Novel nano-microspheres containing chitosan, hyaluronic acid, and chondroitin sulfate deliver growth and differentiation factor-5 plasmid for osteoarthritis gene therapy.

OBJECTIVE: To construct a novel non-viral vector loaded with growth and differentiation factor-5 (GDF-5) plasmid using chitosan, hyaluronic acid, and chondroitin sulfate for osteoarthritis (OA) gene therapy. METHODS: Nano-microspheres (NMPs) were prepared by mixing chitosan, hyaluronic acid, and chondroitin sulfate. GDF-5 plasmid was encapsulated in the NMPs through electrostatic adsorption. The basic characteristics of the NMPs were observed, and then they were co-cultured with chondrocytes to observe their effects on extracellular matrix (ECM) protein expression. Finally, NMPs loaded with GDF-5 were injected into the articular cavities of rabbits to observe their therapeutic effects on OA in vivo. RESULTS: NMPs exhibited good physicochemical properties and low cytotoxicity. Their average diameter was (0.61±0.20) µm, and encapsulation efficiency was (38.19±0.36)%. According to Cell Counting Kit-8 (CCK-8) assay, relative cell viability was 75%-99% when the total weight of NMPs was less than 560 µg. Transfection efficiency was (62.0±2.1)% in a liposome group, and (60.0±1.8)% in the NMP group. There was no significant difference between the two groups (P>0.05). Immunohistochemical staining results suggested that NMPs can successfully transfect chondrocytes and stimulate ECM protein expression in vitro. Compared with the control groups, the NMP group significantly promoted the expression of chondrocyte ECM in vivo (P<0.05), as shown by analysis of the biochemical composition of chondrocyte ECM. When NMPs were injected into OA model rabbits, the expression of ECM proteins in chondrocytes was significantly promoted and the progression of OA was slowed down. CONCLUSIONS: Based on these data, we think that these NMPs with excellent physicochemical and biological properties could be promising non-viral vectors for OA gene therapy.

Construction of human telomerase reverse transcriptase periodontal ligament cell line mediated by adenovirus / 华西口腔医学杂志

OBJECTIVE@#This study aims to establish an effective and stable periodontal ligament cell line stably expressing human telomerase reverse transcriptase (hTERT) gene by using the adenovirus method.@*METHODS@#Polymerase chain reaction (PCR) was used to amplify the full length of hTERT gene to construct recombinant adenovirus plasmid pAd-pshuttle-cmv-hTERT. Packaged adenovirus particles were used for infection of human periodontal ligament cells. The expression levels of hTERT and osteogenic genes, such as alkaline phosphatase, Runt-related transcription factor 2, bone sialoprotein, osteocalcin, osteopontin, and collagen Ⅰ mRNA, were detected by quantitative real-time PCR (qRT-PCR). The ability of osteogenic differentiation was observed by alizarin red staining, and the cell proliferation was determined by CCK-8.@*RESULTS@#Adenovirus particles containing the hTERT gene were successfully constructed and infected with periodontal ligament cells. The infected cells were similar to normal periodontal ligament cells. The qRT-PCR results showed that hTERT and osteogenesis-associated genes were highly expressed in the periodontal ligament cell lines constructed by adenoviruses. Alizarin red staining showed that the periodontal ligament cell line had strong osteogenic differentiation capability. CCK-8 showed that the periodontal ligament cell line had strong proliferation capability.@*CONCLUSIONS@#The human periodontal ligament cell line with high efficiency and stable expression of hTERT was established by the adenovirus method, thereby providing an ideal cell line for studying the mechanism of periodontal regeneration.

Analysis on Outpatient Prescriptions of TCM Decoction Pieces in 2016 in Chongqing Hospital of Traditional Chinese Medicine / 中国中医药信息杂志

Objective To understand the present situation and regularity of the application of TCM prescriptions in Chongqing Hospital of Traditional Chinese Medicine (hereinafter referred to as "our hospital"); To provide references for TCM pharmacy management and rational medication in clinic. Methods 8756 TCM prescriptions in our hospital from January to December in 2016 were randomly selected and the method of subtotals was used to collect statistics. The basic rules and existing problems of the TCM prescriptions were analyzed. Results The TCM prescriptions issued more in the famous doctor's center (27.67%), preventive treatment of disease center (13.67%) and oncology department (13.35%); elderly patients took the lead (45–75 years old accounted for 65.90%); prescriptions with 16–20 Chinese herbs added up to 4326 (49.41%); prescriptions within 3–7 dosages were 4921 in total (56.20%);single prescription cost within 100–300 yuan were 2777 in all (31.71%); prescriptions of single dosage cost 21–80 yuan were 5500 altogether (62.81%). Conclusion The application of TCM prescriptions in our hospital is basically rational, but problems such as too many herbs in one prescription, overdose use of some prescriptions and higher amount still exist.

Comparative anatomical study on subinguinal and ilioinguinal approaches and clinical application in acetabular fracture / 中国骨伤

<p><b>OBJECTIVE</b>To compare anatomical differences between subinguinal and ilioinguinal approaches for the treatment of acetabular fracture and investigate clinical therapeutic effect of subinguinal approach.</p><p><b>METHODS</b>Seven fresh human bodies were chosen, comparative study were performed on the right and left side on the same specimen. Ilioinguinal approaches were adopted on the left and subinguinal were adopted on the right. Inner part of incision started to sun wild above pubic symphysis at 2 cm, and lateral incision ranged from iliac to anterior superior spine about 5 cm. Length and transverse diameter of the first window exposed and lliopsoas freeness were tested and compared. Fifteen acetabular fracture patients treated through subinguinal approach were compared from May 2010 to August 2012. Among all patients, including 12 males and 3 females aged from 20 to 65 years old with an average of 40.6 years old. Matta criteria were used to evaluate clinical outcomes.</p><p><b>RESULTS</b>Length and transverse diameter of the first window exposed and lliopsoas freeness through subinguinal approach were better than through ilioinguinal approach (P<0.01). In 15 patients with acetabular fracture, 10 patients obtained anatomical reduction and 10 patients got satisfied reduction in accordance with Matta criteria. X-ray results of all patients were excellent.</p><p><b>CONCLUSION</b>Compared with ilioinguinal approach, subinguinal approach could enlarge visualization of the first window and simplify surgical procedure. It is an ideal approach to expose anterior and anterior-medialis wall of acetabulum and anterior hip capsule.</p>

Regulation of the self-renewal and differentiation of spermatogonial stem cells / 中华男科学杂志

Spermatogonial stem cells (SSCs) play an important role in spermatogenesis and have a unique mode of replication. A single SSC can produce two differentiating cells, or one stem cell and one differentiating cell. The self-renewal and differentiation of SSCs are precisely regulated as relating the niche of SSCs, glial cell line-derived neurotrophic factor, and several signaling pathways. This article reviews the self-renewal and differentiation of SSCs and their regulation mechanisms, which may offer a deeper insight into spermatogenesis and male infertility and pave a theoretical ground for studying testicular tumorigenesis and searching for new potential approaches to the treatment of testicular cancer and other related diseases.