BACKGROUND:Intervertebral disc can bear load but lack vessels. Nucleus pulposus cel s have the problem of phenotype loss during in vitro culture that can lead to degenerative changes. The mechanism of intervertebral disc degeneration remains unclear.
OBJECTIVE:To explore the approaches of isolation, adherence culture, amplification and identification of the rabbit nucleus pulposus cel s in vitro, to observe the growth characteristics of nucleus pulposus cel s in different generations.
METHODS:Type Ⅱ col agenase digestion method plus explants culture method was used to isolate and purify nucleus pulposus cel s and then amplify in vitro. The morphology and growth of primary and passaged cel s was observed under the inverted microscope, the number of cel s was counted and the growth curve was draw. The morphology of the cel s was observed under light microscope after hematoxylin-eosin staining, and the expressions of col agen type Ⅱ and aggrecan were examined by immunocytochemistry.
RESULTS AND CONCLUSION:Nucleus pulposus cel s of rabbit were isolated, cultured and amplified in vitro successful y. Growth activity was observed, and found that the 1-3 generation nucleus pulposus cel s proliferated more rapidly and vigorously. The proliferation of nucleus pulposus cel s was decreased while the cel passaged more generations. These isolated and cultured nucleus pulposus cel s could positively express the col agen type Ⅱ and aggrecan. In vitro combination of type Ⅱ col agenase digestion method and explants culture method could obtain high purity nucleus pulposus cel s, and the cultured nucleus pulposus cel s were grew in round or polygonal. The 1-3 generation of cel s had strong activity.

Objective The clinical features of elderly patients with severe acute pancreatitis (SAP) were atypical and these patients were often misdiagnosed as having paralytic ileus. The clinical presentations of elderly patients with SAP whose first diagnosis as paralytic ileus were analyzed. Methods 18 patients of elderly SAP who were misdiagnosed as having paralytic ileus were included and the clinical data were compared with 58 elderly patients with SAP. Results Among the misdiagnosis group, the first symptom onset were fleus, abdominal distension, vomiting, constipation, abdominal pain, diarrhea for 5, 4, 3, 3, 2, 1 case, respectively. Among SAP group, the first symptoms onset were 2, 31, 9, 3, 11, 2 cases, respectively. For misdiagnosis group, 13 cases were correctly diagnosed by CT scan, 3 cases by ultrasound and 2 cases by serum amylase test. For SAP group, 32, 15, 11 cases were diagnosed by CT scan, ultrasound and serum amylase, respectively (P < 0.05). 4 and 13 patients died in misdiagnosis and SAP group, respectively; among these 13 patients, 10 were female and 3 were male. Conclusions The elderly patients with paralytic ileus should consider the possibility of SAP, and CT scan was valuable for correct diagnosis.

BACKGROUND:Percutaneous kyphoplasty for osteoporotic vertebral fractures with posterior vertebral wal breakage can relieve pain rapidly, but there is a risk of leakage of bone cement. OBJECTIVE: To observe the effect of bone filing mesh container in percutaneous kyphoplasty for osteoporotic vertebral fractures with posterior vertebral wal breakage. METHODS: Forty senile patients with osteoporotic vertebral compression fractures were enroled, whose CT and MRI showed different degrees of posterior vertebral wal breakage, including 19 males (21 vertebral bodies) and 21 females (28 vertebral bodies), aged 50-87 years. These 40 patients were subjected to percutaneous kyphoplasty, and bone filing mesh container was used to deliver bone cement. Then, changes in visual analogue scale score, vertebral height and leakage of bone cement were observed in patients before and after treatment. RESULTS AND CONCLUSION:The surgery was successful in al the 40 patients, and no pulmonary embolism, cement leakage, and spinal cord and nerve root injuries appeared. Al the patients were folowed for 10-12 months. The visual analogue scale scores and vertebral height were improved significantly at both 1 week and 3 months after treatment compared with those before treatment, but there was no difference in the visual analogue scale scores and vertebral height at 1 week and 3 months after treatment. These findings indicate that percutaneous kyphoplasty with bone filing mesh container is effective to prevent bone cement leakage.

Objective To explore the function and clinical effect of automatic nerve retractor in micro-endoscopic discectomy.Methods From August 2009 to December 2010,350 patients with lumbar disc hemiation were treated by micro-endoscopic discectomy,including 196 males and 154 females,aged from 17 to 68 years (average,42 years).Three cases were at L1-2,8 at L2-3,12 at L3-4,186 at L4-5 and 141 at L5S1.The automatic nerve retractor was used in all micro-endoscopic discectomy.The visual analogue scale (VAS) and Oswestry disability index (ODI) were used to evaluate clinical outcomes.Results All patients were followed up for 6 to 16 months (average,9 months).The mean VAS score decreased from preoperative 8.79±1.15 to 3.80±1.14 3 months after operation and 3.65±1.14 6 months after operation.The mean ODI score decreased from preoperative 78％±1.71％ to 28％±1.72％ 3 months after operation and 28％±1.88％ 6 months after operation.Postoperative VAS and ODI scores decreased significantly compared with those before operation.The VAS and ODI scores 6 months after operation were not significantly improved compared with those 3 months after operation.No spinal cord and nerve root injury and epidural hematoma formation occurred in all cases.Conclusion In micro-endoscopic discectomy,the automatic nerve retractor can help the operator obtain effective exposure,protect nerves from injury,alleviate workload of the operators,therefore it has a great clinical application value.

Objective To explore the clinical the efficacy of large diameter esophagogastric internal fistula combined with fundoplication and modified Heller surgery for treatment of achalasia.Methods From June 2008 to March 2014,18 patients diagnosed as achalasia were se-lected into this suty,and they were divided into the observation group (8 cases)and the control group (10 cases).Patients of the observation group were received large diameter esophagogastric internal fistula combined with fundoplication while patients of the control group received modified Heller surgery.Compared the surgical curative effect and complications between the two groups.Results All the 18 patients suc-cessfully completed the arranged surgical treatment.The imaging examination 3 months after the operation showed that the contrast agent suc-cessfully passed through the esophagus,cardia and the anastomosis,and then went into the gastric lumen.The total effective rate was 100% in the observation group 12 months after operation,while it was 60% in the control group,and the difference between the two groups was statisti-cally significant (P =0.014).The incidence of complications in the observation group was lower than that of the control group,the difference was statistically significant (P =0.026).Conclusion Large diameter esophagogastric internal fistula combined with fundoplication for treat-ment of achalasia is safer,and there is a possibiltiy to cure the achalasia.

BACKGROUND:Transplantation of mesenchymal stem cels to prevent and treat degeneration of the intervertebral disc is a feasible method. Mesenchymal stem cels co-transfected by SRY-related high mobility group-box gene 9 (SOX-9) and growth differentiation factor-5 (GDF-5) can differentiate into nucleus pulposus cels, in order to obtain greater effect of induction and proliferation of nucleus pulposus cels. OBJECTIVE:To investigate the effect of SOX-9 and GDF-5 co-transfection on the differentiation of rabbit bone marrow mesenchymal stem cels into nucleus pulposus cels. METHODS: We separated and cultured bone marrow mesenchymal stem cels from the bone marrow of rabbit aged 4 months. Passage 3 cels were divided into five groups andin vitro induced to differentiate into nucleus pulposus cels: non-transfected group, empty vector transfection group, SOX-9 transfection group, GDF-5 transfection group, SOX-9 and GDF-5 co-transfection group. At 14 days after transfection, RT-PCR was employed to assay SOX-9, GDF-5 and colagen type II mRNA expressions in bone marrow mesenchymal stem cels. The marker of nucleus pulposus cels-KRT19 expression was also detected by immunohistochemical staining. RESULTS AND CONCLUSION:In the co-transfection group, the mRNA expressions of SOX-9, GDF-5, and colagen type II were significantly higher than those in the SOX-9 transfection group, GDF-5 transfection group, and both these two groups, respectively (P < 0.05). Cels were positive for KRT19 in the SOX-9 and GDF-5 groups, and strongly positive for KRT19 in the co-transfection group. These findings indicate that double gene-transfected bone marrow mesenchymal stem cels are better than single gene-transfected cels with regard to differentiation into nucleus pulposus cels and secretion of extracelular matrix.

Objective To clone ,construction ,express and purify Tp47 of Treponema pallidum (Tp) ,and assess the immunoreac‐tivity by Western‐Blot .Methods Tp47 was amplified by polymerase chain reaction ,and then cloned to the vector pGEX‐6P‐1 .The correct sequence of the recombinant plasmids pGEX‐6P1‐Tp47 was transformed into Escherichia coli BL21 (DE3) and induced .The expression product was analyzed by sodium dodecyl sulfate polyacrylamide gel electropheresis and Western‐Blot .The expression protein was purified .Serum of different clinical stages of syphilis was used as the antibody to detect the immunoreactivity of the protein by Western‐Blot .Results A fusion protein with molecular weight about 71 × 103 was attained .Western‐Blot proved that the recombinant protein can react with Tp IgG positive sera .And the specificities and sensitivities of the diagnostic reagent detected by sera were 100% .Conclusion The recombinant protein Tp47 was expressed and purified with good antigen activity ,which could provide the basis of theory and practice for the development of early diagnostic kit applying to detect Tp infection .

Objective:To investigate the inhibitory effect of the water-soluble preparation of curcumin on tumor angiogenesis induced by alginate-encapsulated colon carcinoma C26 cell line in mice. Methods: The water-soluble preparation of curcumin was made by forming inclusion compound of curcumin with hydroxypropyl-?-cyclodextrin (C-HP-?-CD). Subcutaneous imbedding of alginate-encapsulated colon carcinoma C26 cells was used to create tumor bearing model in BALB/c mice. Then the mouse models were intraperitoneally injected with C-HP-?-CD, TNF-70 (as positive control), or HP-?-CD(as negative control). The angiogenesis in the alginate beads was observed under light microscope, floresencent microscope, and transmission electron microscope. The fluorescein isothiacyanate-dextran (FITC-Dextran) tracing technique was used to observe the inhibitory effect of the inclusion compound of curcumin on angiogenesis induced by colon carcinoma C26 cells in alginate beads.Results: The C-HP-?-CD and the mouse model of subcutaneous alginate-encapsulated colon carcinoma C26 cells were successfully created. Transmission electron microscope showed angiogenesis in alginate-encapsulated colon carcinoma C26 cells; claval endothelial cells were seen in some lumens. Optical microscope and the fluorescence microscope showed that the angiogenesis density of the positive group and curcumin groups were obviously lower than those of the negative control group; no obvious difference at vessel morphosis was found among the 3 alginate-encapsulated groups. FITC-Dextran tracing technology found that the fluorescent intensity of the negative control group was significantly higher than those of the positive control group and the curcumin group (P

Bovine leukemia virus(BLV)mainly causes bovine B-cell malignant lymphoma,which causes immunosuppression and leads animals susceptible to other diseases.BLV has been detected all over the world except in Western Europe.Antibodies to BLV have been detected in humans,and reverse transcribed DNA of BLV were detected in breast tissue sections and lung tumors,which in-dicate that BLV is a potential zoonotic disease.At present,there are no effective treatment methods or commercial vaccines for BLV.An in-depth understanding of the development of BLV detection technology will help researchers choose suitable detection methods,avoid invalid detection results,and meet the requirements of disease prevention and control.In view of this,this paper reviews 19 detection technologies of BLV including cytology detection,nucleic acid detection,and serology de-tection.The advantages,disadvantages,and scope of application were also analyzed,which provide theoretical guidance for the prevention and control of bovine leukemia.

Chicken coccidia specialize in growing and reproducing within the epithelial cells of the chicken intestinal mucosa,posing a threat to the health of the chicken flock.However,traditional chemical drugs are difficult to pass through the cell barrier and can easily lead to the emergence of drug-resistant strains and residual veterinary drugs,posing a huge challenge to the prevention and control of coccidiosis.Therefore,it is necessary to develop new strategies against coccidiosis.Nano-drugs have the advantages of good biocompatibility,easy modification,and high efficiency,impro-ving the penetration of cell membranes while reducing drug toxicity and side effects.They are ex-pected to be applied in the prevention and treatment of coccidiosis.This article introduces various nanomedicines for treating chicken coccidiosis,summarizes their advantages compared to tradition-al formulations,focuses on the application of nanomedicines in the treatment of chicken coccidio-sis,elucidates the existing problems of nanomedicine treatment for chicken coccidiosis,and looks forward to the challenges and future development directions in this field,in order to provide impor-tant references for the development of nanomedicines for treating chicken coccidiosis.