ObjectiveBased on the nuclear factor erythroid 2 related factor 2 （Nrf2）/heme oxygenase-1 （HO-1） signaling pathway， this paper explores the effect of Sinisan （SNS） on liver oxidative stress injury in cholestatic hepatitis rats and its mechanism. MethodThirty 6-week-old male SD rats were randomly divided into a control group， model group， low and high dose groups of SNS （2.5 and 5 g·kg^-1） and ursodeoxycholic acid group （UDCA， 63 mg·kg^-1）， with six rats in each group. Rats were administrated for seven consecutive days. On the 5th day， the control group was given olive oil of 10 mL·kg^-1， and the other groups were given alpha-naphthalene isothiocyanate （ANIT） of 80 mg·kg^-1. The serum biochemical indicator levels of cholestasis and the content of antioxidant factors in rat liver were detected by enzyme-linked immunosorbent assay （ELISA）. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in liver tissue. The relative mRNA and protein expressions of Nrf2， HO-1， and quinone oxidoreductase 1 （NQO1） in liver tissue were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultCompared with the control group， the model group showed a significant increase in the serum biochemical indicator levels of cholestasis and the content of antioxidant factors in liver tissue （P<0.01）. There were obvious pathological changes in the model group such as the disordered arrangement of hepatocytes， obvious congestion and necrosis in the portal area， infiltration of inflammatory cells， and destruction of the interlobular bile duct. The relative mRNA and protein expressions of Nrf2， HO-1， and NQO1 in liver tissue were significantly down-regulated in the model group （P<0.05， P<0.01）. Compared with the model group， the groups of SNS showed a significant decrease in the serum biochemical indicator levels of cholestasis and the content of antioxidant factors in liver tissue （P<0.01）， and the pathological liver injury was obviously improved. The necrotic area was reduced， and the infiltration of inflammatory cells was decreased. In addition， there was a small amount of extravasated blood in the interlobular vein. The relative mRNA and protein expressions of Nrf2， HO-1， and NQO1 in liver tissue were significantly up-regulated （P<0.05， P<0.01）. ConclusionSNS can significantly improve liver injury in cholestatic hepatitis rats， and its mechanism may be related to the inhibition of oxidative stress response mediated by the Nrf2/HO-1 signaling pathway.

Objective:To investigate the effects of fluoride exposure on kidney injury in rats and the sirtuin 3 (SIRT3)-fork head protein O3a (FOXO3a)-tensin homolog induced putative kinase 1 (PINK1)/E3 ubiquitin ligase (PARKIN) pathway.Methods:Twenty-four 4-week-old SD rats (clean grade, body mass 100 - 150 g) were selected and divided into three groups according to the randomized numeric table: control group, low fluoride group, and high fluoride group, with eight rats in each group (half male and half female). The control group was given free access to tap water (fluoride ion concentration < 0.5 mg/L), while the low fluoride and high fluoride groups were given free access to tap water and sodium fluoride solutions with fluoride ion concentrations of 5.0 and 50.0 mg/L, respectively, for a period of 180 days. The formation of dental fluorosis in rats was observed and recorded, and the femur, urine and blood samples of rats were collected to measure bone fluoride, urinary fluoride, and blood fluoride levels, and to detect kidney function related indicators (serum uric acid, creatinine, and urea nitrogen contents). Morphological changes of renal tissues stained with hematoxylin-eosin (HE) were observed under a light microscope. Real-time fluorescence quantitative PCR (qRT-PCR) and Western blotting were used to detect the mRNA and protein expression levels of renal SIRT3, FOXO3a, PINK1, PARKIN, microtubule associated protein 1 light chain 3 (LC3), autophagy receptor protein (P62), respectively.Results:Seven and one rats in the low and high fluoride groups were found to haveⅠdegree dental fluorosis, while zero and seven rats were found to haveⅡdegree dental fluorosis. Compared with the control group, rats in the low and high fluoride groups had higher levels of bone fluoride (μg/g: 1.18 ± 0.06, 2.16 ± 0.07 vs 0.52 ± 0.05), urinary fluoride (mg/L: 4.43 ± 0.11, 7.46 ± 0.09 vs 2.58 ± 0.14), blood fluoride (μg/ml: 0.77 ± 0.06, 1.68 ± 0.10 vs 0.52 ± 0.08), serum uric acid (μg/ml: 61.01 ± 4.17, 103.92 ± 5.43 vs 28.68 ± 2.91), creatinine (μg/ml: 74.82 ± 9.61, 132.05 ± 5.35 vs 22.38 ± 4.11), and urea nitrogen (μg/ml: 13.36 ± 1.27, 14.55 ± 0.34 vs 0.29 ± 0.07, P < 0.05). Under the light microscope, the kidneys of the control group showed tight and orderly arrangement of renal tubules and glomerular cells, with complete and clear cell contours. The low fluoride group was similar to the control group and no significant abnormalities were observed. The high fluoride group showed abnormal glomerular structure and atrophy, with some areas of renal tubules showing epithelial cell edema and unclear intercellular boundaries. The results of qRT-PCR assay showed that compared with the control group, the low and high fluoride groups had lower mRNA expression levels of SIRT3 (0.82 ± 0.03, 0.58 ± 0.02 vs 1.00 ± 0.08), P62 (0.75 ± 0.07, 0.28 ± 0.09 vs 1.00 ± 0.07, P < 0.05), and higher mRNA expression levels of FOXO3a (1.35 ± 0.04, 3.01 ± 0.23 vs 1.00 ± 0.08), PINK1 (1.58 ± 0.09, 3.28 ± 0.09 vs 1.00 ± 0.07), PARKIN (1.51 ± 0.04, 1.67 ± 0.10 vs 1.00 ± 0.05), LC3 (1.74 ± 0.07, 2.38 ± 0.18 vs 1.00 ± 0.08, P < 0.05). The results of Western blotting showed that compared with the control group, the low and high fluoride groups had lower protein expression levels of SIRT3 (0.91 ± 0.01, 0.55 ± 0.03 vs 1.00 ± 0.01), P62 (0.94 ± 0.27, 0.66 ± 0.38 vs 1.00 ± 0.19, P < 0.05), and higher protein expression levels of FOXO3a (1.14 ± 0.03, 1.22 ± 0.05 vs 1.00 ± 0.02), PINK1 (1.46 ± 0.03, 1.56 ± 0.03 vs 1.00 ± 0.05), PARKIN (1.98 ± 0.02, 2.33 ± 0.11 vs 1.00 ± 0.06), LC3 (4.10 ± 0.58, 4.93 ± 0.33 vs 1.00 ± 0.13, P < 0.05). Conclusion:Exposure to fluoride can cause renal tissue injury in rats, with downregulation of SIRT3 and P62 expression levels, and upregulation of FOXO3a, PINK1, PARKIN, and LC3 expression levels.

OBJECTIVE: To investigate the feasibility and effectiveness of bilateral facial perforator artery flap in repairing large area defect in middle and lower part of nose. METHODS: The clinical data of 18 patients with large area defect in middle and lower part of nose repaired by bilateral facial perforator artery flap between January 2019 and December 2022 were retrospectively analyzed. Among them, there were 13 males and 5 females, the age ranged from 43 to 81 years, with an average of 63 years. There were 3 cases of nasal trauma, 4 cases of basal cell carcinoma, 8 cases of squamous cell carcinoma, 1 case of lymphoma, and 2 cases of large area solar keratosis. The size of the defect ranged from 3.0 cm×3.0 cm to 4.5 cm×4.0 cm; the size of unilateral flap ranged from 3.0 cm×1.3 cm to 3.5 cm×2.0 cm, and the size of bilateral flaps ranged from 3.3 cm×2.6 cm to 4.5 cm×4.0 cm. RESULTS: One patient developed skin flap necrosis after operation, and a frontal skin flap was used to repair the wound; 1 case gradually improved after removing some sutures due to venous congestion in the skin flap, and the wound healing was delayed after dressing change; the remaining 16 cases of bilateral facial perforator artery flaps survived well and all wounds healed by first intention, without any "cat ear" malformation. All 18 patients had first intention healing in the donor area, leaving linear scars without obvious scar hyperplasia, and no facial organ displacement. All patients were followed up 3-12 months, with an average of 6 months. Due to the appropriate thickness of the flap, none of the 18 patients underwent secondary flap thinning surgery. All flaps had good blood circulation, similar texture and color to surrounding tissues, symmetrical bilateral nasolabial sulcus, and high patient satisfaction. CONCLUSION The bilateral facial perforator artery flaps for repairing large area defect in middle and lower part of nose can achieve good appearance and function, and the operation is relatively simple, with high patient satisfaction.
Male ; Female ; Humans ; Adult ; Middle Aged ; Aged ; Aged, 80 and over ; Plastic Surgery Procedures ; Skin Transplantation ; Retrospective Studies ; Soft Tissue Injuries/surgery* ; Perforator Flap/blood supply* ; Arteries/surgery* ; Cicatrix/surgery* ; Treatment Outcome ; Skin Neoplasms/surgery*

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.

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 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 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 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.