BACKGROUND: The meniscus has limited ability in repairing itself after being injured. However, tissue engineering provides a new way to meniscus repair after injury. Bone marrow mesenchymal stem cells (BMSCs), which possess the potential of multi-directional differentiations, can be ideal seed cells in meniscus tissue engineering. OBJECTIVE: To investigate the feasibility of differentiation of in vitro cultured porcine BMSCs into fibrochondrocyte phenotypes in inductive medium. METHODS: BMSCs were isolated with whole bone marrow culture method. Then, BMSCs of the third passage were digested and incubated in a medium containing transforming growth factor-β1, insulin-like growth factor-Ⅰ, dexamethasone and ascorbic acid in a 24-well plate at a density of 2.0×10~4/cm~2 in the experimental group. While in the control group, the DMEM-LG complete culture medium containing no inductive factor were used instead. At day 7, 14 and 21 after induction respectively, Toluidine blue staining and immunocytochemical staining were performed to detect differentiation. MAIN OUTCOME MEASURES: ①Population double time (PDT) of BMSCs; ②Morphological changes of BMSCs under light microscope;③Proteoglycan expression;④Collagen type Ⅰ and type Ⅱ expression. RESULTS AND CONCLUSION: ① The PDT of the second passage BMSCs was 2 days, which was the shortest. The PDT prolonged relatively after the fourth passage, which were 5 to 9 days. ② The BMSCs changed from a spindle-like appearance into a polygonal shape after induction. ③ In the experimental group, toluidine blue staining resulted in hyacinthine-stained cytoplasm and the blue was even deeper in the area where cells were dense; The degree of staining increased with the increasing induction time. While in the control group, only nucleus of BMSCs were stained blue. ④ Collagen type Ⅰ immunocytochemical staining was positive in both the experimental and the control group and there was no difference of significance between various induction time. No collagen type Ⅱwas seen expressed in the control group, while in the experimental group it was seen to be expressed steadily after 14 days of induction. It is indicated tlat BMSCs can be induced to synthesize fibrochondrocyte-characterized extracellular matrixes in vitro, which suggests that BMSCs are available as seed cells in meniscus tissue engineering.

Objective To determine the outcome of the combined use of flaps transfer and ilizarov technique reconstruct the large soft tissue defects and bone lose in the lower leg.Methods Sixteen patients were identified from a retrospective review from July,2008 to July,2013,who suffered the large soft tissue defects and bone lose in the lower leg and underwent single-stage soft tissue and osseous reconstruction using the flap technique and Ilizarov method.There were 12 males and 4 females aged from 22 to 62 years old (average 42.6 years old).The size of soft tissue defect ranged from 8 cm×9 cm to 30 cm×20 cm.The length of the bone discrepancy ranged from 2 to 14 cm.According to the local condition of the lower leg and the size of the composite tissue defects,10 patients received the free flap covering,6 patients repaired by the saphenous neurocutaneous perforator flap (3 cases) and sural nerve neurocutaneous flap (3 cases).Daily monitoring the skin temperature postoperative.Distraction was commenced on postoperative day 10 to 14 at the rate of 1 mm/day and continued in 4 equal increments.Results The follow-up time ranged from 18 to 36 months.Sixteen flaps survived completed,only 1 flap was observed the venous congestion in postoperative day 2.The duration of ilizarov application ranged from 3.5 to 18.0 months.All patients achieved final union.All patients were satisfied with the outcome of the surgery.Conclusion The combined use of neurocutaneous flap and Ilizarov technique for reconstruction of large composite soft tissue defect in the lower leg.Significantly reduce patient treatment time,improving traction osteogenesis of long bones and the ability of resistance to infection.

ObjectiveTo study the fluoride level of preschool children influenced by fluoride toothpaste. MethodsAbout 200 children aged 3～6 years old were involved in the test. The children of test group were given fluoride toothpaste while the children of control group were given non-fluoride toothpaste. After two months, hair samples were gasified by high temperature and were tested with fluoride-specific electrode. Urinary fluoride concentration and 24h urinary fluoride excretion were also conducted by using fluoride-specific electrode.ResultsIt shows that the 24h urinary fluoride excretion and the fluoride concentration in hair and urine of children in control group were apparently lower than those of children in test group.ConclusionFluoride toothpastes used by preschool children may result in the change of fluoride level in their bodies under the stability of their daily habits and living cir cumstances. Age is likely to be one of the facts that can influence the fluoride ingestion level.

Objective The incidence rate of complication during central venous catheterization through internal jugular vein (IJV) and subclavian vein (SV) puncture. Methods Clinical controlled trials about IJV and SV puncture were collected and related literatures were screened according to the criteria of inclusion. The literatures underwent Meta analysis and subsequent analysis of sensitivity. Results A total of 18 literatures were included. Meta analysis indicated that statistical difference existed in the related infection rate [RR=1.74, 95%CI (1.32, 2.30)] and arterial puncture [RR=3.19, 95%CI (1.70, 5.99)], but not in the one-time-puncture success rate [RR=1.06, 95%CI (0.90, 1.24)] between IJV and SV puncture. Conclusions The rate of related infection and arterial puncture was higher by IJV than by SV puncture. But we could not confirm if any difference existed in the one-time-puncture success rate between the two methods. The results still needs evaluation by high-quality randomly controlled experiments.

Objective To detect the preliminary results of combined periacetabular and intertrochanteric osteotomies for complex hip dysplasia.Methods From January 2006 to August 2011,23 patients (25 hips) with complex hip dysplasia were treated with combined periacetabular and intertrochanteric osteotomies (ITO).Valgus ITO was performed on 2 hips and varus ITO on others.The averagc age of patients at the time of index procedure was 20.5± 3.9 years (range:15 to 26).The hip pain period before surgery varied from 5 to 24 months (mean 11±4.7 months).The Shenton's line was broken in every hip and the mean preoperative lateral central edge angle (LCEA) was 5.6°±4.6°.The average preoperative fenoral neck-shaft (CCD) angles were 158°±3.2° and 110° in 23 valgus hips and both of two varus hips respectively.The preoperative Harris hip score was 76.7±3.7.According to T(o)nnis osteoarthritis grades,there were 7 hips at grade 0,12 hips at grade Ⅰ and 6 hips at grade Ⅱ.Results The mean followup period was 40±18 months (range:12 to 78).The CCD angle (130°±2.1°),LCEA (28.6°±2.9°) and Harris hip score (90.8±3.3)were all improved postoperatively,whereas no progression was found in preoperative T(o)nnis grades.The postoperative Harris hip scores were excellent in 17 hips and good in 8 hips.The T(o)nnis osteoarthritis grades were grade 0 in 6 hips,grade Ⅰ in 11 hips and grade Ⅱ in 8 hips.There wcre six cases with lateral femoral cutaneous nerve injury,three cases with limp and two cases with broken Shenton's line.Neither femoral head necrosis nor bone nonunion was found in this study.Conclusion Satisfactory preliminary results could be obtained through combined periacetabular and intertrochanteric osteotomies among patients with complex hip dysplasia.The risk of femoral head necrosis and bone nonunion was low.

Objective To detect the expression of peripheral blood free prostate specific antigen (fSPA),total prostate specific antigen (tSPA),prostate specific antigen density (PSAD) and prostate cancer gene 3 (PCA3) in prostate disease,and the significance of combined detection of fSPA,tSPA and PCA3.Methods 67 patients with prostate cancer,75 patients with prostatic hyperplasia and 70 healthy male were selected as the research objects from Dec.2014 to Jul.2016.The serum level of fSPA and tSPA was detected by chemiluminescence immune staining method.The prostate volume was tested by ultrasonic sound and PSAD value was calculated.The total RNA was extracted by Trizol,and the serum PCA3 mRNA expression was detected by RT-PCR.The specificity and sensitivity of combined detection of fSPA,tSPA and PCA3 were analyzed.Results The serum levels of fSPA,tSPA,PSAD and PCA3 in prostate cancer were significantly higher than those in patients with prostatic hyperplasia and healthy male,and they were higher in patients with hyperplasia of prostate than in healthy male,and the differences had statistical significance (P＜0.01).The serum levels of fSPA,tSPA,PSAD and PCA3 were higher in patients with Gleason score ≥7 points and in T3-T4 stage than in patients with Gleason score ＜7 and in T1-T2 stage,and the difference had statistical significance (P＜0.01).The serum levels of fSPA,tSPA,PSAD and PCA3 were positively correlated with Gleason score and TMN pathological stage,and the difference had statistical significance (P＜0.01).The AUC value of fSPA,tSPA,PSAD and PCA3 in diagnosis of prostate cancer was 0.53,0.57,0.63 and 0.75,and the AUC value of combined detection was 0.92.The combined detection efficiency was higher than the single index.The specificity of fSPA,tSPA,PSAD and PCA3 was 67.16％,68.66％,73.13％ and 85.07％,and the sensitivity was 71.64％,70.15％,74.63％ and 82.09％ respectively.The specificity of combined detection was 97.01％,the sensitivity was 92.54％,and the difference had statistical significance (P＜0.01).Conclusion The serum level of fSPA,tSPA,PSAD and PCA3 is increased in prostate disease,and is negatively correlated with Gleason score and TMN pathological stage.The combined detection of fSPA,tSPA,PSAD and PCA3 can improve the sensitivity and specificity of prostate disease diagnosis,and is of high clinical value.

Objective To evaluate the effect of rehabilitation training on dysphagia and trismus in patients with nasopharyngeal carcinoma after radiotherapy.Methods Fony-three post-radiotherapy nasopharyngeal carcino-ma patients were divided into a rehabilitation group and a control group.Both groups were subjected to routine treat-ment,while the rehabilitation group received rehabilitation training in addition.The patients were assessed with a wa-ter-swallowing test of swallowing.Late effects of normal tissues/subjective and objective medical analysis(LENT/SOMA)scored and inter-incisor distance were measured to assess trismus before and after treatment.Results The rehabilitation group displayed significant improvement in swallowing as well as increased inter-incisor distance.Con-clusions Rehabilitation training can improve swallowing,prevent or delay trismus and improve the quality of life of patients.

Objective To report the operative technique and clinical results of reversed perforator-plus sural neurofasciocutaneous flap in lower leg. Methods Perforator-plus sural neurofasciocutaneous flaps, which designed along the axis of the sural nerve and based on the dual-pedicle of fasciocutaneous and per-oneal perforator, were used to reconstruct soft-tissue defect in lower extremities. Results Twenfy-four flaps, ranged from 25 cm×12 cm - 8 cm ×7 cm in size, survived completely without venous congestion and distal ischemia and necrosis. The color and texture of the flaps were good. The appearance and functional re-suits were satisfactory with following up for 6 to 12 months. Conclusion The modified technique in pediclehave minimized the complication, enlarged the size of the flap and improved the survive rates. It is a goodmethod in repairing large soft-tissue defects of extremities.

AIM:To observe the expression of tyrosine hydroxylase (TH), ionized calcium binding adapter molecule 1 (Iba1) and pro-opiomelanocortin (POMC) in rat arcuate nucleus (Arc) of hypothalamus induced by substantia nigra (SN) lesion with 6-hydroxydopamine (6-OHDA).METHODS:According to the random number table, 20 SD rats were randomly divided into 6-OHDA group and control group.6-OHDA was injected into the bilateral SN of the rats in 6-OHDA group, and the same volume of saline was injected into the same position of the rats in control group.After 6 weeks, the rats were sacrificed and the brains were obtained.Immunofluorescence staining and Western blot were used to detect TH expression in the SN, and TH, Iba1 and POMC expression in the Arc.RESULTS:Compared with control group, no significant change of body weight in 6-OHDA group was observed, but the weight of retroperitoneal fat decreased from (7.550±0.670) g to (3.895±0.465) g (P<0.01).The number of TH immunoreactive neurons in SN decreased from 56±5 to 14±2 (P<0.05), and the TH protein level decreased from 0.75±0.11 to 0.41±0.09 (P<0.01).In the Arc, the number of TH immunoreactive neurons decreased from 21±2 to 10±3 (P<0.05), the number of Iba1 immunoreactive neurons increased from 12±2 to 30±5 (P<0.05), and the number of POMC immunoreactive neurons increased from 16±4 to 31±2 (P<0.05).In the hypothalamus, the TH protein level decreased from 0.35±0.05 to 0.21±0.02 (P<0.01), the Iba1 protein level increased from 0.23±0.06 to 0.51±0.04 (P<0.01), and the POMC protein level increased from 0.37±0.05 to 0.65±0.03 (P<0.01).CONCLUSION:The changes of TH, Iba1 and POMC expression in Arc of 6-OHDA-treated rats may involved in the fat loss in Parkinson's disease.