BACKGROUND:Bone marrow mesenchymal stem cel s and al ogenic bones are commonly used as seed cel s and scaffolds, respectively, for constructing tissue-engineered cartilage through in vitro co-culture. The loose body of the knee joint can survive in the articular cavity for a long time, and maintain certain characteristics of cartilage tissues. Therefore, the articular cavity can provide a good environment for the growth and development of chondrocytes. OBJECTIVE:To investigate the effect of bone marrow mesenchymal stem cel s co-cultured with al ogenic bone in the articular cavity. METHODS:Bone marrow mesenchymal stem cel s were isolated from five newborn New Zealand white rabbits. One adult New Zealand rabbit was enrol ed to prepare al ogenic bone for co-culture with bone marrow mesenchymal stem cel s. Afterwards, bone marrow mesenchymal stem cel s and al ogenic bone composites were cultured in the articular cavity (intracavitary culture group) or in vitro as in vitro culture group, respectively;the normal cartilage tissues grew in the articular cavity as control group. Cel s were observed by hematoxylin-eosin staining and type II col agen immunohistochemistry staining at 4, 8 and12 weeks of culture. RESULTS AND CONCLUSION:At 12 weeks culture, hematoxylin-eosin staining showed:in the control group, chondrocytes arranged tightly and directional y with red stained cytoplasm and cartilage matrix as wel as blue nuclei;in the intracavitary culture group, the scaffold was mostly absorbed and chondrocytes grew into the scaffold in a certain direction with smaller shape, while cytoplasm and cartilage ma trix were red stained, blue nuclei appeared;in the in vitro culture group, abundant chondrocytes proliferated in a disordered arrangement. Immunohistochemistry staining showed:the absorbance ( A) values in the intracavitary culture group showed a continuous increase, but no obvious change was in the other two groups. Moreover, at 4, 8 and 12 weeks of culture, A values in the control group and intracavitary culture group were significantly higher than that in the in vitro culture group (P<0.05);at 4 and 8 weeks, A value in the intracavitary culture group was significantly lower than that in the control group (P<0.05), but at 12 weeks, there was no significant difference in A value between the two groups (P>0.05). These results suggest that the tissue-engineered cartilage can be constructed by bone marrow mesenchymal stem cells co-cultured with allogenic bone under in vitro and in vivo environment, especially in the articular cavity.

Objective To investigate the method of screening metra scar flaw of late trimester of pregnancy with B ultrasound and predict the risk of scar uterine rupture. Methods The depth and scar of lower uterine segment were observed continuously by B ultrasound between 33～41 weeks of gravida having the history of cesarean section. Results Lower uterine segment has all been formed after 33 weeks of 396 gravida. Lower uterine segment flaw was found in 69 cases by B ultrasound, nine cases were diagnosed to be aura uterine rupture which were confirmed in operation. Conclusion With B ultrasound we can observe the development of lower uterine segment during pregnancy so as to predict aura uterine rupture.

Objective To evaluate the sensitivity of three different clinical consciousness scales to patients with disorders of consciousness.Methods A total of 84 patients in vegetative state (VS) were selected and scored by the consciousness scales including Glasgow Coma Scale (GCS),Coma Recovery Scale-Revised (CRS-R) and Chinese Vegetative State Scale (CVSS) respectively.The patients were followed every 2 weeks for 12 weeks or every 4 weeks for 12-24 weeks.The assessment finished in 6 months or till the time when patients emerged from minimally conscious state ( MCS.The diagnostic sensitivity and the sensitivities of each sub-scale in CRS-R were evaluated when the patients with MCS emerged from VS.The diagnostic consistency with other scales was observed and analyzed by Spearman correlation as well.The Spearman -relate was used to analyze the correlations among the total scores of three scales in the patients on admission (36 patients with acute and 48 patients with chronic phase).Results Of 84 VS patients,there were 47 emerged to MCS.CRS-R was the most sensitive in the diagnosis of MCS (P ＜ 0.05 ) among all scales.The visual subscale is the most sensitive among all the CRS-R subscales ( P ＜0.01 ).Among the 84 patients,the remaining 37 were diagnosed as VS by all three scales.The total scores correlation analysis showed that the score of CRS-R significantly correlate with that of GCS in acute and chronic phase ( P ＜0.01 ) and significantly correlate with that of CVSS only in the acute stage ( P ＜ 0.05 ).Conclusions It is indicated that CRS-R,especially the visual subscale of CRS-R is the more sensitive tool for detecting.MCS during recovery process in the disorders of consciousness.As to the patients with deep unconsciousness and with no significant recovery,the three instruments are in good agreement to determine the degree of disorders of consciousness.It is suggested that CRS-R can be widely used to assess the level of consciousness in various stages.

We aimed to examine the effect of pioglitazone on transdifferentiation of preosteoblasts from rat bone marrow mesenchymal stem cells (BMSCs) into adipocytes and investigate its effect on bone metabolism. BMSCs were harvested from the femurs and tibias of a rat, then separated, purified, proliferated for 3 generations and differentiated into preosteoblasts for 5 days and 14 days respectively in the presence of osteogenic medium. Thereafter, the preosteoblasts were cultured for 21 days in the presence of adipogenic medium with and without pioglitazone (1 μg/mL). Partially-differentiated osteoblasts were identified by mineralized nodules with Alizarin red S staining. Transdifferentiated adipocytes were identified by Oil Red O staining. Reverse transcription PCR (RT-PCR) was performed to assay the expression levels of osteogenic markers Runx2 and ALP, and an adipogenic marker PPARγ. Those cells cultured for 5 days did not show mineralized nodules as detected by staining of Alizarin red S, while those cultured for 14 days showed dispersed mineralized centers in the form of brown spots, although without obvious red mineralized nodules. After adipogenic transdifferentiation for 21 days, adipose-drops were found in cells of 5CG and 5EG earlier than those of 14CG and 14EG, and the former showed much more adipocytes separately as detected by Oil Red O staining. Whatever the time was 5 days or 14 days of BMSCs osteogenic differentiation, the cells cultured with pioglitazone showed much more adipocytes than those without pioglitazone. Our experiment showed that the less time it took for BMSCs osteogenic differentiation, a stronger ability remained for BMSCs to transdifferentiate into adipocytes. The mRNA expression levels of Runx2 and ALP were decreased by 1.79 and 1.90 times respectively in 5EG (P< 0.05) as compared with 5CG, and that of PPARγ was increased by 1.31 times in 5EG (P<0.05) as compared with 5CG. The mRNA expression levels of Runx2 and ALP were decreased by 1.45 and 1.54 times respectively in 14EG (P<0.05) as compared with 14CG, and that of PPARγ was increased by 1.39 times in 14EG (P<0.05) as compared with 14CG. It was concluded that pioglitazone stimulated the transdifferentiation of BMSCs into adipocytes. These observations provided a potential mechanism of imbalance in thiazolidinedione induced bone metabolism.

Objective To study a novel feature extraction method of Chinese materia medica (CMM) fingerprint. Methods On the basis of the radar graphical presentation theory of multivariate, the radar map was used to figure the non-map parameters of the CMM fingerprint, then to extract the map features and to propose the feature fusion. Results Better performance was achieved when using this method to test data. Conclusion This shows that the feature extraction based on radar chart presentation can mine the valuable features that facilitate the identification of Chinese medicine.

Objective To investigate treatment effect of the type of foot drop in patients with lumbar disc herniation.Methods Clinical data of 34 patients with foot drop in lumbar disc herniation who were treated in Spinal Surgery Department of Shunyi District Hospital of Beijing from January 2006 to December 2013 were retrospective analyzed,all patients were treated by operation.After 24 months of follow-up,the effect was observed,the prognosis and related factors were analyzed.Results Two years after operation,the recovery of the patients was excellent in 4 cases and good in 19 cases,common and poor in 11 cases,and the excellent and good rate was up to 67.6%(23/34).Analysis results showed that after 3 to 24 months after surgery,the anterior tibial muscle strength grades(grade 0-5) corresponding score were 28 points preoperatively,59 points at 3 months after surgery,69 points at 6 months after surgery,92 points 1 year after surgery,reached 95 points 2 years after surgery.Muscle strength score of patients with an increasing trend,especially after 3 months to 12 months after the operation.Conclusion The drop occurred should be operated as soon as possible,recommendations for open surgery,complete decompression,avoid injury.Three months to 1 years after operation is the key period of recovery.

Objective To study the variation of serum Hcy and UA of Parkinson's disease.Methods 46 pa-tients with Parkinson's disease in our hospital from April 2015 to March 2016 were selected as observation group,and 57 patients with other disease at the same time were selected as control group.The contents of serum Hcy and UA of the two groups were analyzed and compared.Results The contents of serum Hcy and UA of the observation group were (17.82 ±3.12)μmol/L,(321.42 ±79.42)μmol/L.Those of the control group were (10.12 ±3.05)μmol/L, (420.15 ±70.18)μmol/L.The content of serum Hcy of observation group was higher than the control group,the con-tent of serum UA of the observation group was lower than the control group(t =12.608,P <0.01).Hcy content of middle and late patients[(18.86 ±3.12)μmol/L]was higher than early patients[(16.16 ±3.04)μmol/L](t =2.378,P <0.05),serum Hcy level was positively correlated with UPDRS(r =0.462,P =0.037),the differences were significant.Conclusion Changes of serum Hcy and UA levels were related to the occurrence of Parkinson's disease.

Objective To investigate the effect of metformin on proliferation and differentiation of bone marrow mesenchymal stem cells (MSCs) cultured in adipogenie medium. Methods Bone marrow MSCs were isolated from SD rats and cultured in adipogenic medi- um with or without 100?mol/L metformin. MTT test was performed to evaluate the proliferation of cells, while oil red O staining and real time RT - PCR were performed to evaluate the differentiation of cells. Results The number of bone marrow MSCs in two groups both increased over time, and metformin induced more cells. However, the cells affected by metformin showed smaller and less cytoplasmic lipid droplets compared with the cells in control group. Moreover, real time RT - PCR has shown the inhibitory effect of metformin on adipocyte differentiation, with significantly decreased mRNA levels for adipogenic markers. Conclusion Metformin may promote proliferation, but inhibit differentiation of bone marrow MSCs cultured in adipogenic medium.

Objective To determine MIF expression in normal colorectal mucosa,colorectal adenoma and colorectal carcinoma, and the relationship between MIF and the clinical pathologic features of colorectal carcinoma.MethodsMIF expression was determined by immunohistochemical staining ELISA was used to detect the level of MIF in serum samples. Results The percentage of MIF expressing epithelial cells and MIF positive expression intensity were progressively increased in normal colorectal mucosa, colorectal adenoma and colorectal carcinoma. There was significant difference among three groups (P

Objective In this study,we elucidated the role of high mobility group box chromosomal protein 1 (HMGB1) in collagen-induced arthritis (CIA) rat and the antagonist role of ethyl pyruvate by using a rat model of CIA as the research object by comparing the expression of HMGB1 in normal control group,CIA model group and ethyl pyruvate group.Methods Thirty-six rats were randomly divided into 3 groups (n=12):normal control group,CIA group and ethyl pyruvate group.Then the 6 rats were dissected at the 6th,9th week respectively.Thc expression of HMGB1 was analyzed by immunohistochemistry and Pathology-image analysis software in the cytoplasma.The expression of HMGB1 mRNA with real time-polymerse chain reaction (PCR) was evaluate,and the HMGB1 expression of each group were compared with t-test.Results The immunohistochemical results of HMGB1 showed that the expression intensity in the normal control group,CIA model group and ethyl pyruvate group was 2.1±0.6,7.3±1.2,6.0±1.2 respectively at the 6th week; and 2.2±0.7,12.4±4.5,5.5±1.0 at the 9th week respectively.The HMGB1 mRNA real time-PCR results had shown that the relative quantification of the normal control group and CIA model group were 1,2.865,2.602respectively at the 6th week and 1.005,4.694,1.729 at the 9th week.At those two points, the HMGB1 expressions of HMGB1 antagonist group were significantly higher than those of the normal controls (P＜0.05).In addition,there was statistical significant difference(P＜0.05) in the HMGB1 expression when compared with the placebo group.Furthermore, when the degree of HMGB1 expression among the three groups was compared,the HMGB1 antagonist group was decreased significantly (P＜0.05).Conclusion The results has demonstrated that HMGB1 could induce inflammation in the synovial tissue of CIA rats,and has provided the rationale that HMBG 1 could be the target of treating rheumatoid arthritis (RA).The results of this study have shown that ethyl pyruvate could antagonize the effect of HMGB1.This finding may provide a new therapeutic target for the treatment of RA.