denatured collagen group was significantly higher than that in normal collagen group (P<0.01). Expressions of OB-cadherin of fibroblasts in denatured collagen, normal collagen, and common coverslips groups were respectively 5.17±0.28, 2.21±0.10, and 4.01±0.56. Expression of OB-cadherin of fibroblasts in denatured group was significantly higher than that in normal collagen group (P<0.01). (6) There was no significant difference in mRNA expression of collagen type Ⅰ of fibroblasts in denatured collagen, normal collagen, and common coverslips groups (F=2.71, P>0.05). The mRNA expressions of collagen type Ⅲ and α-SMA of fibroblasts in normal collagen group were significantly lower than those in denatured collagen group (P<0.01). Conclusions Denatured collagen type Ⅰ may influence the activity of fibroblasts, thus inducing fibroblasts differentiating into myofibroblasts.

Objective: To explore the application value of endoscope in probing the chronic wound with sinus tract in clinic. Methods: Twenty-eight chronic wounds with sinus tracts from 27 patients conforming to the inclusion criteria admitted to Outpatient Department of Wound Healing Center of Ruijin Hospital from December 2017 to March 2018 were investigated in a prospective and self-controlled trial. After being cleaned, the diameter of the opening of sinus tract was measured with a rule. A probe was used to measure the depth of a sinus tract according to the touch from the probe extremity in operation, and to measure the depth of a sinus tract that could be observed with naked eyes with the help of a pair of hemostatic forceps. Five minutes later, a probe was inserted deeply into the sinus tract to measure the depth under the endoscopic view combined with touch from the probe extremity in operation. Afterwards, the sinus tract was observed with endoscope, and the depth of the tract which could be observed under the endoscopic view was measured using a probe inserted deeply into the sinus tract. After completion of the above exploration, the sinus tract was infused with contrast agent Omnipaque 350 and scanned by computed tomography (CT) later to obtain its depth. The following indicators were calculated: the ratio of the depth of the sinus tract measured by CT to the diameter of the opening of the sinus tract (hereinafter referred to as the depth/diameter ratio of the sinus tract), the deviation rate comparing the depth of the sinus tract measured by conventional method (measured by probe only) and by endoscope (measured by probe under the endoscope view) with the depth of the sinus tract measured by CT (hereinafter referred to as the deviation rate of the measured depth of the sinus tract), the deviation rate comparing the depth of the sinus tract that could be observed measured by conventional method and by endoscope with the depth of the sinus tract measured by CT (hereinafter referred to as the deviation rate of the depth of the sinus tract that could be observed). Data were processed with paired t test. Pearson correlation analysis was applied to analyze the correlation between the depth/diameter ratio of the sinus tract and the deviation rate of the measured depth of the sinus tract and the deviation rate of the depth of the sinus tract that could be observed by conventional method and by endoscope. Results: The depth/diameter ratio of the sinus tract of this group of wounds was 1-32 (8±7). The deviation rate of the measured depth of the sinus tract and the deviation rate of the depth of the sinus tract that could be observed by conventional method were (19±14)% and (79±18)%, respectively, both obviously larger than (9±9)% and (25±25)% by endoscope (t=3.837, 13.626, P<0.01). Positive correlation existed between the depth/diameter ratio of the sinus tract and the deviation rate of the measured depth of the sinus tract by conventional method, and between the depth/diameter ratio of the sinus tract and the deviation rate of the depth of the sinus tract that could be observed by conventional method and by endoscope (r=0.514, 0.585, 0.651, P<0.01). However, there was no obvious correlation between the depth/diameter ratio of the sinus tract and the deviation rate of the measured depth of the sinus tract by endoscope (r=0.113, P>0.05). Conclusions Compared with the conventional method, application of endoscope is able to get more accurate data of chronic wounds with sinus tracts and observe the wounds with wider range.