BACKGROUND:Fracture healing in diabetic patients is usual y unsatisfactory because of hormones and metabolic disorder, and an eventual multiple organ dysfunction resulting from high blood glucose. OBJECTIVE:To dynamical y observe the changes of cytokines during the fracture healing process in diabetic rats before and after insulin treatment. METHODS:A total of 120 Sprague-Dawley rats were included in this study. Of them, 90 rats intravenously injected with 5%tetraoxypyrimidine to induce rat models of diabetes were randomized into insulin treatment and diabetes groups, respectively. The remaining 30 rats were intravenously injected with equal volume of saline and selected as control group. The next day, blood glucose was determined. Healing at 1, 4, and 8 weeks after fracture were observed by the X-ray film. Biomechanical strength of the injured right tibia was measured at 4, 6, and 8 weeks after modeling. Cytokines in the osteotylus were determined by immunohistochemical staining and in situ hybridization technique. RESULTS AND CONCLUSION:The X-ray films showed that the speed of fracture healing in the diabetes group was slower than insulin treatment and control groups. Biomechanical strength of the osteotylus in the diabetes group was significantly decreased compared with the insulin treatment and control groups. However, there were no significant differences in above-mentioned parameters between the control and insulin treatment groups. Bone morphogenetic protein 2, basic fibroblast growth factor, transforming growth factor-beta, and vascular endothelial growth factor were widely expressed in the osteotylus and their expressions in diabetes group were significantly lower and slower than those in the control and insulin treatment groups. There was no statistical difference between control and insulin treatment groups. These results indicate that osteotylus formation speed, biomechanical strength, and growth factor expressions at the fracture site in diabetes rats were decreased compared with normal rats. Insulin treatment can enhance cytokine levels at the fracture site, thereby promoting the osteoblast proliferation and fracture healing.

Objective To establish a culture protocol for Th17 cells in vitro and evaluate the effect of curcumin on the differentiation and functions of Th17 cells and explore the related mechanisms. Methods Splenic CD4+CD25- T cells of C57BL/6 mice were isolated and purified with magnetic bead methods, and were co-cultured with plate-bound anti-CD3 and anti-CD28 antibodies and with transforming growth factor (TGF)-β, interleukin (IL)-6, IL-23, anti-interferon-γ antibody and anti-IL-2 antibody for Th17-polarization. The cultured Th17 cells were allocated into four groups: the control group, in which T cells were cultured on the basis of the above protocol; the low-concentration curcumin (CM-L, 5 μmol/L) and high-concentration (CM-H, 25 μmol/L) curcumin groups, and sirolimus (SRL, 100 ng/ml) group. The proportion of Th17 cells was detected with flow cytometry, and the mRNA expression of retinoic acid-related orphan receptor (ROR)γt, IL-17A, IL-21 was examined with real-time quantitative polymerase chain reaction. Finally, the protein expression of RORγt and the phosphorylation levels of signal transducer and activator of transcription (STAT) 3 were measured using western blotting analysis. Results The proportion of Th17 cells in the freshly isolated CD4+CD25- T cells was (3.1 ±0.4)%, whereas the proportion of the cells cultured with the protocol could get [(54.1±3.4)%, P<0.01]. As compared with the control group, the Th17 cells proportion in CM-L, CM-H and SRL groups was significantly decreased [CM-L (40.3±2.8)%, CM-H (25.8±2.3)%, SRL (25.0±2.0)%versus the control (54.1±3.4)%, t=6.15, 12.63, 12.97, P<0.01], and no marked difference was seen between CM-H group and SRL group (ft>0.05). Moreover, the mRNA levels of RORγt, IL-17A and IL-21, the protein expression of RORγt and the phosphorylation levels of STAT 3 in CM-L, CM-H and SRL groups were also lower than those in the control group (IL-17A mRNA: CM-L 0.81±0.05, CM-H 0.61±0.05, SRL 0.58±0.05, Control 1.01 ±0.11, t=4.81, 8.52, 8.89; IL-21 mRNA: CM-L 0.73±0.06, CM-H 0.49±0.03,SRL 0.59±0.03, Control 1.12±0.11, t=5.98, 9.22, 7.95, P<0.01). The mRNA level of IL-21 in the CM-H group was lower than that in the SRL group (P<0.05). Conclusion In vitro, curcumin can inhibit the differentiaton of splenic CD4+CD25- T cells into Th17 cells in the setting of Th 17-polarization and inhibit the expression of IL-17A and IL-21 mRNA, which is associated with the inhibitive effect of curcumin on RORγt expression and STAT 3 phosphorylation.

objective To explore a new method of repairing skin defects complicated with fracture and tendon rupture at the middle and distal sections of the second to fifth fingers. Methods The reversed dorsal metacarpal island flap was designed to be pedicled on the digital proper artery-common digital artery-fingerweb artery-dorsal metacarpal artery-cutaneous branch of dorsal metacarpal artery.In repairing digital palmar skin defects,after the flap was dissected,the proximal incision was extended along the direction of dorsal metacarpal nerve to harvest an enough length of the nerve so that the dorsal metacrppal nerve can be anastomosed with the digital proper nerve to restore the sensation of finger pulp.From the June 2003 to March 2009,the flap was used to repair 26 fingers in 24 patients with middle and distal digital skin defects complicated with fracture and tendon rupture.They were 17 men and 7 women,aged from 16 to 63 years (average,37 years).There were 15 cases of palmar skin defect and 9 cases of dorsal skin defect.In the 2 cases of combined tendon defects,a section of the extensor tendon of index(or little) finger was dissected together with the flap to repair the tendon rupture. Results The areas of the flap ranged from 3.1 cm ×1.6 cm to 6.0 cm × 4.0 cm.The flaps survived in all 24 cases without any vascular crisis.Twenty-two patients obtained an average follow-up of 14 months (from 4 to 32 months) but 2 were lost to the follow-up.The flaps were fine in texture,colour and appearance.The finger pulps appeared full and recovered sensations of pain and temperature.The average two-point discrimination was 7.5 mm (from 5 to 9 mm).Sensory function evaluation revealed an outcome of S3 + ～ S4.Tendon adhesion occurred in 4 cases which recovered digital function following secondary lysis 3 to 6 months postoperation. Conclusion Application of the reversed dorsal metacarpal island flap pedicled on the digital proper artery is a good way to repair skin defects complicated with fracture and tendon rupture at the middle and distal sections of the second to fifth fingers.

Objective To investigate the mechanism of drug resistance mediated by micF gene and outer membrane porin F ( OmpF) in Shigella strains. Methods Shigella strains were isolated from stool samples of patients who presented to the Second Hospital of Tianjin Medical University with acute diar-rhea in 2015. Antibiotic susceptibility test was performed to screen out the multidrug-resistant and non-multi-drug-resistant strains. The ompF gene was amplified by PCR. The micF and ompF genes at transcriptional levels in the two groups of strains were detected by quantitative real-time RT-PCR. Intracellular concentra-tions of ciprofloxacin in the two groups of Shigella strains were measured by automatic microplate reader. Re-sults According to the result of antibiotic susceptibility test, 13 strains that were resistant to 3 or more than 3 antibiotics were classified into the multidrug-resistant group, while the other 8 strains that were sensitive to all antibiotics used in this study or only resistant to 1 or 2 antibiotic were classified into the non-multidrug-re-sistant group. All of the 21 Shigella strains carried the ompF gene. Compared with the non-multidrug-resist-ant strains, the multidrug-resistant strains showed higher expression of micF gene, but lower expression of ompF gene. The differences in micF and ompF genes between the two groups were statistically significant. The result of correlation analysis suggested that there was a negative correlation between micF and ompF genes (r=-0. 244). The intracellular concentrations of ciprofloxacin in multidrug-resistant strains were low-er than those in the non-multidrug-resistant strains (P<0. 001). Conclusion The decreased expression of OmpF was one of the possible mechanisms of multidrug-resistance in Shigella strains. The micF gene was negatively related to the expression of OmpF. Moreover, the decreased intracellular concentrations of cipro-floxacin in multidrug-resistant strains might be related to the decreased expression of OmpF.

Objective To investigate the feasibility of repairing soft-tissue defects of the fifth phalange and the back of hand with ulnar palmar artery perforator flaps from the little finger.Methods Based on anatomic dissection,the fifth phalange ulnar palmar artery perforator flaps were created and transferred to repair soft-tissue defects at the little finger and the back of hand in 15 cases.Types of injury were stamping injury in 5 cases,planer injury in 4 cases,mechanical crash injury in 3 cases,blast injury in 2 cases,and cicatrical contracture following electric burn in 1 case.Injury involved in the palmar aspect near the middle segment of fifth phalange in 4 cases,dorsal aspect near the middle segment of fifth phalange in 6 cases,ulnar mesiodistal of the back of hand in 3 cases,and distal ulnar palmar aspect of hands in 2 cases.There were 6 patients wounded in left hands and 9 patients wounded in right hands.Results All flaps survived and all wounds healed by first intention.At the follow-up of 2-18 months,the flaps resurfaced the soft-tissue defects with good color and texture match and the maintenance of contour and function of donor and recipient sites were satisfactory.Conclusion The fifth phalange ulnar palmar artery perforator flap,as it has advantages of constant perforator vessels,rich blood supply and good texture and can be operated safely and easily,is considered an ideal treatment choice in repairing softtissue defects of the fifth phalange and the back of hand.

Objective To explore the methods for endoscopy nasal microsurgery and multiple techniques in treatment of nasopharyngeal angiofibromas. Methods CT, MRI and digial substraction angiography (DSA) and endoscopic examination had been used. ALL 12 patients accepted preoperative feeding artery embolum, hypotention anesthesia and injected adrenaline-lidocaine. Microelectrotomy-electrocoagulation had been used to remove tumor. Results All patients were treated by endoscopic nasal surgery. No complications had occurred. No recurrence were found in follow-up period. Conclusion The important factors to reduce amount of introperative bleeding are to select proper operative patients and use multiple techniques. Endoscopy nasal surgery has dominance in reducing complications, wound, tumor recurrence, operative time and nasal function.

Objective To investigate the effect of curcumin on autophagy and inflammatory response in mice subject to renal ischemia reperfusion.Method Forty male C57BL/6 mice were randomly divided into four groups (n =10 each):sham-operated (SO) group (Abdominal incision was made to expose the kidneys,bilateral renal pedicle dissociated and the abdomen sewed),ischemia reperfusion (IR) group,curcumin (CM) group (given CM 10 mg/kg) and 3-methyladenine (3-MA) group (given 3-MA 15 mg/kg).Six and 24 h after reperfusion,renal function was tested by determining the serum creatinine (Scr) and blood urea nitrogen (BUN) levels,and the morphological changes in the kidney tissue were observed.The expression of LC3,Beclin 1,Rab7 and LAMP2 in kidney tissue was detected by fluorescence quantitative RT-PCR and Western blotting.Fluorescence quantitative reverse transcription PCR and enzyme-linked immunosorbent assay were used to examine the expression of IL-6,IL-10,IL-17 and TNF-α in kidney tissue and serum.Result As compared with IR group and 3-MA group,the Scr and BUN levels in the CM group were significantly decreased (P＜0.01),and the renal morphological changes were improved significantly (P＜0.01).The mRNA and protein expression of LC3,Beclin 1,Rab7 and LAMP2 was significantly increased in kidney tissue,and the expression of IL-6,IL-17 and TNF-α was reduced,while IL-10 was increased (P＜ 0.01) in the CM group as compared with IR group and 3-MA group.Conclusion Curcumin possesses a protective effect against renal ischemia reperfusion injury in mice,which is probably mediated by promoting autophagy and subsequently inhibiting inflammatory response.

Objective To observe the effect of human umbilical cord-derived mesenchymal stem cells (hMSCs) on type 2 diabetic rats, and to explore the possible mechanism. Methods Type 2 diabetic rats were induced by high-fat diet combined with a low dosage of streptozotocin ( STZ, 25 mg/ kg). After 3 × 106 hMSCs suspended in 1 ml PBS or 1ml 10-fold concentrated hMSC supernatant were intravenously infused into the rats via the tail vein, the blood glucose levels were measured every day. One week later, intraperitoneal glucose tolerance test and insulin tolerance test were performed to evaluate the effects of hMSCs on diabetic rats. Pancreatic tissues were collected for insulin/ glucagon immunofluorescence staining. Results After hMSCs infusion, blood glucose level and homeostasis model of assessment for insulin resistance index were significantly decreased in type 2 diabetic rats(both P<0. 01). The glucose tolerance and insulin tolerance were greatly alleviated by hMSCs(all P<0. 01). Intravenously infused 1ml 10-fold concentrated hMSC supernatant showed a similar result to hMSCs. Conclusion In type 2 diabetic rats, hMSCs are able to effectively lower the blood glucose level, improve insulin sensitivity, and increase the number of β cells, which seems to be mediated by their secreted molecules.

Objective To investigate functional role and clinical significance of the methylation in the promoter of TPAP gene in the development and progression of pancreatic cancer.Methods Surgically resected specimen from 68 patients who were pathologically diagnosed as pancreatic cancer in Changhai Hospital from July 2006 to August 2009 were collected.The methylation in the promoter of TPAP gene in tumor and nontumor adjacent tissue was detected by methylation specific PCR.Results The methylation rate of tumor and non-tumor adjacent tissue was (0.214 ± 0.057) ％ and (0.084 ± 0.096) ％,respectively,and pancreatic cancer tissue had significantly higher methylation rate than the adjacent tissue.Hypermethylation of TPAP gene was not correlated with age,gender,tumor differentiation,lymphatic metastasis,serum CEA and CA19-9,but was positively correlated with distant metastasis.Conclusions Hypermethylation in the promoter of TPAP gene may participate in the invasion and metastasis of pancreatic cancer and the hypermethylation of the promoter is closely associated with the tumorigenesis and development of pancreatic cancer.

Objective To study the expression difference of the insulin-like growth factor-I (IGF-I)and IGF-binding proteins-3,4,5(IGFBP-3,4,5)in the peripheral and local areas in senile hip fracture and discuss the correlation between the expression and the fracture types. Methods The study involved the senile patients (over 65 years) with hip fractures to compare the expressions of peripheral and local IGF-I and IGFBP-3,4,5 and observe the correlation between expression and fracture type.Results The serum and local specimens of 46 patients were collected,with lower level of BMD,IGF-I and IGFBP-3,5 in the intertrochanteric fracture group than the femoral neck fracture group(P<0.001)and with higher level of serum and local IGFBP-4 in the intertrochanteric fracture group than the femoral neck fracture group (P<0.001).Only local IGFBP-4 mRNA had positive correlation with serum markers (R=0.544,P<0.001). Conclusions The expression difference of IGF-I and IGFBP in the peripheral and local serum may be one of pathogenesis leading to different types of hip fractures.Except for the IGFBP-4,the other factors may affect the bone metabolism of the senile hip fractures through different regulatory mechanisms.