Objective To explore effect of 5-HT,VEGF and mineral content in osteoporosis rat by Morinda officinalis.Methods 90 SPF male SD rats were selected, 15 rats were randomly selected as the normal control group, the rest were to establish the model of osteoporosis.When the model was established successfully according to the different treatment methods were divided into control group, model group, positive medicine group, low, middle and high does group.Bone mineral density, 5-HT, VEGF and the levels of the mineral were compared after 4 weeks treatment.Results Compared with the normal control group, BMD of model control group and low dose group was lower, Compared with the positive drug group, BMD of middle dose and high dose was higher ( P<0.05 ).Compared with model group,5-HT level of positive medicine group, high dose group and middle dose group was higher, compared with positive drug group, 5-HTP of middle dose group and high dose group was higher ( P <0.05 ).Compared with model group, positive drug group, VEGF levels of middle dose group and high dose group were higher(P <0.05), compared with positive drug group, VEGF of middle dose group and high dose group were higher(P<0.05), compared with model control group, positive drug group, serum P content of middle dose group was higher, compared with positive drug, serum P content of high dose group and high dose group was less.Conclusion Morindae Officinalis can increase 5-HT, VEGF in a rat model of osteoporosis, improve the level of serum P, has the guiding sense to the clinical.

Objective To discuss the influences of programmed death-1 (PD-1) factor on osteosarcoma cells MG-63.Methods Osteosarcoma stem cells were sovted and identified through osteosarcoma cell strain MG-63 cells.The influence of PD-1 signal on T cells proliferation were detected by MTT. The expression of PD-1 mRNA was detected by RT-PCR.Results Cancer cells had an obvious proliferation within one week, which showed the strong ability of proliferation and aggressivity.The formation of tumor cells spheres depended on the support of serum nutrition.The number of MG-63 cells proliferation in serum culture medium was significantly higher than the osteosarcoma cells spheres in serum-free suspension culture ( P<0.05 ) .Pluripotent stem cell marks that the expression of CD133 in cancer cell sphere was significantly higher than that of MG-63(P<0.05).RT-PCR results showed the PD-1 expression level of cancer cell sphere and MG-63 were increased significantly.Conclusion MG-63 cell line has the character istics of osteosarcoma stem cells.MG-63 cell line can express the corresponding cell markers.The expression of PD-1 also increase significantly which can reduce immune function of patients and is closely related with the occurrence and development of tumors.

At present, zebrafish has played an increasingly important role in models for human development and diseases and several areas of life sciences.As a newly laboratory animal resource, standardization research has become the technical bottleneck to be solved and an inevitable trend.In this review, we summarized the research history and character-istics of zebrafish and the status of quality standardization.We also discussed the main problem facing by the standardiza-tion research of zebrafish as a newly laboratory animal.We hope that the data can provide useful reference for the develop-ment of zebrafish quality standardization research.

Objective To establish a gata4 gene knockout zebrafish model of congenital heart disease, and construct transcription activator-like effector nuclease ( TALEN) vectors targeting gata4 gene.Method We construct TALEN vectors targeting zabrafish gata4 gene using unit assembly method and the in vitro-transcribed TALEN mRNAs were microinjected into one-cell stage zebrafish embryos.The efficiency of TALEN was identified by injected embryos, and mutations of zebrafish were screened and confirmed the different types through PCR and enzyme digestion.Results We successfully constructed correct targeting vectors by enzyme digestion and sequencing, and the gene knockout efficiency was 35.18%.We screened the mutant zebrafish and confirmed different types of gata4 gene mutations.Conclusions A gata4 knockout zebrafish model is successfully established, it can provide a good animal model for further research of congenital heart diseases.

Objective To analyze the cause and treatment of the postoperative complications using one-stage posterior spinal osteotomy in the treatment of severe spinal deformity.Methods From September 2006 to May 2013,17 patients with severe spinal deformity (congenital scoliosis in 11 cases,congenital kyphosis in 4 cases,and congenital kyphoscoliosis in 2 cases) underwent one-stage posterior spinal osteotomy,including 5 males and 12 females with an average age of 22.6 (14-51) years.The preoperative mean coronal Cobb angle was 109° (85°-160°) while the mean sagittal Cobb angle was 104° (65°-152°).Two patients had neurological symptoms preoperatively whose spinal cord function was D,evaluated by ASIA classification.All patients were treated with pedicle subtration osteotomy and pedicle screw internal fixation,which SPO osteotomy 2 cases,PSO osteotomy 11 cases,VCR osteotomy 4 cases.Results There were 17 cases of complications in 147 patients,the complication rate was 11.6％(17/147).The causes were as followed,screw malposition in 2 cases,compromised by close of resected areas in 2 case,residual bone compression in 1 case,acute spinal cord injury in 2 cases,infection in 2 cases,broken stick or loosen hat in 3 cases,and superior mesenteric artery syndrome in 5 cases.Postoperative neurological complications occurred in 7 cases.Two cases with preoperative ASIA D became ASIA C,5 cases with normal nerve function became ASIA C in 2 cases and ASIA D in 3 cases.After surgical exploration,given Methylprednisolone and neurotrophic drugs,removal or changing of the internal fixation,anti-infection and symptomatic treatment,15 cases recovered completely and 2 cases improved partially.Conclusion One-stage posterior spinal osteotomy for severe spinal deformity is technical demanding and risky,and the postoperative complications are common.Appropriate operative procedure,close observation of sensation and motor function,timely surgical exploration and nerve decompression,and early brace wear are all required.

Objective To assess the efficacy of one-stage posterior median incision via costotransverse joint in treating thoracic spinal tuberculosis.Methods Thirty three patients with tuberculosis of thoracic spine undergoing one-stage posterior thoracic spine debridement,bone grafting fusion and posterior instrumentation from July 2011 to October 2013 were included in the study.There were 18 males and 15 females.The age was from 17 to 39 years old with an average of 29.5.The course was from 5 to 11 months with an average of 7.3.The Cobb angle was from 19° to 42° with an average of 29.8°.There were 5 in upper thoracic spine,17 in middle thoracic spine and 11 in lower thoracic spine.6 were in Frankel scale grade C,11 were in grade D and 16 were in grade E before surgery.Postoperative kyphosis correction,recovery of neurological function and bone fusion were observed.Results All the surgeries were completed successfully.The operation time was from 120 to 230 min with an average of 183.The blood loss during operation was from 480 to 700 ml with an average of 530.All 33 patients were followed-up for 12-36 months with an average of 22.7 months.The Cobb angle was from 8° to 15° with an average of 10.2°,the correction rate was 77％.The Frankel scale of 11 patients recovered from D to E,2 recovered from C to D and 4 recovered from C to E.The postoperative kyphosis correction and Frankel scale were significantly improved,all patients had a 100％ bone fusion rate and there were no internal fixation loosened or shift of graft bone at the last follow-up.ESR and CRP were returned to the normal range and the tuberculosis symptoms disappeared.Conclusions One-stage posterior median incision via costotransverse joint can complete surgery by the same position and the same incision in treating thoracic spinal tuberculosis with safety and good clinical efficacy.

Objective To investigate and evaluate the changes of T lymphocytes and red cell immunity of peripheral blood in patients with primary hepatocyte carcinoma (PHCC) after radiofrequency ablation(RFA) treatment. Methods The pre- and post- RFA(3d,7d,14d) peripheral blood T lymphocyte subsets(T3,T4,T8,T4/T8) and red cell immunity (RBC C3 receptor flower and RBC-immuocomplex formation rate) were investigated in 120 patients with PHCC treated by RFA. Results On 7d, 14d after RFA, T3, T4 lymphocytes and T4/T8 were higher than those on preoperative day significantly(P

0.05). RFA showed a marked short term therapeutic effect for patients with PHC.Conclusions After RFA treatment of patients with PHC,the serum levels of TNF and AFP decreased,and this suggests an augmentation of organism immune function. The serum levels of TNF and AFP can assist in the assessment of the therapeutic effect and prognosis of primary hepatic carcinoma.

Objective:To investigate the difference of curative effect between zero-profile bridge-shaped locking cage (ROI-C) and anterior cage combined with titanium plate fixation in the treatment of two-level and three-level cervical spondylotic myelopathy.Methods:A total of 85 patients (43 males and 42 females), aged 52.3±8.0 years (range from 28 to 66 years) with bi- and three-level cervical spondylotic myelopathy who received surgical treatment from June 2017 to October 2019 were retrospectively analyzed. There were 63 cases of two levels and 22 cases of three levels. 45 cases were treated with zero-profile bridge-shaped locking cage ROI-C (ROI-C group), and 40 cases with anterior cage combined with titanium plate fixation (titanium plate group). The main observation indicators include operation time, intraoperative blood loss, cervical Cobb angle, fusion segment Cobb angle, average intervertebral height, pain visual analogue scale (VAS), Japanese Orthopaedic Association (JOA) Score and neck disability index (NDI).Results:All of 85 patients were followed up for 16.9±2.0 months (range 12 to 22 months). The operation time of two-level ROI-C group was 110.37±8.25 min, which was shorter than 139.5±10.54 min of titanium plate group; the intraoperative blood loss was 15.74±8.10 ml, which was less than 23.71±9.70 ml of titanium plate group; the operation time of three-level ROI-C group was 130.00±5.70 min, which was shorter than 162.83±5.59 min of titanium plate group, while the difference in the intraoperative blood loss between the two groups had no statistical significance. One year after operation, Cobb angle of cervical vertebra in double and three-level ROI-C groups were 15.31°±1.55° and 15.20°±0.42°, respectively, which were largerthan 11.23°±2.03° and 9.20°±1.14° before operation; in titanium plate group, they were 15.89°±1.13° and 16.08°±1.88°, which were higher than 11.25°±2.01° and 9.00°±1.60° before operation, and the differences had statistical significance. The differences between the two groups before operation and 1 year after operation had no statistical significance. One year after operation, the VAS scores of double and three-level ROI-C groups were 1.83±0.66 points and 2.60±0.52 points, respectively, which were less than the preoperative 7.49±0.51 points and 7.60±0.52 points; the titanium plate group was 1.79±0.50 points and 2.41±0.51 points, which were less than the preoperative 7.61±0.63 points and 7.42±0.52 points, and the differences had statistical significance. There was no significant difference between the two groups before operation and 1 year after operation. One year after operation, the JOA scores of double and three-level ROI-C groups were 15.00±0.84 points and 14.70±0.95 points, respectively, which were higher than the preoperative 7.20±0.87 points and 6.60±1.27 points; the scores of titanium plate group were 15.29±0.85 points and 14.83±0.58 points, which were higher than the preoperative 6.89±1.03 points and 6.92±0.67 points, and the differences had statistical significance. The differences between the two groups had no statistical significance. The postoperative JOA improvement rate was excellent. Postoperative dysphagia occurred in 1 case (2.22%, 1/45) in ROI-C group and 8 cases (20.00%, 8/40) in titanium plate group, and the difference in the incidence rate between two groups had statistical significance ( χ2=5.32, P=0.02). Conclusion:Both ROI-C and anterior cage combined with titanium plate fixation in the treatment of double and three-level cervical spondylotic myelopathy can achieve good short-term clinical efficacy, with shorter operation time and lower incidence rate of postoperative dysphagia using ROI-C.

Objective: To investigate the clinical effect of percutaneous curved vertebroplasty in the treatment of thoracolum-bar osteoporotic vertebral compression fractures (OVCFs). Methods: All of 85 patients with single thoracolumbar vertebral OVCFs who met the admission criteria from January 2017 to July 2018 were divided into three groups according to the random dig-its table method. They were treated with percutaneous curved vertebroplasty, routine unipedicular PVP and routine bipedicular PVP respectively. There were 25 patients in the percutaneous curved vertebroplasty group, 6 males and 19 females; aged 56-80 years, with an average age of 70.6±9.7 years. Fracture vertebral body distribution: T₁₀ 2 cases, T₁₁ 4 cases, T₁₂ 3 cases, L₁ 9 cases, L₂ 3 cases, L₃ 1 case, L₄ 1 case and L₅ 2 cases. There were 32 patients in the routine unipedicular PVP group, 6 males and 26 fe-males; aged 58-75 years, with an average age of 69.5±9.3 years. Fracture vertebral body distribution: T₁₀ 2 cases, T₁₁ 4 cases, T₁₂ 5 cases, L₁ 11 cases, L₂ 6 cases, L₃ 1 case, L₄ 1 case and L₅ 2 cases. There were 28 patients in the routine bipedicular PVP group, 5 males and 23 females; aged 59-81 years, with an average age of 69.8±8.8 years. Fracture vertebral body distribution: T₁₀ 2 cases, T₁₁ 4 cases, T₁₂ 4 cases, L₁ 10 cases, L₂ 4 cases, L₃ 1 case, L₄ 1 case and L₅ 2 cases. The operation time, injected cement volume, in-traoperative blood loss were recorded and analyzed. Preoperative, postoperative 1 week and 3 months visual analogue scale scores and oswestry disability index were adopted to value the clinical improvements. Preoperative, postoperative 1 week and 3 months relative vertebral height and kyphosis correction, and the cement leakage rate were measured and analyzed. Results: There was no significant difference in the data of gender, age, VAS scores, ODI and distribution of fracture vertebrae among the three groups (P>0.05), and the baseline data was comparable. The average VAS score in the percutaneous curved vertebroplasty group was 2.3±0.5 at 1 week after surgery, that of the routine unipedicular PVP group was 2.4±0.4 and that of the routine bipe-dicular PVP group was 2.4±0.4; the average ODI in the percutaneous curved vertebroplasty group was 19.8%±3.9%, that of the routine unipedicular PVP group was 20.0%±4.1% and that of the routine bipedicular PVP group was 19.9%±3.8%; they were lower than the preoperative data, which were statistically significant (P<0.001). The average relative vertebral height in the percutaneous curved vertebroplasty group was 48.99%±9.23% at 3 months after surgery, that of the routine unipedicular PVP group was 47.11%±10.12% and that of the routine bipedicular PVP group was 46.71%±11.16%; the average kyphosis cor-rection in the percutaneous curved vertebroplasty group was 6.21%±1.94%, that of the routine unipedicular PVP group was 5.22%±2.07% and that of the routine bipedicular PVP group was 5.97%±2.09%; there was 1 cement leakage case in the per-cutaneous curved vertebroplasty group; those of the routine unipedicular PVP group were 4 cases and those of the routine bipe-dicular PVP group were 6 cases; there was no significant difference among the three groups (P>0.05). Operation time 39.10±2.00 min vs 38.70±1.70 min, injected cement volume 3.60±0.11 ml vs 3.50±0.13 ml and blood loss 5.10±0.30 ml vs 5.00±0.40 ml of the percutaneous curved vertebroplasty group and the routine unipedicular PVP group were less than those of the routine bipedicular PVP group, which were statistically significant (P<0.05). Conclution Percutaneous curved vertebroplasty could achieve satisfactory clinical outcomes for OVCFs, with advantages of less operation time, less blood loss, limited X-ray expo-sure, less injected cement volume, and more balanced augmentation for stabilization of the affected vertebrae and total verte-bral column.