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.

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 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

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 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.

OBJECTIVETo investigate the clinicopathologic characteristics, prognosis and histologic origin of the mucinous tumor of the peritoneum.

METHODSAccording to 2010 WHO classification of tumours of the digestive system, 34 cases diagnosed as "pseudomyxoma peritonei (PMP) " were reevaluated and divided into low grade and high grade. Immunohistochemistry was applied to investigate the expression of SATB2 and the histologic origin of the mucinous tumor of the peritoneum, using antibodies against SATB2, CK7, CK20 and CDX-2. The relationship between clinicopathologic characteristics and prognosis of the low grade and high grade tumors were analyzed.

RESULTSTwenty five patients had low grade mucinous tumors (two of them were no cell type), nine patients had high grade mucinous tumors. There was no significant difference between low grade and high grade mucinous tumors in age, sex, recurrence and organs involvement (P>0.05). Thirty patients were followed up, the overall survival rates of patients with low grade and high grade mucinous tumors were 13/21 (61.9%) and 3/9, respectively. The median survival time was 74 and 24 months in low and high grade patients, and the difference was statistically significant (P=0.002).Immunohistochemistry showed the expression rates of CDX-2, CK20, and CK7 in totally 32 cases (excluding 2 cases of no cell type) were 30/32(93.8%), 31/32 (96.9%), and 3/16, respectively; the expression rates of CDX-2, CK20, and CK7 in 16 cases with distinct primary site were 15, 16, and 1, respectively; fifteen of 16 cases of tumors of unknown primary site were positive for CDX-2 and CK20, two of the them were positive for CK7. There was no difference in the expression of CDX-2, CK20 and CK7 between tumors with distinct primary site and tumors with unknown primary site (P>0.05). The expression rate of SATB2 in the cases was 56.3% (18/32), excluding 2 cases of no cell type. There was no significant difference between low grade and high grade tumors in the expression of SATB2 [15/23(65.2%) vs 3/9, P=0.102], also SATB2 was not related to the prognosis of the tumor (P=0.786).

CONCLUSIONThe prognosis of the mucinous tumor of the peritoneum was significantly different between low grade and high grade according to WHO 2010 classification, and most mucinous tumor of the peritoneum originated from the appendix.

Adenocarcinoma, Mucinous ; metabolism ; pathology ; secondary ; surgery ; Adult ; Aged ; Aged, 80 and over ; Appendiceal Neoplasms ; pathology ; surgery ; CDX2 Transcription Factor ; Female ; Follow-Up Studies ; Homeodomain Proteins ; metabolism ; Humans ; Keratin-20 ; metabolism ; Keratin-7 ; metabolism ; Lymphatic Metastasis ; Male ; Matrix Attachment Region Binding Proteins ; metabolism ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; Peritoneal Neoplasms ; metabolism ; pathology ; secondary ; surgery ; Pseudomyxoma Peritonei ; metabolism ; pathology ; surgery ; Survival Rate ; Transcription Factors ; metabolism

OBJECTIVE: To evaluate the prospective association between cumulative resting heart rate (cumRHR) and rapid renal function decline (RRFD) in a cohort of individuals aged 60 and older. METHODS: In the Tianjin Chronic Kidney Disease Cohort Study, the individuals who underwent three consecutive physical examinations between 2014 and 2017, with estimated glomerular filtration rate (eGFR) greater than 60 mL/min per 1.73 m² and aged 60 years or older were enrolled. A total of 27,564 patients were prospectively followed up from January 1, 2017 to December 31, 2020. The 3-year cumRHR was calculated. The primary outcome was RRFD, defined as an annualized decline in eGFR of 5 mL/min per 1.73 m² or greater. Logistic and restricted spline regression models and subgroup analysis were used to investigate the association of cumRHR with RRFD after adjusting for all confounders. RESULTS: During a median follow-up of 3.2 years, a total of 4,347 (15.77%) subjects developed RRFD. In fully-adjusted models, compared with the lowest quartile of cumRHR, the odds ratio (OR) for the highest was 1.44 (1.28-1.61), P < 0.001. Furthermore, each 1-standard deviation (27.97 beats/min per year) increment in cumRHR was associated with a 17% (P < 0.001) increased risk of RRFD, with a linear positive correlation (P for non-linear = 0.803). Participants with a 3-year cumRHR ≥ 207 (beats/min) * year (equivalent to ≥ 69 beats/min per year in 3 years) were found to be at a higher risk of RRFD. CONCLUSIONS The cumRHR is significantly associated with a higher risk of RRFD among older adults. These results might provide an effective goal for managing and delaying the decline of renal function in the older adults.