Objective:To study the anatomic basis of superior trapezius myocutaneous and spina scapulae osteomyocutaneous flaps pedicled with transverse cervical vessel.Methods:The blood vessels,the size of the superior trapezius muscle and the spina scapulae were dissected and examined in 32 adult corpses.Results:The superior trapezius muscle was in the shape of trapezium.The border length (mm) of A,B,C and D was 174.63,157.18,86.98 and 80.95 in average respectively.The area of the muscle was 126.78 cm2 on the average.The spina scapulae was 131.21 mm in average length.The length (mm) of transverse cervical artery trunk, superficial cervical artery trunk and its ascending artery,spina scapulae branch artery was 42.50,27.80,43.12,28.75 in average,their external diameter(mm) was 2.71,2.39,1.96 and 0.50 respectively.Entering the muscle,the ascending artery had 3～6 branches with the external diameter of 0.5 mm or more.The venous vessels were following the copartner artery.Conclusion:The superior trapezius muscle and the spina scapulae may be made for the myocutaneous and osteomyocutaneous flaps pedicled with transverse cervical artery used in oral and maxillofacial reconstruction.

Objective: To investigate the effects of nerve growth factor(NGF) on the differentiation of goat bone marrow mesenchymal stem cells (BMSCs) into osteoblasts. Methods: The goat BMSCs were cultured in vitro and the marker proteins on the BMSCs surface were identified by flow cytometry. The third passage of BMSCs were randomly divided into blank control group, osteoblasts control group, NGF group and experimental group. The activities of alkaline phosphatase (ALP) and osteocalcin (0C) were examined and the of von Kossa staining method was used to observe the osteogenic differentiation. Results: CD90 and CD105 were strong positive while the CD34 and CD45 were negatively expressed in BMSCs. The activities of ALP and OC was significantly higher in the experimental group than that in the other three groups(P<0.05). The staining of von Kossa was positive and the black calcium nodules were appeared in the the osteoblasts control group. The number and the area of the calcium nodules were greater in the experimental group. But there were no significant differences of each index between the NGF group and the blank control group. Conclusion: NGF can′t induce goats BMSCs to osteoblasts, but can clearly promote the differentiation of goats BMSCs.

Objective To investigate the effects of apoptosis and related gene p21WAF1 and Bcl-2 in the tumorigenesis of primary gallbladder neoplasms. Methods p21WAF1 and Bcl-2 were detected by immunohistochemistry and apoptotic cells were stained in situ by terminal deoxynucleotidyl transferase mediated-dUTP nick end labeling(TUNAL) in 46 cases of primary gallbladder neoplasms with different histological grades. Results Apoptotic index and p21WAF1 were decreased,but the expression of Bcl-2 was increased with the increasing of the histological grades in primary gallbladder neoplasms (P

Objective To construct the eukaryotic expression vector of PRDX3 labeled with FLAG tag and to study its localization in human tongue cancer cell line SCC15.Methods PRDX3 gene was obtained from the breast library by PCR and cloned into PCDH vector to construct PCDH-FLAG-PRDX3.The plasmid was transiently transfected into 293T cells and the expression was detected by Western blot.Subcellular localization was detected by cellular immunofluorescence.Results The result of double digestion and sequencing showed that PCDH-FLAG-PRDX3 eukaryotic expression vector was constructed.The expression of FLAG-PRDX3 in human 293T cells was positively confirmed by Western blotting.In human tongue cancer cell line SCC15, the result of cellular immunofluorescence showed FLAG-PRDX3 was located in the cytoplasm rather than in the nucleus.Conclusion PRDX3 eukaryotic expression vector labeled with FLAG tag is constructed successfully, which is located in cytoplasm in human SCC15 cells.Construction and identification of PRDX3 could shed light on the function and mechanism of PRDX3 in tongue cancer.

Objective To investigate the influence of chondrocytes originating from different source on early chondrogenic differentiation of bone marrow mesenchymal stem cells (MSCs) in isolated co-culture system.Methods We applied hanging cell culture system to culture chondrocytes of different origin (osteoarthritis chondrocyte cells,nomal chondrocyte cells,infant chondrocyte cells) and controls.These chondrocytes and MSCs of the same origin were cultured in the common medium in a separated condition,and observed by microscope at 3,6,9,12 day after co culture.Expression levels of aggrecan,collagen type Ⅱ (Col 2),cartilage-specific transcription factor (Sox-9) in MSCs of different origin were determined by Real-time PCR.Results MSCs showed obviously morphological differentiation induced by chondrocytes of different origin at 12 day after coculture as compared with controls.Real-time PCR analysis showed that SOX9 mRNA level was stimulated by 1.7-fold,1.6-fold and 1.2-fold (all P＜0.05) and aggrecan mRNA level was increased by 2.8-fold,2.2-fold and 1.3-fold (all P＜0.05) in infant chondrocytes group,nomal chondrocytes group,osteoarthritis chondrocytes group respectively as compared with controls while COL2 mRNA level had no significant differences among the four groups.Corresponding protein signal level had obvious differences among the four groups,especially in infant chondrocytes as compared with osteoarthritis chondrocytes and nomal chondrocytes.Conclusions Isolated co-culture system may indirectly promote MSCs differentiation to chondrocytes by local micro-environment regulation.Chondrocytes of different origin have different effects on MSCs differentiation,but they could promote MSCs differentiation to chondrocytes.

Objective To construct eukaryotic expression vector of wild type E 4F1 and the mutant deleting amino acid region 32-81, and to detect the interaction between wild type or mutant E 4F1 and p53 and to study the effect of E4F1 on the expression level of p21.Methods Wild type and mutant sequences of E 4F1 were amplified from the mammary library using standard PCR and recombinant PCR .The sequences were cloned into pXJ 40-MYC vector to generate the MYC-E4F1 and MYC-E4F1(Δ32-81) recombinant plasmids that were transfected into 293T cells and identified by Western blotting . FLAG-p53 and MYC-E4F1 or MYC-E4F1(Δ32-81) were co-transfected into 293T cells and immunoprecipitation assay was performed to detect the interaction of wild type or mutant E 4F1 with p53.Wild type and mutant E4F1 expressing vec-tors were co-transfected into osteosarcoma U2OS cells and the expression of p21was detected.Results Recombinant plas-mids of MYC-E4F1 and MYC-E4F1(Δ32-81) were successfully constructed.Both wild type and mutant E4F1 interacted with p53.Deletion of amino acid region 32-81 of E4F1 increased the interaction .The expression level of p21 was in-creased by wild-type E4F1, but not by mutant E4F1.Conclusion The eukaryotic expression vector of wild type E4F1 and its deletion mutant is successfully constructed .Both of them interact with p53.Deletion of amino acid region 32-81 of E4F1 increases the interaction .This study contributes to further studies on the regulation and mechanism of E 4F1 on p53.

Objective To construct PES1 shRNA stable expression cell lines in tongue squamous cell carcinoma ( TSCC) cells and to study the effect of knockdown of PES 1 on the growth of TSCC cells .Methods Recombinant lentivirus carrying PES1 shRNA was packaged and obtained in 293T cells.TSCC cells (Tca8113, SCC6 and SCC15) were infected with the lentivirus and selected for stable cells .PES1 expression was identified by Western blot .The effect of inhibition of PES1 on the growth and cell cycle of TSCC cells was detected by growth curve and flow cytometry .Results TSCC cells stably expressing PES1 shRNA were constructed.Knockdown of PES1 inhibited cell proliferation and induced cell cycle ar-rest at G0/G1 phase.Knockdown of PES1 inhibited expression of cyclin D1 in TSCC cells.Conclusion Inhibition of PES1 results in reduced cell proliferation , cell cycle arrest at G 0/G1 phase and reduction of cyclin D 1 expression in TSCC cells . PES1 may be a target for TSCC gene therapy .

Objective To investigate the value of MR diffusion tensor imaging(DTI)in assessment of the microstructural changes of the trigeminal nerve,and analyze it's correlation with the degree of vascular compression. Methods Thirty-four patients with trigeminal neuralgia from November 2015 to April 2017 were retrospectively analyzed in this study.And they were treated by microvascular decompression(MVD). There were 11 cases of gradeⅠ,16 cases of gradeⅡand 7 cases of gradeⅢaccording to the severity of the contact between nerves and vessels during the operation. All of them were scanned with three dimensional time-of-flight(3D-TOF)sequences, three dimensional fast imaging employing steady state acquisition(3D-FIESTA)sequences and DTI before undergoing surgical decompression. According to the preoperative MR scans,the trigeminal nerves were divided into the healthy side without neurovascular contact (25 cases) and the healthy side with a neurovascular contact (9 cases).The DTI parameters of the trigeminal nerve,including the anisotropic fraction(FA)and the ADC values were obtained.Comparison of the FA and ADC values of the trigeminal nerve between the different stages of the affected side was performed with single factor analysis of variance, and the paired samples t test was used to compare the difference of FA and ADC values of bilateral trigeminal nerve. The difference of FA and ADC values between the asymptomatic side with or without vascular contact was compared with independent sample t test. Spearman correlation analysis was used to evaluate the correlation between DTI parameters and the degree of compression. Results The FA values of patients with grades Ⅰ,ⅡandⅢwere 0.311±0.009, 0.308±0.007 and 0.299±0.009 respectively,and there was significant difference among different levels(F=5.269,P<0.05).The ADC values of the three grades were(2.298 ± 0.309)×10-3,(2.214 ± 0.175)×10-3and (2.259 ± 0.248)×10-3mm2/s respectively, showing no statistically significant difference(F=0.402,P>0.05). The FA values of bilateral trigeminal nerves in healthy side without neurovascular contact and in healthy side with neurovascular contact were statistically significant (t=-32.528,-25.178,P<0.05). There was significant difference in the ADC value of bilateral trigeminal nerves in the group without neurovascular contact(t=2.162,P<0.05).There was no statistically significant difference in the ADC values of bilateral trigeminal nerves in the healthy side of the neurovascular contact group(P>0.05).There were no statistically significant differences in the FA and ADC values between the two groups on the healthy side of the trigeminal nerve(P>0.05).The FA value was negatively correlated with the degree of vascular compression (r=-0.453,P<0.05),while the ADC value was not correlated with the degree of vascular compression(P>0.05). Conclusion DTI imaging can be used to evaluate the degree of trigeminal nerve injury. More obvious vascular compression leads to lower FA value.

Objective:To investigate the clinical effect of laparoscopic assisted removal of greater omentum free transplantation combined with skin grafting for the repair of large area refractory wounds.Methods:From June 2013 to June 2018, 18 cases of lower extremity skin and soft tissue defects with multiple bone, joint, tendon and internal plants exposure were admitted to Hanzhong Central Hospital, including 12 males and 6 females, aged from 15 to 50 years old, with an average age of 32.6 years old. The area of skin and soft tissue defect: 30 cm×12 cm-53 cm×21 cm. The operation was divided into two stages. In the first stage, the greater omentum was acquired with the assist of laparoscope and free transplanted to cover the wound. After the greater omentum free transplantation was confirmed to survive, the split-thickness skin graft was applied for wound repair.Postoperative survival of the greater omentum and skin grafting, complications, appearance and function of lower limbs were observed and followed up.Results:The 18 operations were performed successfully, the area of omentum resection was 25 cm×10 cm-35 cm×15 cm, all the greater omentums survived after operation without complications such as intestinal adhesion, volvulus and peritonitis. The area of the skin grafting was 36 cm×8 cm-45 cm×22 cm. 16 cases skin grafting survived completely, 2 cases skin grafting were necrosis just local small area, and scar healed after dressing change. Postoperative follow-up of 6-12 months showed good appearance and function of lower limbs and satisfactory results.Conclusions:For the large area soft tissue defect wound of lower extremity, complicated with multiple deep tissues such as bone, joint and internal materials exposed, the greater omentum free transplantation under laparoscope combined with medium thick skin graft second stage has the advantages of good appearance and function after wound healing, less donor injury and fewer postoperative complications.

Objective:To investigate the clinical effect of laparoscopic assisted removal of greater omentum free transplantation combined with skin grafting for the repair of large area refractory wounds.Methods:From June 2013 to June 2018, 18 cases of lower extremity skin and soft tissue defects with multiple bone, joint, tendon and internal plants exposure were admitted to Hanzhong Central Hospital, including 12 males and 6 females, aged from 15 to 50 years old, with an average age of 32.6 years old. The area of skin and soft tissue defect: 30 cm×12 cm-53 cm×21 cm. The operation was divided into two stages. In the first stage, the greater omentum was acquired with the assist of laparoscope and free transplanted to cover the wound. After the greater omentum free transplantation was confirmed to survive, the split-thickness skin graft was applied for wound repair.Postoperative survival of the greater omentum and skin grafting, complications, appearance and function of lower limbs were observed and followed up.Results:The 18 operations were performed successfully, the area of omentum resection was 25 cm×10 cm-35 cm×15 cm, all the greater omentums survived after operation without complications such as intestinal adhesion, volvulus and peritonitis. The area of the skin grafting was 36 cm×8 cm-45 cm×22 cm. 16 cases skin grafting survived completely, 2 cases skin grafting were necrosis just local small area, and scar healed after dressing change. Postoperative follow-up of 6-12 months showed good appearance and function of lower limbs and satisfactory results.Conclusions:For the large area soft tissue defect wound of lower extremity, complicated with multiple deep tissues such as bone, joint and internal materials exposed, the greater omentum free transplantation under laparoscope combined with medium thick skin graft second stage has the advantages of good appearance and function after wound healing, less donor injury and fewer postoperative complications.