BACKGROUND:Inducing factor and chondrogenic microenvironment is a primary factor, which influences chondrogenic differentiation and chondrogenesis of bone marrow-derived mesenchymal stem cells (MSCs). OBJECTIVE:To explore the feasibility of in vivo chondrogenesis by co-culture of bone marrow-derived MSCs and chondrocytes. DESIGN, TIME AND SETTING:A randomized controlled animal experiment was performed at Department of Pathology, Stomatological Hospital, Fourth Military Medical University of Chinese PLA between September 2004 and March 2005. MATERIALS:Fifteen New Zealand rabbits of clean grade were used for cell-scaffold construct transplantation. The rabbits were randomly divided into co-culture, chondrocyte, and bone marrow-derived MSC groups, with 5 rabbits in each group. Five neonatal New Zealand rabbits, aged 1-3 days, were used for isolation and culture of bone marrow-derived MSCs and chondrocytes. Polyglycolic acid (PGA) scaffold material (Shanghai Yikuo Company, China) has a fiber diameter of 15 μm, with an average interval of 150-200 μm, an interval porosity of 97% and 2-mm thickness. METHODS:In the co-culture group, bone marrow-derived MSCs and chondrocytes were mixed at a ratio of 3:1. The mixed cells were seeded onto a pre-wetted PGA scaffold (5 mm×5 mm )at the ultimate concentration of 6.0×1010 L-1. Dulbecco's modified Eagle's medium (DMEM) supplemented with fetal bovine serum was dropwise added to peripheral compound for 1 week of culture. In the chondrocyte, and bone marrow-derived MSC groups, chondrocytes and bone marrow-derived MSCs of the same ultimate concentration were seeded respectively onto the PGA scaffold. Then, the cell-scaffold constructs were transplanted into subcutaneous tissue of adult rabbits. MAIN OUTCOME MEASURES:Gross observation and hematoxylin-eosin & Masson staining of neo-cartilage were performed after in vivo culture for 8 weeks. RESULTS:Cell in all groups had a fine adhesion to the scaffold. In both co-culture and chondrocyte groups, the cell-scaffold constructs could maintain the original size and shape during in vivo culture and formed homogenous mature cartilage after 8 weeks of in vivo culture. Furthermore, the neo-cartilages in both groups were similar to each other in gross appearance and histological features. In the bone marrow-derived MSCs group, connective tissue rather than cartilage was found during in vivo culture. CONCLUSION:Chondrocytes can provide a chondrogenic microenvironment to induce a chondrogenic differentiation of bone marrow-derived MSCs and thus promote the chondrogenesis of bone marrow-derived MSCs in vivo.

Objective To assess the abstracts on randomized controlled trials ( RCT) in non-small cell lung cancer ( NSCLC) published in Chinese and their influencing factors.Methods RCT in NSCLC published in Chinese were included according to the CONSORT statement and their influencing factors were analyzed by RevMan 5.3 soft-ware.Results The titles were identified as random, randomization, blinding, statistical method, recruited partici-pants, trial registry and fund-supported, respectively, in 20%of the 2677 abstracts included in this study.Con-clusion The titles are identified as random, randomization, blinding, statistical method, recruited participants, trial registry and fund-supported in RCT published in Chinese.Although the abstracts are improved after the publication of CONSORT, they need to be further brushed up.

Objective To evaluate the biomechanical effectiveness of hyaluronidase (HAase) on rabbit auricular cartilage in the early stage.Methods HAase in 3 different concentrations (75 U/ml,150 U/ml,300 U/ml) were injected subcutaneously to rabbit auricle in 3 groups,while normal saline were injected as control.For the comparison of biomechanism properties among different groups and different time,cartilages were harvested at 3rd and 7th day after injection,followed by stress-relaxation assay.Results Auricular cartilage displayed different levels in control group compared with other groups in elastic modulus (P＜0.05) and maxmum stress (P＜0.05) in 3th day as well as 7th day.While both in the same concentration group,there were also differences between the 3th and 7th day (P＜0.05).Conclusions HAase injection can cause changes in biomechanical properties of auricular cartilage.And 7 days would not be enough for the tissue recovery biomechanically.

Objective To optimize the method of the directional differentiation of adipose-derived stem cells into keratinocytes.Methods Adipose-derived stem cells (ADSCs),separated by collagenase digestion method,were isolated and cultured.Then the expression of surface specific markers CD34,CD44 and CD90 were detected by flow cytometer.The effect of different induced mediums cultured for two weeks on the differentiation of ADSCs into KCs was demonstrated:Group 1,the DMEM supplied with 2％ FBS and 49％ supertant of KCs;group 2,KSFM medium;group 3,DMEM medium supplied with 10％ FBS and 5 μM ATRA;10％ FBS DMEM as the control group.Immunofluorescene staining was applied to detect the expression of keratin CK14 and F-actin.Results A flattened fibroblast-like morphology was observed in cells,the positive expression rate of CD34 was 0.08％,while those of CD44 and CD90 were 99％.The cells that could differentiate into osteoblasts and chondrocytes,indicated that the cells were ADSCs.There was no significant change in the cell morphology in the group 1 under the induction medium;about 10％ of the cells in group 2 were altered;the morphological changes were obvious in group 3,and approximately 20％ of the cells showed irregular polygon.The immunofluorescene staining of the cells in group 3 indicated that the cells showed cobblestone-like phenotype and an organized cytoskeletal network with dense actin fibers at the edges;some cells were positive for CK14.Conclusions ADSCs show higher induction rate under ATRA stimulation.

Nerve growth factor (NGF) can promote the regeneration of peripheral nerve as well as contraction and reepithelization of wound. We constructed a bioengineered dermis containing microencapsulated NGF-expressing NIH-3T3 cells and study the effect of the microencapsule to the bioengineered dermis and seed cells. NGF gene was transfected into NIH-3T3 cells and enclosed into alginate-poly-L-lysine-alginate (APA) microencapsulation and cultivated in vitro. Content of NGF in microencapsules culturing supernatant was measured by enzyme linked immunosorbent assay (ELISA) method. These microencapsules were co-cultured with epidermic cells and fibroblast cells. Bioengineered dermis was constructed with NGF-expressing micorencapsules as seed cells using tissue engineering method. NIH-3T3 microencapsules, empty microencapsules, normal culture media were control groups. After one week culture, the characteristics of the dermis were described by MTT test, the content of hydroxyproline (HP), HE staining and ultrastructure photograph. We found the NGF-expressing microencapsulates can secret NGF steadly after cultured 8w in vitro, promot the proliferation of epidermic cells and secret collagen of fibroblast cells. These functions can maintaine in bioengineered dermis. So NGF-expressing NIH-3T3 microencapsulates can promote the quality of bioengineered dermis.
Alginates ; chemistry ; Animals ; Biocompatible Materials ; chemistry ; Cell Proliferation ; Dermis ; cytology ; Gene Expression Regulation ; Mice ; NIH 3T3 Cells ; Nerve Growth Factor ; genetics ; Polylysine ; analogs & derivatives ; chemistry ; Skin Physiological Phenomena ; Tissue Engineering ; methods ; Transfection ; methods

Objective:To evaluate the reliability and validity of the Chinese version of HEMO-FISS-QoL(HF-QoL) questionnaire (HF-QoL-C) in the Chinese population with hemorrhoids.Methods:From November 2021 to November 2022, a self-constructed general information questionnaire, HF-QoL-C, and the 36-item short form health survey (SF-36), Goligher classification, and Giordano severity of hemorrhoid symptom questionnaire (GSQ) were used to conduct a questionnaire survey on 760 hemorrhoid patients in the anorectal department of six hospitals. The data was analyzed for reliability and validity using SPSS 21.0 and AMOS 26.0 software.Results:The Cronbach's α coefficient of HF-QoL-C and its dimension ranged from 0.831 to 0.960, and the split coefficient was 0.832-0.915. Four common factors were extracted through principal component exploratory factor analysis. Confirmatory factor analysis indicated acceptable structural validity( χ2/ df=8.152, RSMEA=0.097, CFI=0.881, IFI=0.881, NFI=0.867). HF-QoL-C was correlated with SF36 and GSQ( r=-0.694, 0.501, both P<0.01). There were differences in the total score and dimensional scores of HF-QoL-C between surgical and drug treated patients, different grades of Goligher classification for hemorrhoidal disease, and different ranges of hemorrhoid prolapse (all P<0.001). No ceiling effect was found in the total score and the scores of each dimension(0.3%-2.0%). There was a floor effect in both psychological function and sexual activity dimensions (16.7%, 35.1%). Conclusion:HF-QoL-C has good reliability and validity, which can be used to measure the quality of life of Chinese hemorrhoid patients.

FRMD6, a member of the 4.1 ezrin-radixin-moesin domain-containing protein family, has been reported to inhibit tumor progression in multiple cancers. Here, we demonstrate the involvement of FRMD6 in lung cancer progression. We find that FRMD6 is overexpressed in lung cancer tissues relative to in normal lung tissues. In addition, the enhanced expression of FRMD6 is associated with poor outcomes in patients with lung squamous cell carcinoma (n = 75, P = 0.0054) and lung adenocarcinoma (n = 94, P = 0.0330). Cell migration and proliferation in vitro and tumor formation in vivo are promoted by FRMD6 but are suppressed by the depletion of FRMD6. Mechanistically, FRMD6 interacts and colocalizes with mTOR and S6K, which are the key molecules of the mTOR signaling pathway. FRMD6 markedly enhances the interaction between mTOR and S6K, subsequently increasing the levels of endogenous pS6K and downstream pS6 in lung cancer cells. Furthermore, knocking out FRMD6 inhibits the activation of the mTOR signaling pathway in Frmd6^-/- gene KO MEFs and mice. Altogether, our results show that FRMD6 contributes to lung cancer progression by activating the mTOR signaling pathway.