Objective To investigate the distribution of intramuscular nerves in the different skeletal muscles of the rabbit. Methods Muscle architecture and modified sihler's neural staining methods were used. Results The nerve branches of innervating flat M.pectoralis major arose from the anterior pectoral and posterior pectoral nerve.The former innervated cross fibers mainly,it penetrated the middle of muscle belly and formed a “U-Shaped” nerve ansa.The latter innervated oblique fibers mainly.There were many anastomoses between them.The nerve of pennate M.plantaris derived from tibial nerve.After entering muscle,the nerve trunk gradually divided into many primary branches toward medial and lateral fibers,these branches then subdivided into numerous arboroid second and terminal branches toward all of muscle fibres;The nerve of innervating spindle M.extensor digitorum longus came from N.fibularis communis.Two extramuscular nerve trunk were seen.Superior trunk mainly innervated those muscle fibers of inserting in the second toe,Inferior one mainly distributed to the fibers of rest within this muscle.Conclusion The spatial arrangement of the muscle fibres were related to distribution of the intramuscular nerves;Walking of intramuscular nerve branches had two patterns which run perpendicular and/or parallel to muscle fasciculi.

Objective To investigate the relationship between the methylation of CpG island of RUNX3 gene promoter and its expression in a human cutaneous malignant melanoma cell line A375, and to assess the role of RUNX3 gene methylation in the pathogenesis of human cutaneous malignant melanoma. Methods Cultured A375 cells were treated with various concentrations (0, 1, 5, 10, 20 μmol/l) of 5-azacyti-dine for 24 or 72 hours followed by another 5 days of culture. Then, methylation-specific PCR (MSP) was performed to evaluate the methylation status of RUNX3 promoter region, and Western-blot analysis to detect the protein expression of RUNX3 in A375 cells. Results The RUNX3 gene promoter region was hypermethylated in untreated A375 cells, along with the absence of protein expression of RUNX3. However, after the treatment with 5-azacytidine, the promoter region of RUNX3 gene was demethylated partly, and the expression of RUNX3 protein was restored in A375 cells. Further, the expression intensity was directly correlated with the concentration of 5-azacytidine. Conclusions The promoter hypermethylation of RUNX3 gene may be related to the silencing of RUNX3 gene expression in A375 cells, whereas 5-azacytidine can cause the demethylation of RUNX3 gene, reactivate the gene which has been inactivated by the promoter hypermethylation, and finally induce the re-expression of RUNX3 protein.

Objective To investigate the effect of phorbol-12-myristate-13-acetate(PMA)on cyclooxygenase-2(COX-2) mRNA and protein expression in cultured human HaCaT keratinocytes,and the mechanism for cytotoxity of PMA against keratinocytes.MethodsRT-PCR and Westem blot were utilized to detect the expression of COX-2 mRNA and protein in cultured HaCaT ells at 24 hours after the treatment with various concentrations of PMA (0.1,1.0,10 mg/L).ResultsWithout any treatment,there was no or a weak expression df COX-2 mRNA and protein in HaCaT cells;incubation witll PMA resulted in the induction of the expression of COX-2 in HaCaT cells.The expression levels of COX-2 mRNA and protein in 10 mg/L PMA-pretreated HaCaT cells were significantly higher than those in 1.0 mg/L PMA-pretreated HaCaT cells,which was in turn higher than that in 0.1 mg/L PMA-pretreated cells and untreated cells;the difrerence was statistically significant (all P＜0.01).Conclusion These results suggest that PMA may be involved in keratinocyte tumorigenesis by upregulating he expression of COX-2 as well as synthesis and release of prostaglandin in keratinocytes.

Objective To evaluate the clinical efficacy and safety of combined ozone hydrotherapy for the treatment of atopic dermatitis(AD). Methods A total of 60 patients with moderate or severe AD aged from 6 to 65 years were enrolled, and randomly and equally divided into a test group and a control group. Both the two groups were treated with oral levocetirizine capsules 5 mg once a day, topical tacrolimus ointment twice a day, and topical moisturizers. The test group was additionally treated with ozone hydrotherapy 3- 5 times every week. The treatment lasted 2 weeks. The severity scoring of atopic dermatitis (SCORAD) score, visual analog scale (VAS) score, dermatology life quality index (DLQI) or children′s dermatology life quality index (CDLQI) score were assessed before and after the treatment, and compared between the two groups. Enzyme?linked immunosorbent assay(ELISA) was performed to measure the levels of interleukin?4(IL?4)and nerve growth factor(NGF)in peripheral blood from the patients before and after the treatment. Results After 2?week treatment, the SCORAD scores, VAS scores and DLQI/CDLQI scores significantly decreased from 42.13 ± 16.03, 7.14 ± 2.12 and 14.92 ± 5.94 before the treatment to 27.3 ± 11.01, 2.23 ± 1.31 and 9.69 ± 4.17 respectively in the test group(all P<0.05), and from 43.36 ± 17.78, 6.45 ± 1.29 and 15.15 ± 5.40 to 39.65 ± 16.67, 3.32 ± 0.87 and 12.84 ± 5.37 respectively in the control group(all P<0.05). In addition, all the three scores were significantly lower in the test group than in the control group after 2?week treatment(all P<0.05). A significant decline was also observed in the levels of IL?4 and NGF in peripheral blood in the two groups at the end of the treatment compared with those at baseline(all P<0.05), and the magnitude of decline in IL?4 levels was significantly higher in the test group than in the control group(t=8.57, P<0.05), while that in NGF levels was insignificantly different between the two groups(t=9.51, P>0.05). Conclusion Combined ozone hydrotherapy can effectively and safely improve the condition of patients with AD, likely by decreasing the levels of IL?4 in peripheral blood.

OBJECTIVEThe effect of triptolide on the DNA methylation level of MMP-9 gene and the mRNA expression of tissue inhibitors of met-alloproteinases (TIMPs) were examined in human fibrosarcoma HT-1080 cells to explore the molecular mechanisms involved in the anticancer activity of triptolide.

METHODHT-1080 cells were cultured in MEM containing 10% newborn calf serum and 1% penicillin-streptomycin. Triptolide was dissolved in dimethyl sulfoxide (DMSO) at a concentration of 1 goL-1 and stored at -20 degrees C. Triptolide was freshly diluted with culture medium perior to use and directly added to cell cultures at the indicated concentration, and incubated for 72 hours at 37 degrees C in a humidified atmosphere with 5% CO2, with changes of reagents every 24 hours. Methylation specific PCR (MSP)was applied to assess the methylation status of MMP-9 gene promoter, and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was employed to measure the mRNA expression of tissue inhibitors of metalloproteinases (TIMPs) in human fibrosarcoma HT-1080 cells after 72 hours of treatment with 6 nmol x L(-1), 12 nmol x L(-1) or 18 nmol x L(-1) triptolide, respectively.

RESULTSThe methylation index of MMP-9 gene promoter was statistically elevated in HT-1080 cells after 72 hours of treatment with 18 nmol L(-1) triptolide, compared with those in controls (0.61 +/- 0.10 vs 0.39 +/- 0.10, P < 0.05), while no significant difference was noted between 6 nmol x L(-1) or 12 nmol x L(-1) triptolide treated HT-1080 cells and controls (0.40 +/- 0.15 vs 0.39 +/- 0.10, 0.46 +/- 0.20 vs 0.39 +/- 0.10, respectively, both P > 0.05). The mRNA expression of TIMP-1, -2, -3 or -4 was not significantly changed in HT-1080 cells after 72 hours of treatment with the indicated concentrations of triptolide, respectively compared with those in controls (all P > 0.05).

CONCLUSIONThe results demonstrated that triptolide upregulates the methylation level of MMP-9 gene in HT-1080 cells in vitro.

Antineoplastic Agents, Alkylating ; pharmacology ; Cell Line, Tumor ; DNA Methylation ; drug effects ; Diterpenes ; pharmacology ; Epoxy Compounds ; pharmacology ; Fibrosarcoma ; drug therapy ; genetics ; metabolism ; Gene Expression Regulation, Enzymologic ; drug effects ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Phenanthrenes ; pharmacology ; Tissue Inhibitor of Metalloproteinases ; genetics ; metabolism

Objective:To investigate the preoperative anxiety level of patients with mouth and maxillofacial neoplasms and analyze the influencing factors.Methods:Using the convenient sampling method, a total of 385 patients with mouth and maxillofacial neoplasms who planned to receive surgery in a ClassⅢ Grade A stomatological hospital in Tianjin from June 2018 to January 2019 were selected as the research objects. General information questionnaire for patients, Self-Rating Anxiety Scale (SAS) , Social Support Rating Scale (SSRS) and Trait Coping Style Questionnaire (TCSQ) were used to investigate patients' general information, anxiety state, social support and coping style.Results:Totally 385 questionnaires were distributed, 361 questionnaires were recovered. In this study, 125 patients (34.63%) had anxiety symptoms, including 105 patients with mild anxiety (29.09%) , 18 patients with moderate anxiety (4.99%) , and 2 patients with severe anxiety (0.55%) . The results of multiple stepwise regression showed that active coping style, diagnosis, social support, occupation, payment method of hospitalization expenses and gender were the main influencing factors of preoperative anxiety in patients with mouth and maxillofacial tumors, which jointly explained 43.26% of the variation ( P<0.05) . Conclusions:The incidence of preoperative anxiety in patients with mouth and maxillofacial neoplasms is relatively high, especially for female patients with less social support, relatively negative coping style and diagnosis with malignant tumor. Medical staff should give personalized psychological intervention and health education in time to help patients successfully pass the perioperative period.

OBJECTIVE: To evaluate the short-term efficacy and safety of propranolol for problematic infantile hemangiomas. METHODS: Oral propranolol was administered to 68 infants with heamngiomas diagnosed by clinical evaluation and adjuvant examination at 1.0~2.0 mg per kilogram of body weight per day, divided to 2 or 3 times. The patients revisited once a month. The changes of the tumor size, texture, and color were monitored and recorded at a regular interval.The adverse effects after medication were observed and managed accordingly.The short-term results were evaluated using a 4-grade system. RESULTS: All the 68 infants were followed up for 3-13 months, except that 1 infants combined with other diseases and 4 withdrew.The overall response was Scale 1 in 8 infants, Scale II in 13, Scale III in 29, and Scale IV in 13. No serious adverse effects were seen, but none cured entirely as well. CONCLUSION Oral propranolol is safe and effective for infantile heamngioma with good short-term result. It could be used as the primary drug for problematic infantile hemangiomas at the rapid growth stage of hemangiomas.
Female ; Hemangioma ; drug therapy ; Humans ; Infant ; Male ; Propranolol ; therapeutic use ; Prospective Studies ; Skin Neoplasms ; drug therapy ; Subcutaneous Tissue

Objective:To identify the pathogenic gene of a pedigree with symphalangism and to prove the pathogenicity of this locus in vitro.Methods:The clinical data of patients’families were collected at Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, peripheral blood was collected and genomic DNA was extracted and NOG, FGF9, GDF5 exon regions were amplified by PCR, and the exon gene mutations were detected by first-generation sequencing technique. The structure of noggin-GDF5 protein complex was simulated in silicon. COS-7 cells were transfected with 5 μg empty plasmid, wild type plasmid and V202G mutant plasmid in vitro. Each group of plasmids was transfected into 3 well cells. The experiment was repeated for 3 times, and the expression of noggin protein was detected by Western blotting. C2C12 cells were also transfected with the above plasmids in vitro for osteogenic differentiation. By applying alkaline phosphatase staining and quantitative assay. Relative expression level of osteoblast-related genes Col1α1, ALP and Runx2 were detected by qRT-PCR. Each group of plasmids was transfected into 3 well cells, and the experiment was repeated for 3 times. All statistical analysis were performed by Prism 6 software. The result were shown as mean±standard deviation, and the comparison between groups was done by unpaired t-test. Data were considered statistically significant when P value is less than 0.05. Results:Both the proband and his mother suffered from symphalangism. The result of Sanger sequencing showed that there was a heterozygous missense mutation of NOG gene (p.V202G) in all patients in this pedigree. No disease-related mutations were detected in FGF9 and GDF5. Computer three-dimensional mechanism simulation showed that the site was located at the α helix. The result of Western blotting showed that the expression of mutant protein was significantly lower than that of wild type. Osteogenic differentiation in vitro showed that the inhibitory effect of V202G mutant protein on osteogenic differentiation decreased. The quantitative result of alkaline phosphatase staining showed that the alkaline phosphatase activity in the vector group was (12.3±0.8) U/L, and the alkaline phosphatase activity in the wild type plasmid group was (2.6±0.3) U/L, which was significantly lower than that in the vector group ( t=11.550, P<0.001). The alkaline phosphatase activity in the mutant plasmid group was (10.8±0.3) U/L. There was no significant difference between the mutant group and the vector group ( t=1.830, P=0.141). The mRNA expression level of osteogenesis-related genes was consistent with the above result . Compared with vector group, the expression of osteogenesis-related genes in wild-type noggin group decreased significantly ALP、 Col1α1 and Runx2 ( t=5.987, 4.498, 4.170; P=0.004, 0.011, 0.014). There was no significant difference between mutant plasmid group and blank vector group in ALP、 Col1α1 and Runx2 ( t=0.433, 0.177, 1.159; P=0.688, 0.868, 0.311). Conclusions:NOG gene c. 605T < G p. V202G is a novel mutation in symphalangism, which is located in the α helix of noggin protein, leading to the decrease of the expression of noggin protein and the manifestation of ankylosis.

Objective:To identify the pathogenic gene of a pedigree with symphalangism and to prove the pathogenicity of this locus in vitro.Methods:The clinical data of patients’families were collected at Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, peripheral blood was collected and genomic DNA was extracted and NOG, FGF9, GDF5 exon regions were amplified by PCR, and the exon gene mutations were detected by first-generation sequencing technique. The structure of noggin-GDF5 protein complex was simulated in silicon. COS-7 cells were transfected with 5 μg empty plasmid, wild type plasmid and V202G mutant plasmid in vitro. Each group of plasmids was transfected into 3 well cells. The experiment was repeated for 3 times, and the expression of noggin protein was detected by Western blotting. C2C12 cells were also transfected with the above plasmids in vitro for osteogenic differentiation. By applying alkaline phosphatase staining and quantitative assay. Relative expression level of osteoblast-related genes Col1α1, ALP and Runx2 were detected by qRT-PCR. Each group of plasmids was transfected into 3 well cells, and the experiment was repeated for 3 times. All statistical analysis were performed by Prism 6 software. The result were shown as mean±standard deviation, and the comparison between groups was done by unpaired t-test. Data were considered statistically significant when P value is less than 0.05. Results:Both the proband and his mother suffered from symphalangism. The result of Sanger sequencing showed that there was a heterozygous missense mutation of NOG gene (p.V202G) in all patients in this pedigree. No disease-related mutations were detected in FGF9 and GDF5. Computer three-dimensional mechanism simulation showed that the site was located at the α helix. The result of Western blotting showed that the expression of mutant protein was significantly lower than that of wild type. Osteogenic differentiation in vitro showed that the inhibitory effect of V202G mutant protein on osteogenic differentiation decreased. The quantitative result of alkaline phosphatase staining showed that the alkaline phosphatase activity in the vector group was (12.3±0.8) U/L, and the alkaline phosphatase activity in the wild type plasmid group was (2.6±0.3) U/L, which was significantly lower than that in the vector group ( t=11.550, P<0.001). The alkaline phosphatase activity in the mutant plasmid group was (10.8±0.3) U/L. There was no significant difference between the mutant group and the vector group ( t=1.830, P=0.141). The mRNA expression level of osteogenesis-related genes was consistent with the above result . Compared with vector group, the expression of osteogenesis-related genes in wild-type noggin group decreased significantly ALP、 Col1α1 and Runx2 ( t=5.987, 4.498, 4.170; P=0.004, 0.011, 0.014). There was no significant difference between mutant plasmid group and blank vector group in ALP、 Col1α1 and Runx2 ( t=0.433, 0.177, 1.159; P=0.688, 0.868, 0.311). Conclusions:NOG gene c. 605T < G p. V202G is a novel mutation in symphalangism, which is located in the α helix of noggin protein, leading to the decrease of the expression of noggin protein and the manifestation of ankylosis.

As a secondary preventive measure for patients with coronary heart disease, cardiac rehabilitation can improve their quality of life and reduce mortality. One key aspect is to ensure that patients receive sufficient information. This article provides a review of the content, influencing factors, and support strategies for the cardiac rehabilitation information needs of patients with coronary heart disease, in order to provide reference for establishing a secondary prevention strategy for patients with coronary heart disease guided by information needs.