BACKGROUND: There are different methods to isolate and culture human nucleus pulposus cells, and the differences in digestive enzymes components and digestion time quite are significant. So how to rapidly and efficiently harvest human nucleus pulposus cells has become a research hotspot. OBJECTIVE: To optimize the digestive enzymes components and digestion methods for the preparation of human nucleus pulposus cells. METHODS: Nucleus pulposus tissue specimens were selected from three adult discs in the Department of Orthopedics, China-Japan Union Hospital of Jilin University. The acute traumatic disc tissues that outstanding to the spinal canal were taken under aseptic conditions, and then the peripheral white annulus and jel y-like nucleus pulposus in the center could be seen. According to different mixed enzyme concentration ratio, the samples were divided into two groups. The enzyme Ⅰ group was treated with 0.2% Ⅱ col agenase; and the mixed enzymeⅡ group was digested with 0.25% trypsin for 30 minutes, and then treated with 0.2% Ⅱ col agenase. According to digestion time, each group was divided into three subgroups: 2 hours group, 4 hours group, and overnight group. Final y, suspended cel volume was decided as 2 mL to count cells. Dulbecco’s modified Eagle’s medium containing fetal bovine serum was used for cel culture in vitro. Trypan blue staining was performed to count total cel number and ratio of living cells. Methylthiazolyldiphenyl-tetrazolium bromide assay was used to detect the growth curve of nucleus pulposus cells. RESULTS AND CONCLUSION: Based on the two digestion enzyme concentration, the number of digested cells in the enzyme Ⅰ group was larger than that in the enzyme Ⅱ group after digested for 2 and 4 hours, but the difference was not significant (P > 0.05). Overnight, cellsurvival rate was decreased in the enzyme Ⅰ group after digested for 2 and 4 hours when compared with the enzyme Ⅱ group, and the difference was significant (P < 0.05). After digested for 4 hours, tissue blocks disappeared, and the number of cells reached maximum. The results indicate that enzyme Ⅰgroup composite with Ⅱ col agenase is benefit for the separation of nucleus pulposus cells, and the digestion time is appropriate to 4 hours. This condition has the advantages of simple operation, high efficiency and low cost, and it considered that digestion of nucleus pulposus tissues with 0.2% Ⅱ col agenase for 4 hours is the best condition to obtain nucleus pulposus cells.

BACKGROUND:Advancement in bioengineering based upon tissue engineering techniques may offer the possibility of repairing degenerative intervertebral disc.
OBJECTIVE:To summarize the research progress in the scaffolds of tissue engineered intervertebral disc.
METHODS:A computer-based retrieval was performed to search manuscripts describing tissue engineered intervertebral disc scaffolds published between January 1st, 1900 and December 31st, 2012 in PubMed database with the key words of“tissue engineering, intervertebral disc, scaffold”in English.
RESULTS AND CONCLUSION:Scaffold is an important part of tissue-engineered research. There are three kinds of materials for intervertebral disc scaffolds:natural biomaterials, synthetic materials, and composite materials. A variety of scaffold materials have their own advantages and disadvantages. Up to now, none of these scaffold materials is accepted as the most suitable one. The selection of scaffold materials is stil to be further studied. The study and development of nanoscale biomaterials is an inevitable trend. Otherwise, with the help of bionics, improving scaffolds is also an inexorable trend in progress of simulating human intervertebral disc. Furthermore, injectable scaffold is also an research hot spot, and the selection range of injectable scaffold materials mainly focuses on chitosan, typeⅡcolagen,hyaluronic acid,fibrin,elastin,and alginate.C urrently, studies on chitosan as a scaffold material are relatively more.

Objective To investigate prevalence of nutritional status of children and adolescents of school age in the poor region with many ethnic groups of Western regions of China. Methods In September 2006, we performed a health check?up for 2 475 students of a primary school and a middle school from many ethnic groups in Rongshui, Guangxi; 1 163 boys (46.99%) and 1 362 girls (53.01%) participated in this study, including 4 main ethnic groups: Han (848 participants, 34.26%), Miao (736 participants, 29.74%), Zhuang (415 participants, 16.77%) and Dong (341 participants, 13.78%), and prevalence of obesity and malnutrition for the population were surveyed. Results The prevalence rates of overweight among male and female children/adolescents of school age were 3.01%and 3.66%, respectively;and the prevalence rates of obesity were 1.55%and 0.76%, respectively. In 4 main ethnic groups of Han, Miao, Zhuang and Dong, the prevalence rates of overweight were 2.59%, 4.76%, 2.89% and 2.93%, respectively; while the prevalence rates of obesity were 1.89%, 0.27%, 0.96%and 1.17%, respectively. The prevalence rates of malnutrition among male and female children/adolescents of school age were 5.59%and 3.35%, respectively;while the prevalence rates of malnutrition in 4 main ethnic groups of Han, Miao, Zhuang and Dong were 6.96%, 2.17%, 5.54% and 2.64%, respectively. Conclusion The prevalence rates of overweight and obesity of children and adolescents of school age in Rongshui were relatively low, while the prevalence rates of malnutrition is similar to the average level of China.

Objective To identify the applicability of Traditional nutritional evaluation method,short-form mini-nutritional assessment (MNA-SF) and nutrition risk screening 2002 (NRS 2002) on screening malnutrition in elderly in-patients.Methods 100 elderly in-patients with malnutrition were screened by the above screening instruments.The evaluation effects of the 3 methods were compared by sensitivity,specificity and ROC curve.Results The detection rates of elderly in-patients with malnutrition by BMI,MNA-SF and NRS 2002 was 7％,58％ and 39％ respectively.Taking the human body measurement instrument as standard,the sensitivities of BMI,MNA-SF and NRS 2002 was 0.475,0.918 and 0.410respectively,the specificities was 0.615,0.103 and 0.641 respectively.3 tools series test sensitivity and specificities was 0.557 and 0.913 respectively,parallel test sensitivity and specificities was 0.929 and 0.435 respectively.Conclusions There is significant difference among the sensitivities of the 3 methods,NRS 2002 has the highest accuracy and is applicable to the screening of nutrition risks in elderly patients.Combined 3 kinds of tool can better improve the effect.

Objective Detecting antibodies against RBCs,platelets,lymphocytes,and neutrophils by a solid-phase antibodies screening system.Methods mono-layer blood cell immobilized in the bottom of U-microplate respectively to prepare the solid-phase antibody screening system.Then detecting the antibodies exist or not in 2 150 random blood donor and 440transfusion patients' samples.Analyze the influence of antibodies against blood cells to the transfusion effect.Results 53RBC antibodies,22 platelet antibodies,13 lymphocyte antibodies,55 neutrophil antibodies were detected in random blood donor samples;31 RBC antibodies,38 platelet antibodies,24 lymphocyte antibodies,43 neutrophil antibodies and 115 mixed (two or more antibodies against RBC,platelets,lymphocytes or neutrophils exist at the same time) were detected in transfusion patients' samples after detection.233 suspected adverse reaction happened after transfusion,which antibodies were detected in 133 samples.Conclusion The antibodies against blood cells solid-phase screening system can applied in pre-and post-transfusion detection.The solid-phase screening method is more sensitivity than serologic tube test and flow cytometry.Existing of antibodies influence the transfusion effect.

OBJECTIVE: To establish a supplementary diagnostic indicator (infrared radiant intensity) in tongue diagnosis of traditional Chinese medicine (TCM) in patients with hyperplasia of mammary glands through correlation analysis of infrared radiant intensity between hyperplastic breast tissue and tongue surface. METHODS: Infrared radiant intensity of the hyperplastic breast tissue and different points on tongue surface in 20 cases of hyperplasia of mammary glands with liver-energy stagnation and phlegm retention syndrome and 16 cases of hyperplasia of mammary glands with irregular thoroughfare and conception vessels syndrome were measured with external infrared spectrometer PHE-201 made by Shanghai Institute of Technical Physics. Correlation of infrared radiant intensity between the hyperplastic breast tissue and the different points on tongue surface was assessed by using bivariate correlation analysis. RESULTS: The results showed that the numbers of positive correlated wave bands of infrared radiant intensity between the hyperplastic breast tissue and different detected points on tongue surface in the patients with liver-energy stagnation and phlegm retention syndrome and irregular thoroughfare and conception vessels syndrome were 127 (83.55%) and 71 (46.71%), respectively. Infrared radiant intensity between the hyperplastic breast tissue and the tongue surface had a positive correlation. CONCLUSION: Infrared radiant intensity can be used as one of supplementary diagnostic indicators in TCM tongue diagnosis of hyperplasia of mammary glands.

To study the complete genomic sequence, genomic characteristics, and genetic variation of the bovine papillomavirus 2 genotype (BPV-2) Aks-01 strain at the molecular level, genotyping of this strain from the skin samples of cows in southern Xinjiang (China) was first detected by the polymerase chain reaction with FAP59/FAP64 primers. Based on the complete genome of the BPV-2 reference strain, specific primers and sequencing primers were designed, and the complete genome of the Aks-01 strain amplified and sequenced. Sequence analyses showed that genotyping of the Aks-01 strain belonged to BPV-2. The Aks-01 strain had the structural characteristics of BPV-2. The 7944-bp full-length genomic sequence of the Aks-01 strain was compiled using DNAStar™. The sequence of the Aks-01 strain had 98% similarity to the reference strain from GenBank. The Aks-01 strain was most closely related to BPV-1 and BPV-13. BPV-2, BPV-1 and BPV-13 were grouped within the genus Deltapapillomavirus. The Aks-01 strain is the first BPV-2 strain reported in southern Xinjiang.
Amino Acid Sequence ; Animals ; Base Sequence ; Bovine papillomavirus 1 ; genetics ; Cattle ; China ; Evolution, Molecular ; Female ; Genome, Viral ; genetics ; Genomics ; Genotype ; Molecular Sequence Data ; Oncogene Proteins, Viral ; chemistry ; genetics ; metabolism ; Phylogeny ; Sequence Analysis, DNA ; Skin ; virology

Objective To investigate the clinical characteristics and the experience of multi-disciplinary team (MDT) on recurrence of primary hyperoxaluria (PH) type I after renal transplantation. Methods One case presenting with unexplained rapid decline of renal allograft function after allogeneic renal transplantation was discussed by MDT. The role of MDT in diagnosing rare hereditary diseases and improving the long-term survival of renal transplant recipients was summarized. Results After MDT consultation, the patient was diagnosed with recurrence of PH type I. Routine immunosuppressive regimen was initiated after the exclusion of rejection. The patient was instructed to drink a large quantity of water, and given with high-quality protein and low-phosphorus diet, vitamin B6, calcium and other conservative therapies to actively prevent and treat postoperative complications. The deterioration of renal graft function was delayed. Nevertheless, regular hemodialysis was resumed at 5 months after renal transplantation until the submission date of this manuscript. Conclusions Recurrence of PH type I after renal transplantation is relatively rare. The main clinical manifestations are recurrent kidney stones and decreased renal function with multiple complications and poor prognosis. The condition of the patient is consulted by MDT for confirming the diagnosis, determining the optimal treatment scheme, delaying the progression and improving the clinical prognosis.

Objective:To explore the early response of hepatobiliary biochemical indexes after short-term ursodeoxycholic acid treatment in patients with primary biliary cholangitis (PBC).Methods:According to the Child-Pugh (C-P) score, the patients with newly diagnosed PBC were divided into groups A, B and C. The early biochemical response was defined as the improvement of hepatobiliary biochemical indexes while daily dose of 13-15 mg/kg ursodeoxycholic acid(UDCA) was administrated for 3-4 weeks. Rank sum test was used to compare the alkaline phosphatase (ALP), γ-glutamyl transpeptidase (GGT), aspartate aminotransferase (AST), total bile acid (TBA) and total bilirubin (TBil) before and after treatment between groups. Chi-square test was used to analyze the differences in numerical data between groups. ROC curve was also used for data an-alysis.Results:The levels of AST, ALP, GGT and TBil of groups A and B after treatment were lower than those of before treatment ( P<0.05). And there was no significant difference in group C ( P>0.05). On the contrast, TBA levels after treatment in group A, B and C were significantly higher than those of before ( P<0.05). ROC curve analysis of the baseline ALP, GGT and AST indexes that declineed less than 20% showed that their cut-off value of indexes were 211.00, 285.85 and 86.68 U/L respectively in group A. And then the cut-off value of above parameters in group B was 505.00, 353.10 and 179.15 U/L respectively. But no statistical significant difference was found in ROC curve analysis of above indexes of group C. The baseline TBil level declined synchronously with the decline of ALP, AST and GGT less than 20% were analyzed by ROC curve, but no statistical significant difference was found in group A, B and C. ROC curve analysis of the treatment showed that increased of TBA level was synchronously to the decline of ALP, AST and GGT less than 20% showed that cut-off value were 38.75 and 35.95 μmol/L respectively in group A and B. There was no statistically significant difference in ROC curve analysis of TBA in group C. As for baseline ALP, GGT and AST their level were decreased with treatment, and less than 40%, ROC curve analysis did not find statistical significant difference in above indexes. Conclusion:After 3-4 weeks of UDCA treatment, the cut off values of ALP, GGT and AST in Grade A or B of C-P are met, and the biochemical response could be reduced by 20% or more, among which the accuracy of ALP is higher. The TBA level of C-P in grade A or B PBC patients after UDCA treatment increases, and when the Cut-off value is met, it suggests that ALP, GGT and AST indexes may decrease by 20% or more. Total bilirubin does not show response to treatment.

Objective:To summarize the clinical characteristics of central nerve system (CNS) infection and grasp the necessity and possibility of early diagnosis and precise intervention of CNS infection after renal transplantation.Methods:This retrospective study enrolled consecutive recipients of renal transplantation with CNS infection after transplant between January 2000 and December 2020. Correlative factors for CNS infection after renal transplant were determined by comparing the clinical data between recipients with and without CNS infection. After screening 3, 199 consecutive renal transplant recipients, 12 patients with CNS infection post-transplant were identified and recruited. The median age-of-onset was 48.5 (23-65) years. And the median time to disease onset after transplant was 50.5(1-204) months. The most common symptoms of CNS infection after renal transplant included fever (75.00%), consciousness disorder (58.33%), headache (58.33%) and neck rigidity (41.67%).Results:Hepatitis B virus carrier and pulmonary infection were correlated with CNS infection after transplantation ( P<0.05). Nine patients failed to identify the pathogen and only received empirical anti-infective regimen. The outcomes were curing ( n=3) and death ( n=6). Metagenomic sequencing was performed for identifying the pathogen in three recipients and actively adjusting the anti-infective regimen. As a result, 2 were cured and 1 died. The overall mortality was 58.33%. The median time to death or curing from disease onset were 20(2-19) and 25(16-35) days respectively in surviving and non-surviving recipients. Conclusions:The progress of CNS infection after transplantation is rapid with a high mortality. HBV carrier and pulmonary infection are possible risk factors of CNS infection after renal transplantation. Early pathogenic identification and precise etiological intervention are vital for better clinical outcomes.