BACKGROUND: In the field of intervertebral disc tissue engineering, seed cells primarily consist of nucleus pulposus cells (NPCs) and mesenchymal stem cells (MSCs). NPCs have been known to have several shortcomings: limited source, inconvenient collection, poor proliferative capacity, and difficult in vitro culture.OBJECTIVE: To investigate the proliferation of MSCs after co-culture with NPCs in alginate beads-simulated three-dimensional environment.DESIGN, TIME AND SETTING: A cell observation was performed at the Laboratory of Orthopedics, Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology between December 2007 and October 2008.MATERIALS: Six healthy New Zealand rabbits of 4 months old were provided by the Laboratory Animal Center, Tongji Medical College of Huazhong University of Science and Technology and were used for the present study.METHODS: Rabbit MSCs were isolated and purified using density gradient centrifugation and adherence methods. Rabbit NPCs were isolated and purified using collagenase Ⅱ digestion and adherence methods. Following liposome-mediated green fluorescent protein tranfection and G418 screening, MSCs of passage 3 were cultured either alone (control group) or with NPCs at a ratio of 1:1 (co-culture group) in alginate beads. After 14 days of culture, alginate beads were dissolved and MSCs were collected by flow cytometry sorting.MAIN OUTCOME MEASURES: The expression of collagen Ⅱ and aggrecan in MSCs was detected by RT-PCR and Western blot techniques.RESULTS: mRNA and protein expression of collagen Ⅱ and aggrecan was observed in the co-culture group, but not in the control group, after 14 days of culture.CONCLUSION: MSC differentiation towards nucleus pulposus-like cells can be induced by co-culture with NPCs in a three-dimensional environment.

To study xanthones from the barks of Garcinia xanthochymus, the constituents were isolated by normal-phase and reverse-phase silica gel column chromatography from the EtOAc extract. Their structures were elucidated by spectral analysis. Three new xanthones were purified and identified as 1,2,5-trihydroxy-6-methoxyxanthone (1), 1,4,6-trihydroxy-5-methoxyxanthone (2), 1,2,7-trihydroxy-4-( 1,1-dimethylallyl) xanthone (3).

Targeted therapeutic strategy for cancer stem cells (CSCs) is the key to prevent tumor relapse and metastasis. The im-portant roles of epigenetic regulation on the development of stem cells and gene reprogram of somatic cells suggest that this process may remarkably affect the occurrence and development of CSCs. The epigenome, which comprises DNA methylation, histone modifica-tions, chromatin structures, and non-coding RNAs, controls gene expression patterns. In renal cell carcinoma (RCC), aberrant changes occur in the epigenome. To date, cells with CSC properties from RCC have been successfully isolated using different methods, such as sorting using the Hoechst 33342 side population, forming tumor spheroid, and sorting CD105 cell surface biomarker. According to the progress in genetic studies on RCC, in addition to DNA sequence, the abnormality in the regulatory mechanism has considerable func-tions in tumor progression. Epigenetic changes may be integral to the behavior of cancer progenitor cells and their progeny. Knowledge on epigenetics in renal tumorigenesis process is beneficial in the development of new therapeutic modalities and may deliver new prog-nostic and early diagnostic markers. This paper reviews the latest development in the study of RCC stem cells and the underlying mech-anisms of epigenetic regulation on the development of CSCs in RCC.

Objective To assess the effects of eszopiclone (ESZ) on the pentobarbital sodium-induced sleeping time and spontaneous activity in mice exposed to acute hypobaric hypoxia.Methods 120 mice were randomly divided into 6 groups by using two factors 2×3 levels factorial design,in which two factors were interventions (ESZ and 0.9％ sodium chloride,2 levels) and altitudes (800 m,3500 m and 6000 m,3 levels).The pentobarbital sodium-induced sleeping test and the open field test were engaged to assess the effects of ESZ on sleeping time and spontaneous activity.Results (1) The drug and altitude had no interaction in the results of both the pentobarbital sodium-induced sleeping test and the open field test(P＞0.05).(2)The time of pentobarbital sodium-induced sleeping of mice in the groups of ESZ at each altitudes were (37.77± 18.22) min,(37.02± 13.67) min,(95.67±47.68)min and in the groups of NS were(17.78± 14.10) min,(15.09± 12.46) min,(39.54±28.24) min respectively,and the sleep time in ESZ groups were significantly longer than those in the groups of NS (P＜0.05).The time of pentobarbital sodium-induced sleeping were longer in group of 6000 m than those in the other two groups,both the ESZ and NS groups (P＜0.05).(3)No significant difference was found in the open field test between the ESZ and NS groups in the same altitude(P＞0.05) ; while the mice at the altitude of 6000 m in groups of ESZ and NS decreased compared with the groups at the altitude of 800 m after the relevant drugs intra-perineally for 6 h (P＜0.05).Conclusion ESZ may prolong pentobarbital sodium-induced sleeping time especially at the altitude of 6000 m and with no influence on the spontaneous activity in mice exposed to acute hypobaric hypoxia.High altitude at 6000 m may prolong the sleep time induced by pentobarbital sodium and reduce the spontaneous activities.

Objective To evaluate the mid-term results of arthroscopic anterior cruciate ligament (ACL) reconstruction using Transfix technique.Methods From August 2002 to December 2003,38 patients (24 left knees and 14 right knees) with ACL ruptures underwent arthroscopic ACL reconstruction with hamstring tendon using Transfix femoral fixation and interference screws tibial fixation.There were 21 males and 17 females,aged from 19 to 48 years (average,28.4 years).The causes of injury included sports injury in 27 cases,traffic injury in 2 cases and fall injury in 2 cases,and 7 patients had no significant injury.There were 6 cases of acute injury and 32 cases of old injury.Anterior drawer test was positive in 35 cases,weak positive in 1 case and negative in 2 cases.Lachman test was positive in 37 cases,and weak positive in 1 case.The Lysholm score was used to evaluate the results.Moreover,X-rays and MRIs were used to observe the conditions of graft and bone tunnel.Results Thirty-six patients were followed up for 6.3 to 7.6 years (average,6.8 years).All patients had normal joint range of motion.The Lysholm score improved from preoperative 64.4±4.52 to postoperative 85.6±4.60,and the difference was statistically significant (P ＜ 0.05).The X-rays and MRIs showed femoral and tibial tunnel enlargement in 3 cases,femoral tunnel enlargement in 5 cases and tibial tunnel enlargement in 3 cases.No joint space narrowing was found in all patients.One patient underwent arthroscopic revision ACL reconstruction with allograft tendon using bioabsorbable interference screws fixation on both tibial and femoral sides for re-rupture of ACL 4 years after operation due to trauma.Conclusion Arthroscopic ACL reconstruction with hamstring tendon using Transfix femoral fixation and interference screws tibial fixation is an effective method,which can provide satisfactory joint range of motion and stability.

Objective To prepare the chitosan nanoparticles loading 5-[125Ⅰ] Iodo-2'-deoxyuridine (125Ⅰ-UdR-CS-DLN) at 100-200 nm in diameter, and analyze the characteristic of drug sustained-releasing and tumor targeting. Methods Orthogonal experimental design and One-way analysis were applied to optimize the preparation of 125Ⅰ-UdR-CS-DLN using tripolyphosphate cross-linking. Dynamic dialysis was utilized to investigate the in vitro releasing characteristics of the nanoparticles. The tumor targeting effect of the nanoparticles was observed with laser confocal microscopy. Results The optimal conditions for preparing the nanoparticles at particle diameters (70. 39 ± 5.12 ) nm (PDI 0. 16 ± 0. 012 ) were 1 g/L of CS, 2 g/L of TPP, stirring rate 600 r/min, relative molecular mass of CS 3 × 103. The TEM results showed that the exterior of the nanoparticles was spheroid, with a uniform and fine dispersivity. The optimized condition with the initial 125Ⅰ-UdR concentration of 2. 96 MBq/ml at pH5 provided the highest loading capacity (1253. 55 MBq/g) and the highest entrapment rate (42. 35% ). The in vitro releasing curves of 125Ⅰ-UdR-CS-DLN followed Higuchi equation, shown a characteristic of long-acting preparation.Laser confocal microscopy observations approved that the tumor cells uptake of FITC-CS-nanoparticles were significantly more than that of normal cells. Conclusions Chitosan nanoparticles loading 125Ⅰ-UdR at diameters range 127. 81 ± 15. 25 nm (PDI 0. 240 ± 0. 035 ) were successfully prepared with the optimized conditions, and showed a characteristic of sustained-releasing and tumor targeting. The chitosan-based nanotechnology provided a new and efficient approach for the application of 125Ⅰ-UdR in intracellular radiotherapy for tumor.

Objective To evaluate the value of computed tomography(CT) and magnetic resonance imaging(MRI) in diagnosis of avascular necrosis of the femoral head(ANFH). Methods The literatures of ANFH diagnosed with CT and MR imaging published in the last ten years were collected by searching. Of that,21 literatures were correspond for the standards in this study and were select-ed. These literatures in diagnosing ANFH with CT and MR imaging were analysed with Meta-analysis by the sofeware of StataSE10.1. Results MRI was more effective than CT in diagnosing ANFH. There was significantly different in statistics between them [OR=0.13,95%CI(0.03～0.51)]. Conclusion In comparison with CT,MRI is the better method in diagnosing ANFH.

0.05).The differences of comprehensive scoring of QOL and scoring of factors 1,2,3 and 4(F_1,F_2,F_3 and F_4)in group A were significant as compared with those before treatment(P0.05),indicating that oral use of SRTT inhibited the aggravation of pulmonary function.The improvement of QOL in group A was superior to that in group B after treatment and the difference was significant(P

AIM:To explore the application effect of pranoprofen combined with tobramycin and dexamethasone ophthalmic suspension in cataract extraction after IOL implantation.METHODS:Retrospective analysis of the clinical data of cataract patients treated from May 2015 to June 2016 in our hospital.According to the treatment methods, patients were divided into tobramycin and dexamethasone ophthalmic suspension with pranoprofen group (combined treatment group) and pranoprofen treatment group.The difference of visual acuity, intraocular pressure, anterior chamber flash change, satisfaction rate and symptom score before and after treatment in two groups were observed.RESULTS:The vision of the two groups before treatment had no difference(P>0.05).At 1wk and 1mo after treatment, the visual acuity of the two groups all improved compared with before treatment, there was no significant difference between the two groups (P>0.05).Before treatment, the anterior chamber flare of the two groups had no difference(P>0.05).At 1wk and 1mo after treatment, the anterior chamber flare of combined treatment group was lower, the difference was statistically different (t=2.435, 1.864;P<0.05).The symptom score of the two groups before treatment had no difference(P>0.05).After treatment, the symptoms and signs of combined treatment group was significantly lower than that of pranoprofen group (t=2.586, 7.820;P<0.05).The satisfaction rate of the combined treatment group patients was 100%, significantly higher than pranoprofen group.CONCLUSION:Pranoprofen and tobradex in the treatment of cataract extraction and intraocular lens implantation has good application effect, can significantly improve the patient's symptoms and signs.

Objective　To approach the relationship betwe en glycosaminoglycan metabolism in cartilages and pathogenesis of KBD.Methods　Rhesus monkey was fed with grains and water from KBD endemic area for 18 months to produce the animal model with KBD . The glyc osaminoglycans in the monkey cartilage were extracted by the improved Dish method of Bitter. Purified glycosaminoglycans were digested with chondroitinaseABC, and the enzymatic digests were analyzed by HPLC. Results　Comparing with those of the control, the glycos aminoglycans in the head of femur, tibia plateau and costal cartilage from the Rhesus monkey fed with grains and water from KBD endemic area were undersulfated . Decreased unsaturated 4-sulphated disaccharide (△Di-4S) from the glycosa minoglycans in the head of femur and tibia plateau and decreased unsaturated 6- sulphated disaccharide (△Di-6S) from the glycosaminoglycans in the costal cart ilage were discovered.Conclusions　Detrimental factors in grains and water from KBD endemic area cause undersulfate of the cartilages glycosaminoglycans from Rhesus monkey. The glycosaminoglycans changes have a direct bearing on the patho logical alterations in morphology on the cartilages from the animal model with K BD.