BACKGROUND: The establishment of animal models is of important significance for understanding the pathogenesis, prevention, and treatment of intervertebral disc degeneration. OBJECTIVE: To develop rabbit models of intervertebral disc degeneration by acupuncture and to analyze the effects of acupuncture on intervertebral disc height and degeneration by X-ray and MRI examinations. DESIGN: A randomized and controlled observation. SETTING: Department of Orthopaedics, Changhai Hospital, Second Military Medical University of Chinese PLA. MATERIALS: This study was performed at the Animal Cener, Changhai Hospital, Second Military Medical University of Chonese PLA between June 2005 and April 2006. Six healthy New Zealand white rabbits of either gender, averaging 6 months old, weighing 2.5 kg on average, were provided by Animal Cener, Changhai Hospital, Second Military Medical University of Chinese PLA [permission No. SCXK (hu) 2002-0006], and recruited for this study. The protocol was performed in accordance with ethical guidelines for the use and care of animals.METHODS: ① Operation was made in experimental rabbits via the anterior retroperitoneal approach. The management for the intervertebral discs was as follows: L3-4 was the control disc without exposure or needle stab; L4-5 was subjected to sham-operation with only exposure; L5-6 was the experimental disc with three times of anterior-lateral needle stab after exposure. Lumbar lateral X-ray film was respectively taken with a CR machine (Simens Company) prior to and 4 weeks after surgery for measuring L3-4, L4-5, and L5-6 intervertebral disc height and calculating the ratio with intervertebral disc height prior to surgery. Each segmental lumbar intervertebral disc T2-weighted signal was measured with a 1.5 T medical superconductive magnetic resonance scanner (Simens Avanto). According to signal intensity, intervertebral disc degeneration was graded: a score of 4 indicated normal intervertebral disc without apparent degeneration but with bright signal, a score of 3 indicated slight degeneration with a little decrease in T2-weighted signal, a score of 2 indicated moderate degeneration with apparent decrease in T2-weighted signal, and a score of 1 indicated severe degeneration with apparent decrease in both T2-weighted signal and intervertebral disc height. MAIN OUTCOME MEASURES: Intervertebral disc height change revealed by lumbar lateral X-ray film; Intervertebral disc degeneration degree analyzed through magnetic resonance imaging (MRI) examination. RESULTS: All 6 experimental rabbits were included in the final analysis. The postsurgery-to-presurgery ratio of intervertebral disc height was 0.982 5±0.017 08 and 0.972 5±0.017 08 for L3-4 and L4-5 respectively, which was higher compared to that for L5-6 (0.550 0±0.02 582). Significant statistical difference existed (P ＜ 0.01). T2-weighted singal score was (4.00±0.0) and (3.75±0.5) for L3-4 and L4-5 intervertebral disc, respectively, which was higher compared to that for L5-6 intervertebral disc. Significant statistical difference existed (P ＜ 0.01). CONCLUSION: After 4 weeks of acupuncture, intervertebral disc height is decreased and interterbral disc degeneration obviously appears.

AIM:In human being,the exposure of lumbar Intervertebral discs usually requires the anterior transperitoneal or lateral retroperitoneal approaches. To study the feasibility of retropedtoneal approach to the lumbar spine in the New Zealand white rabbits.METHODS: The experiment was carried out between April 2005 and April 2006 in the Animal Center of Changhai Hospital.Second Military Medical University of Chinese PLA.Thirty New Zealand white rabbits,aged 6 months old, of either gender, weighing 2.5 kg,provided by the Animal Center of Changhai Hospital,Second Military Medical University of Chinese PLA [permission No.SCXK (hu) 2002-0006],were used in this study.During the experiment,the management of animals corresponded to Animal Ethical Standard.This study was approved by the Experimental Animal Ethics Committee,Changhai Hospital,Second Military Medical University of Chinese PLA.Transperitoneal approach was used to exposure the lumbar spine of experimental rabbits. The skin was disassociated, and a blunt dissection was made backwardly along the juncture of the maximum back muscle and external oblique muscle of abdomen,and lumbar intervertebral disc was exposed gradually.Observation index:①Operative effect of exposure.②Postoperative general condition (food intake,urination and defecation,activity,incision healing and death or not).RESULTS: Thirty experimental rabbits were involved in the final analysis.①The operative effect of exposure:The dght retroperitoneal approach was chosen,with satisfactory anesthesia and excellent exposure of rabbit lateral lumbar spine. And the exposure was usually accomplished within 15 minutes, with Less bleeding and no retroperitoneal rupture. ②Postoperative general condition: All animals recovered their consciousnesses within about 1 hour. They were intravenously injected continuously for supplementing blood volume,and after that,they were raised in the separate cases and were allowed to move freeLy.On the day of operation.they were fasted and forbidden to access to water.Their appetites began to decrease on the postoperative 1 day.restored from the 3rd day postoperatively and restored to normal on the 40th day postoperatively.All animals survived until after 4 weeks.They could move freely,without paralysis occurred or incontinence. There was one experimental animal whose surgical Incision broke.Debridement and resuture were given, and its wound healed. In all the other animals, the surgical incision healed well.CONCLUSION: Retroperitoneal approach can give an excellent access to the lumbar spine with easy operation and less complications.

Objective To evaluate the uncertainty for the determination of salicylic acid in human plasma by ultra performance liquid chromatography(UPLC).Methods All sources of uncertainty in the whole process of salicylic acid determination were analyzed,then the combined and expanded uncertainty were evaluated.Results The expanded uncertainty at concentrations of the lowest limit of quantitation(0.23 μg·mL-1) and a high level(39.43 μg·mL-1) of salicylic acid(P=95%,k=2) was 0.014 and 7.34 μg·mL-1,respectively.Conclusion The uncertainty of salicylic acid determination in human plasma by UPLC was mainly caused by recovery,repeatability and sample preparation at the lowest limit of quantitation and high qulity control concentration.

Objective To develop an in vitro model for cranial suture of neonatal SD rat. Methods The parietal hones with the sagittal suture were removed from SD rats (19-days old) for organ culture. In the experimental group, tensile force 3.92×10~(-3) N (0.4 g) was applied by helical springs, whereas no tension (0 N) was set in control group. These explants were observed under inverted microscope. At the end of the incubation period for 24 hours, general conditions were observed under inverted microscope and histological conditions were observed after hematoxylin and eosin stain. Results Under inverted micro-scope, sutures had no obvious changes in control group, whereas sutures were enlarged gradually in the experimental group. With histological observation, sutures developed normally in control group, but in experimental group, osteohlasts and capillary vessels proliferated actively in the suture. Conclusions In vitro model of cranial suture can be cultured and grown successively.

BACKGROUND: Previous studies demonstrated that recombinant human bone morphogenetic pretein-7 (hBMP-7) can effectively promote the synthesis of extracellular matrix, repair of damaged disc matrix, restore of degenerative disc height. It is hoped that BMP-7 can be used to control and reverse the intervertebral disc degeneration. However, because of the short half-life of recombinant protein and low biological activity, it is difficult to maintain BMP-7 high concentrations on degenerative disc. Gene therapy can prevent these defects effectively.OBJECTIVE: To study the effect of hBMP-7 gene transfeotion on biological activity of primary cultured rabbit nucleus pulposus cells in vitro, to determine the feasibility of hBMP-7 gene which will on gene therapy research of intervertebral disc degeneration,and to provide basis for further study.DESIGN, TIME AND SETTING: A randomized and controlled observation was performed at the Institute of Cardiothoracic Surgery of Changhai Hospital from December 2005 to September 2006.MATERIALS: Six healthy New Zealand white rabbits of either gender, averaging 4 weeks old and weighing 500 g, were used in this study, and Ad-hBMP7 was constructed by the Cardiothoracic Surgery Institute of Changhai Hospital.METHODS: After rabbits sacrifice under aseptic condition, the nucleus pulposus was got. After digested with Pronase, type Ⅱcollagenase and type Ⅱ DNAase (4 hours, 37 "(2), the cells were seeded in the Petri dishes and were kept in the incubator. After 7 days and then twice a week, the media were refreshed. The nucleus pulposus cells were infected by adenovirus integrated with hBMP7 gene. The cells which were transfected by adenovirus vector for Lac-Z gene and which were not transfected were adopted as control.MAIN OUTCOME MEASURES: The expression of hBMP-7 was determined by RTopCR and Western blot. The effect of hBMP-7 on cell proliferation was surveyed by MTT. Furthermore, the effect of hBMP-7 gene on synthesis of proteoglycan and type Ⅱcollagen was detected by modified dimethylmethylene blue method and ELISA, respectively.RESULTS: Gene sequencing and PCR showed that hBMP-7 gene was inserted correctly and no mutation happened. The pdmary cultured rabbit nucleus pulposus cells were identical with those reported in literature. After Ad-hBMP7 transduct into the nucleus pulposus cells, high level of hBMP-7 expression was observed and lasted over 3 weeks. Also hBMP-7 gene can promote cell proliferation and synthesis of proteoglycan and type II collagen with significant difference compared with control groups (P < 0.05).CONCLUSION: hBMP-7 gene mediated by adenovirus can be the target gene for the further study on gene therapy of intervertebral disc degeneration.

Objective According to the fine artery anatomy of the lower lip, we developed an innovative partial-thickness myocutaneous flap based on the vascular network of the submucosal and subcutaneous layers of the lower lip.We attempted to treat the secondary bilateral cleft lip deformities using this innovative cross-lip flap.Methods From July 2009 to June 2015, this new technique was used in 98 patients with secondary bilateral cleft lip deformities.The central defects usually occurred in these patients.The defects were reconstructed partially or completely according to the severities of the defects.The operative procedures were as follows: The split flap was elevated from the posterior portion of the oris orbicularis muscle after the inferior labial arteries were divided.Then the partial-thickness flap was rotated 180 degrees horizontally and inverted 180 degrees upward to the upper lip defect.Results All 98 musculomucosal pedicle flaps were viable.The upper lips were reconstructed according to the severities.Conclusions This myocutaneous cross-lip flap with musculomucosal-pedicle has a more reliable blood supply, better flexibility and plasticity.It is an effective method for aesthetic reconstruction for the secondary bilateral cleft lip deformities.

Objective: To evaluate the efficacy of the third-generation instrumentation including TSRH, CD and ISOLA in the treatment of adult scoliosis. Methods:Thirty-five adult patients with idiopathic or degenerative scoliosis who received treatment with third-generation instrumentation (TSRH,CD and ISOLA) between July 1999 to January 2003 were retrospectively reviewed. The mean preoperative cobb angle of major curves of the frontal plane was 58.1°(42°-95°). The patients received a combined anteroposterior approach or a single posterior procedure. The mean follow-up time was 20 months(10-48 months). Preoperative and postoperative Cobb angles of the frontal plane and sagittal plane and the distance between C7 and CVLS were measured. The subjective assessment was judged by questionnaire. Results: Postoperative clinical appearance of all patients improved significantly. Mean correction of major curves of the coronal plane was 53.2%. Mean loss of correction of the coronal plane in the last follow-up was 4.3°. The distance between the midline of C7 and CVSL was corrected from 2.6 cm to 0.24 cm. The results of follow-up showed that 89.3% patients were satisfied with the outcome. Pneumatothorax and haematothorax occurred in 2 patients. Three patients still complained of low back pain one year after operation because of adjacent degeneration in 2 patients and pseudoarthrosis in the remaining 1 patient. Conclusion: Imageologic findings and subjective assessment of the patients showed that the third-generation instrumentation can achieve good correction and trunk balance in the treatment of adult scoliosis with fewer complications.

Objective To explore the efficacy of multiple interventional models combined through anterior tibial veins approach in treatment of acute deep venous thrombosis (DVT).Methods The clinical data and the imaging materials of 10 patients with acute DVT were analyzed retrospectively,and the interventional treatment strategies were discussed.Results The percutaneous mechanical thrombectomy (PMT) was performed after a temporary inferior vena cava filter implanted in all patients,and thrombus clearance rate of grade Ⅲ was obtained in 7 patients while 3 patients reached grade Ⅱ criteria.There were 7 patients underwent angioplasty,then the inferior vena cava filter was retrieved during the same procedure in 5 patients while 4 patients in the second stage and 1 patient rejected to retrieve the filter.All swelling limbs alleviated significantly after those procedures.There was no critical complications occured.The total hospital stay ranged from 4 to 7 days,with a mean time of (6.00±0.94) days.All patients were followed up with a mean time of (3.60± 1.84) months (ranged from 1 to 6 months).No thrombus recurrences of the previously affected vessels were reported.Conclusion The multiple interventional models combined via anterior tibial veins for acute DVT is simple,safe and effective,combined with mechanical thrombectomy device can reduce thrombus burden quickly,and combined with angioplasty can consolidate the therapeutic effect,all those methods combined can shorten the overall hospitalization time.

This study is to report the establishment of an UPLC-MS/MS method for the determination of plasma concentration of UA carried in self-microemulsifying drug delivery system (SMEDDS) and its pharmacokinetics in rats. It was used for determination and analysis when serum with internal standard was extracted from C18 solid-phase column. Acquity UPLC BEH C18 column (100 mm x 2.1 mm, 1.7 microm) was used for separation. The mobile phase was acetonitrile -0.1% ammonia with gradient elution at the flow rate of 0.2 mL x min(-1). The column temperature was 40 degrees C and the detection wave length was 210 nm. It was detected by negative ion using electrospray ionization source (ESI) and scanned by multiple reaction ion monitoring (MRM) mode. The liner relationship of UA was very good in the range of 1.19-3 815.00 ng x mL(-1) (r = 0.999 0). Recovery rate of different concentrations were 87.42%-89.95%. The precision of inter-day and intra-day were less than 11%. The method developed in our study was proved to be sensitive, rapid and simple. It is suitable for the pharmacokinetic study of UA-SMEDDS in rats.

Objective To investigate the changes of CD4+ CD25+ regulatory T lymphocyte (Treg) and expressions of folkhead helix transcription factor 3 (FoxP3) in intestinal mucosa in human immunodeficiency virus (HIV) infected patients. Methods Twenty-one HIV infected patients and 17 control subjects without HIV infection were included in this study. The expression of FoxP3, which was considered as a specific marker of CD4+ CD25 + Treg, was detected in intestinal mucosa specimens from HIV infected patients by immunohistochemistry. Meanwhile, the in situ expression of CD4+ T lymphocyte was also determined by immunohistochemistry. The data were analyzed by t test. Results The positive labeling index of CD4+ T lymphocyte in intestinal mucosa was significantly lower in HIV infected patients compared to the controls (11. 56%±4. 44% vs 43. 49% ±8. 90% ,t=-11. 86,P<0. 01). The positive labeling index of FoxP3 in intestinal mucosa was also significantly lower in HIV infected patients compared to the controls (0.46% ± 0.20% vs 1. 18% ± 0. 44% ,t= - 5. 98,P<0.01). Conclusion The depletion of CD4+ CD25+ Treg is accompanied with the depletion of CD4 + T lymphocyte and the reduction of FoxP3 expression in intestinal mucosa of HIV infected patients.