BACKGROUND: Bone marrow mesenchymal stem cells (MSCs) have been widely accepted by medical investigators due to their advantages including easy obtaining, minimal invasion, with infinite proliferation and multi-differential potential, and without immunological rejection in the autologous transplantation. OBJECTIVE: The goal of this study is to isolate and purify rat bone marrow MSCs in vitro, so as to observe the effects of different concentrations of serum containing kidney-tonifying and blood-activating Chinese herbs on the in vitro proliferation of rat bone marrow MSCs.DESIGN: A randomized controlled animal experiment.SETTING: Hip Center, First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine.MATERIALS: Forty healthy male SD rats of SPF grade, weighing 170 to 180 g, were provided by the Laboratory Animal Center, Guangzhou University of Traditional Chinese Medicine. The protocol was performed in accordance with ethics guidelines for the use and care of animals. The involved rats were divided into 4 groups by random digit table with 10 rats in each: normal control group, high-, middle-, and low-concentration groups. METHODS: This study was performed at the Laboratory Animal Center, Guangzhou University of Traditional Chinese Medicine between January and March 2005. Rat bone marrow MSCs were isolated and purified by Percoll density gradient centrifugation, and cultured in vitro to establish rat bone marrow MSCs culture system. Rats in the high-, middle-, and low-concentration groups were intragastrically administrated with 4.4, 2.2 and 1.1 g/kg serum containing kidney-tonifying and blood-activating Chinese herbs, which equaled to 20, 10 and 5 times of adult dosage, respectively. Rats in the normal control group were intragastrically administrated with purified water for 1 week. One hour after the last administration, 6 mL blood was taken from abdominal aorta of each rat under the aseptic condition. Then, it was centrifuged at 2 000 r/min for 15 minutes, and meanwhile serum was collected. 10% rat serum containing kidney-tonifying and blood-activating Chinese herbs was added to the medium in the high-, middle-, and low-concentration groups, while 10% fetal bovine serum was added in the normal control group. MAIN OUTCOME MEASURES: ① MSCs growth status; ② MSCs morphology was observed by HE staining and Giemsa's staining; ③ MSCs antigen expression was detected by an immunocytochemical method; ④ Effects of different concentrations of serum containing kidney-tonifying and blood-activating Chinese herbs on MSCs growth.RESULTS: ①The primarily cultured bone marrow MSCs began to adhere to the wall 24 hours later and 80% of them reached the confluence 7 days later. ② MSCs took appearance in long shuttle shape or polygon. These cells were little. Nuclei were located in the middle part of cells or a little deviation. The ratio of nucleus to cytoplasm was a little high. ③CD44 expression was found in the cytoplasm of mononuclear cells, and colored blue. Partial MSCs expressed c-Kit. Their cytomorphology and phenotypic expression have the characteristics of MSCs. ④Three days after serum containing kidney-tonifying and blood-activating Chinese herbal medicine being added to high-, middle-, and low-concentration groups, the number of bone marrow MSCs was dose-dependently increased as compared with that in the normal control group. CONCLUSION: Serum containing kidney-tonifying and blood-activating Chinese herbs promotes the in vitro proliferation of bone marrow MSCs.

Objective To observe the effect of glucocorticoid on osteoblast-like cell apoptosis and the cell cycle. Methods Osteoblast-like cells OS-732 at the density of 1?105/mL were cultured in vitro with dexamethasone (DMX) at the concentrations of 10-7,10-8 and 10-9 mmol/L. The apoptosis of OS-732 was detected by flow cytometry after the culturing for 24 and 48 hours,and then DNA content was detected and the cell cycle was analysed. Results At the time point of the 24th hour,DNA content at G2/M phase of OS-732 cell cycle was increased obviously in DMX1(10-7 mmol/L)group (P

Objective To study the effects of matrine (MAT) on the apoptosis and the expression of PEG10 in human hepatocarcinoma cell line HepG2. Methods MTT assay was used to determine the proliferation-inhibition activity by MAT to HepG2 cell. JC-1 staining was prepared to detect the change of mitochondrial membrane potential in HepG2 cells after MAT was given. RT-PCR and immunocytochemical method for detecting the PEG10 gene and protein expression levels were used. Results MAT could inhibit the HepG2 cell proliferation above the concentration of 0.125 mg/mL (different from above-->MAT ≥ 0.1 g/L) and in a concentration-dependent and time-dependent manner(P<0.01). JC-1 staining and flow cytometry detection showed that MAT can significantly decrease the mitochondrial membrane potential of HepG2 cells (P < 0.01). The RT-PCR and immunocytochemical staining results showed that 0.5 and 2.0 mg/ml (different from Chinese) MAT could reduce PEG10 gene and protein expression obviously. Conclusion MAT could decrease the expression level of PEG10 gene and inhibited cell proliferation,change the mitochondrial membrane potential and induce HepG2 cell apoptosis.

Objective:To explore the torque effect of controlling root auxiliary arch on the straight wire appliance on maxillary anterior teeth.Methods:22 patients with class Ⅱ1 extraction treatment,treated with straightwire appliance,at the detailing treatment stage were treated with controlling root auxiliary arch over the erect or introverted maxillary incisors for positive torque.The main arch wire of 0.46 mm ×0.64 mm stainless steel edge wire was used in 14 cases,the main arch wire of 0.46 mm stainless steel round wire was used in 8 cases.Cephalometric results before and after maxillary incisor torque were analyzed by t test.Results:After maxillary incisor torque in the 22 cases 1-SN and UIa-NA value increased(P ＜0.01),positive torque was achieved by controlling root auxiliary arch on the upper incisors;the round wire maxillary incisor torque worked faster than the edge wire(P ＜0.05).Conclusion:At the detailing stage of treatment straight wire appliance on front teeth,controlling root auxiliary arch is simple and effective in the treatment of exces-sive erect or lingual inclination of maxillary incisor.

The Forkhead box O1 (FoxO1) transcription factor governs muscle growth, metabolism and cell differentiation. However, its role in myoblast differentiation is unclear. To study the biological function of FoxO1 during differentiation in porcine primary myoblast, we constructed stably FoxO1 over-expressed porcine myoblast mediated by liposome and adopted morphological observation, quantitative real-time RT-PCR and Western blotting methods to analyze FoxO1 and early and late myogenic regulation factors MyoD and myogenin expression. During differentiation the mRNA level of FoxO1 was significantly increased. However, the total protein did not change but the phosphorylation of FoxO1 was upregulated. Furthermore, overexpression of FoxO1 in porcine myoblast decreased MyoD and myogenin mRNA, whereas MyoD protein changed little and myogenin was significantly suppressed (P < 0.05). These results indicated that FoxO1 delays and negatively regulates the porcine myoblast differentiation. Moreover, FoxO1 may play a critical role in muscle fiber-type specification through the inhibition of myogenic regulation factors.
Animals ; Animals, Newborn ; Cell Differentiation ; genetics ; Cells, Cultured ; Forkhead Transcription Factors ; biosynthesis ; genetics ; Muscle, Skeletal ; cytology ; metabolism ; Myoblasts ; cytology ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Swine

BACKGROUND:Nowadays,posterior cruciate ligament injury caused by a sports injury or vehicle injury is more common than people think.Posterior cruciate ligament reconstruction is one of the main treatment methods,but there are still a lot of controversies about the surgical method and ligament selection of posterior cruciate ligament reconstruction. OBJECTIVE:To comprehensively analyze the global application trend of posterior cruciate ligament reconstruction and identify promising research hotspots of posterior cruciate ligament reconstruction based on bibliometrics and visual analysis. METHODS:Publications(articles and reviews)related to posterior cruciate ligament reconstruction from 2000 to 2022 were retrieved from the Web of Science(WOS).The country,institution,publication year,author,journal,average citations per item,H index,title,keywords of publication,and the top 25 cited articles were extracted and analyzed in detail.The VOSviewer/citespace/Pajek software was used to analyze the co-occurrence result of keywords to predict the hotspots of posterior cruciate ligament reconstruction. RESULTS AND CONCLUSION:A total of 664 articles were included.(1)In the past 22 years,the number of posterior cruciate ligament reconstruction articles has shown an increasing trend in general.The top 3 countries(the USA,China,and South Korea)accounted for 65.51%of all articles published.The USA has the largest number of publications.The University of Pittsburgh is the largest contributor.Knee Surgery Sports Traumatol Arthrosc and American Journal of Sports Medicine are the most influential journals.Laprade,Robert F.is the professor who has published the most articles in the field of posterior cruciate ligament reconstruction,and Fanelli,GC is the professor who has the highest total chain strength in the field of posterior cruciate ligament reconstruction.(2)The research direction can be divided into the following five clusters:"posterior cruciate ligament anatomical and biomechanical studies","posterior cruciate ligament reconstruction prognosis,outcome,and complications","posterior cruciate ligament reconstruction surgical method and tendon selection","surgical technique",and"posterior cruciate ligament tear combined with multiple ligament injury".(3)It is concluded that in terms of the trend of previous years,an increasing number of articles related to posterior cruciate ligament reconstruction will be published in the future.The USA is a world leader in the field of posterior cruciate ligament reconstruction.China and South Korea presented great potential in this area.Anatomical and biomechanical research of posterior cruciate ligament and posterior cruciate ligament reconstruction methods and the selection of tendons may be the future hotspots in the field of posterior cruciate ligament reconstruction.

Objective:To explore the application of Tongdu Tiaoshen acupuncture in patients with hemiplegia after stroke.Methods:Randomized controlled trial. A total of 70 patients who met the inclusion criteria for post-stroke hemiplegia in our hospital from January 2020 to December 2021 were divided into two groups, with 35 cases in each group, according to the random number table. The control group was given conventional treatment combined with rehabilitation exercise therapy, and the observation group was given Tongdu Tiaoshen acupuncture on the basis of the above, and both groups were treated continuously for 1 month. Compare of the two groups Fugl-Meyer Assessment (FMA), Modified Ashworth Scale (MAS), National Institutes of Health Stroke Scale (NIHSS), Activity of Daily Living (ADL) score; Brunnstrom stage method was used to assess the patient's motor ability, and Holden walking function grade was used to assess the patient's walking ability. The activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time(TT), and serum fibrinogen (FIB), D-dimer (D-D), plasminogen activator inhibitor 1 (PAI-1) levels were observed by ELISA. Adverse events were recorded and clinical efficacy was evaluated.Results:The total effective rates of the observation group and the control group were 97.14% (34/35) and 77.14% (27/35), respectively, and the difference between the two groups was statistically significant ( χ2=6.25, P=0.012). After treatment, the scores of FMA and ADL in the observation group were significantly higher than those in the control group ( t values were 9.23 and 9.54, respectively, P<0.01), the MAS and NIHSS scores were significantly lower than those in the control group ( t values were 10.23 and 11.97, respectively, P<0.01). After treatment, the distribution of patients in Brunnstrom stage Ⅴ and Ⅵand Holden functional walking stage Ⅳ and Ⅴ in the observation group was significantly higher than those in the control group ( χ2 values were 11.96 and 11.27, respectively, P<0.05). After treatment, APTT, PT and TT in the observation group were significantly longer than those in the control group ( t values were 10.37, 13.57 and 6.54, respectively, P<0.01); serum FIB, D-D and PAI-1 levels were significantly lower than those in the control group ( t values were 12.85, 11.94 and 27.39, respectively, P<0.01). No adverse reactions occurred in both groups. Conclusion:The treatment of post-stroke hemiplegia with Tongdu Tiaoshen acupuncture can improve the effect of clinical treatment and the ability of daily life, neurological function, limb function, movement and walking ability, and stabilize the clotting state.

ObjectiveTo investigate the role and mechanism of Panax notoginseng saponins （PNS） in inhibiting transforming growth factor-β₁ （TGF-β₁）-induced renal tubular epithelial cell injury. MethodNRK-52E renal tubular epithelial cells were cultured and divided into control group， TGF-β₁ group，TGF-β₁+12.5 mg·L^-1 PNS group，TGF-β₁+25 mg·L^-1 PNS group and TGF-β₁+50 mg·L^-1 PNS group. After 48 hours of PNS intervention， the cells and the supernatant were collected， and cell morphology was observed by inverted microscope. Western blot was used to detect the expression of epithelial-mesenchymal transition （EMT）-related proteins and autophagy-related proteins. Flow liquid chromatography for multiple protein quantification and flow cytometry were employed to determine the content of inflammatory factors and apoptosis rate， respectively. ResultCompared with the conditions in the control group， after TGF-β₁ induction， the cells showed a spindle-shaped change and the expression of E-cadherin was down-regulated （P<0.05）， while the expression of α-smooth muscle actin （α-SMA） was up-regulated （P<0.05）. After PNS treatment， most of the cells tended to be normal and reversed the occurrence of EMT. In addition， compared with the conditions in the control group， the level of TNF-α was increased while that of IL-10 was decreased， with elevated apoptosis rate （P<0.05） in the TGF-β₁ group. After PNS treatment， the level of TNF-α was lowered while that of IL-10 was boosted with the increase of the dose， with reduced apoptosis rate （P<0.05）. Moreover， after TGF-β₁ induction， the expression of autophagy-related proteins Beclin 1 and LC3Ⅱ/Ⅰ in renal tubular epithelial cells were up-regulated， while PNS inhibited their expression（P<0.05，P<0.01）. ConclusionPNS had a protective effect on TGF-β₁-induced renal tubular epithelial cells， and the mechanism might be that it reduced inflammation and apoptosis by inhibiting autophagy， thus alleviating TGF-β₁-induced injury.