Objectives To investigate the effects of matrine on the proliferation and apoptosis of SK-NEP-1 cells in vitro, and its possible mechanism. Methods Trials were divided into following groups:control group, 0.5, 1.0 and 1.5mg/ml of ma-trine intervention groups. The inhibition rate of SK-NEP-1 cells treated with different concentration of Matrine was detected by MTT colorimetric assay. Apoptosis rate was detected by flow cytometry (FCM). RT-PCR analysis was employed to measure the PDCD4 mRNA expression. Results Matrine (final concentrations=0.5, 1.0 and 1.5mg/ml) could induce apoptosis and inhibit the growth of SK-NEP-1 cells. Compared with the controls without matrine treatment (8.81±3.71)%, the inhibition rates of SK-NEP-1 cells were (20.79 ± 6.20)%, (31.25 ± 5.07)%, and (51.15 ± 12.70)%, respectively;the apoptotic rates of SK-NEP-1 cells treated with different concentration of matrine were (13.67±0.78)%,(17.43±1.65)%and (20.80±1.54)%, respectively. Significant difference in the inhibition and apoptotic rates of SK-NEP-1 cells between each drug group and control group was observed(P<0.05), and the inhibition and apoptotic rates of SK-NEP-1 cells increased gradually with increased matrine concentration, thus exhibiting a dose-dependent effect(P<0.05). To the expression of CXCR4 mRNA,the grey levels of SK-NEP-1 cells treated with matrine intervention group (final concentrations=0.5, 1.0 and 1.5 mg/ml) were (0.720 ± 0.058), (0.540 ± 0.095) and (0.307 ± 0.050), respectively. The mRNA expression of CXCR4 was seen in SK-NEP-1 cells. Compared with control group, the expres-sion of CXCR4 mRNA was decreased significantly in matrine intervention group (P<0.01).There were significant difference in CXCR4 mRNA level among the SK-NEP-1 cells treated with 0.5,1.0,1.5mg/mL of matrine (P<0.01). Conclusions Matrine could induce apoptosis and inhibit the growth of SK-NEP-1 cells in a dose-dependent way which may be associated with the down-regulated CXCR4 expression in SK-NEP-1 cells.

Objective To investigate the effect of Matrine on the expression of X-linked inhibitor of apoptosis protein (XIAP) in human rhabdomyosarcoma RD cells in vitro.Methods Cultured human rbabdomyosarcoma RD cells were divided into Matrine intervention groups (0.5 g/L,1.0 g/L and 1.5 g/L) and a control group.The proliferation-inhibition rates in RD cells treated with different concentrations of matrine were detected by methylthiazolyl blue colorimetric assay.Flow cytometry analysis was performed for the apoptosis rates of RD cells.Reverse transcription-polymerase chain reaction analysis was used to measure the XIAP mRNA expression.Results There was a significant difference in the proliferation-inhibition rates [0.5 g/L Matrine group:(15.84 ± 2.58)％,1.0 g/L Matrine group:(23.13 ±4.19)％,1.5 g/L Matrine group:(30.32 ±3.02)％,and the control group:(8.92 ± 1.23)％];apoptotic rates [0.5 g/L Matrine group:(12.33 ± 1.15)％,1.0 g/L Matrine group:(16.67 ± 0.99)％,1.5 g/L Matrine group:(25.33 ± 1.91)％,and the control group:(9.47 ± 0.96)％];XIAP mRNA expression(0.5 g/L Matrine group:0.633 ± 0.046,1.0 g/L Matrine group:0.441 ± 0.055,1.5 g/L Matrine group:0.326 ± 0.065,control group:0.794 ±0.029)in RD cells among 0.5 g/L,1.0 g/L,1.5 g/L Matrine groups and the control group (F =14.15,83.37,50.57,all P ＜ 0.05).The proliferation-inhibition and apoptotic rates in RD cells were gradually increased with the increasing Matrine concentration.The expression of XIAP mRNA was significantly decreased in different Matrine groups compared with the control group,exhibiting a dose-dependent manner.Conclusions Matrine can inhibit the proliferation of RD cells and induce the apoptosis in a dose-dependent manner,which may be related to the down-regulated XIAP mRNA.

Objective To investigate the effects of matrine on proliferation and apoptosis of human rhabdomyosarcoma(RMS) RD cells line in vitro,to study the regulatory mechanism of Wnt/β-catenin pathway-influenced apoptosis of RD cells line by detecting the expressions of β-catenin protein,Bcl-2 protein and caspase-3 protein,and to explore Wnt/β-catenin mechanism during the process of RMS.Methods The human RMS RD cells line was treated with matrine of different concentrations (0.5,1.0,1.5,2.0 g/L)for 48 hours respectively,and the proliferation inhibition rates of different concentrations of matrine on RD cells were detected by methyl thiazolyl tetrazolium assay,while the apoptosis rates by flow cytometry (FCM) and the expressions of β-catenin,Bcl-2 and caspase-3 by Western blot.Results The proliferation inhibition rates between control group and different concentrations of matrine groups were(13.70 ±0.25)％,(33.16 ±0.11)％,(42.96 ±0.90)％,(56.26 ±0.79)％ and (67.89 ±0.63)％,respectively.The apoptosis rates were (5.49 ± 0.96) ％,(17.23 ± 5.03) ％,(25.84 ± 4.17) ％,(36.08 ± 3.68) ％and (47.79 ± 4.82) ％,respectively.The highest expression of β-catenin and Bcl-2 proteins and the minimum amount of caspase-3 protein were found in the control group.After intervention of matrine,the expressions of β-catenin and Bcl-2 reduced while the amount of caspase-3 rose significantly,which was concentration-dependent obviously.Differences were found between every concentration of matrine group with control group according to statistics (all P ＜ 0.05).Conclusions Matrine can inhibit proliferation and induce apoptosis of RD cells.Matrine can down-regulate the expression of β-catenin and Bcl-2 proteins in RD cells,while the amount of caspase-3 protein rises.Wnt/β-catenin signal pathway plays an important role in the apoptosis of RD cell induced by matrine,and its downstream proteins Bcl-2 and caspase-3 are also involved in the regulation of this process.

Objective The purpose of this paper is to study the relationship between blood concentration and brain tissue drug concentration by different dose of TMX chemotherapy acute lymphoblastic leukemia in mice. Methods 4 weeks, health Kun Ming mice 80: establishment acute lymphoblastic leukemia mice model,20 mice were randomly selected to take the femur bone marrow biopsy bone marrow OK for model verification; the remaining 60 acute lymphoblastic leukemia mice were allocated randomly 6 groups of 10 mice in each group, respectively A, B, C, D, E, F groups. And collected blood 0.5 ml and brain tissue 0.4 g individually at 0.5 hour in every group. We used supernatant of centrifugation blood and brain homogenate to detected drug concentration by fluorescence polarization immunoassay. Results The mean blood concentration of MTX of six groups A, B, C, D, E, F are (39.08±5.18) μmol/L, (15.86±1.02)μmol/L, (8.67± 5.43)μmol/L, (68.29±5.19)μmol/L, (29.55±6.22)μmol/L, (13.98±1.12)μmol/L, respectively. Compared the mean blood concentration of MTX of each group there are statistical significance (P<0.05). The mean concentration of MTX of six groups in brain tissue are followed by A group (1.05±0.26)μmol/L, B group (0.61±0.25)μmol/L, C group (0.48±0.25)μmol/L, D group (2.07±0.35)μmol/L, E group (1.27±0.21)μmol/L, F group (0.59±0.69)μmol/L. Compared the mean concentration of MTX of each group in brain tissue there are statistical significance (P<0.05). MTX concentration in blood and in brain tissue of correlation coefficient followed by 0.82, 0.75, 0.19, 0.81, 0.55, 0.43. Conclusion The chemotherapy acute lymphoblastic leukemia mice of HDMTX scheme, the peak of blood concentration and brain tissue drug concentration is come after injected MTX 0.5 hour, MTX 5 g/m~2 is better permeation blood-brain barrier and more easy make brain tissue drug concentration to reach effectively therapeutic concentration than MTX 3 g/m~2.

Pig copper zinc superoxide dismutase (CuZnSOD) is an important antioxidant enzyme. Some studies focused on the function of CuZnSOD gene, but the transcriptional regulation of the CuZnSOD gene is not yet fully elucidated. Therefore, the aims of the study were to determine the core promoter region and to explore its mechanism of transcriptional regulation. The 853 bp DNA sequence of 5'-flanking promoter was amplified by performing PCR. A series of CuZnSOD promoter fragments with gradually truncated 5'-end were produced by nested PCR and inserted into pGL3-Basic vector. The activities of the promoters were measured by the dual-luciferase assay system after transient transfection into the NIH/3T3 cells. The results demonstrated that there were 2 potential transcription start sites in the regions from initiation codon to -87 bp and -266 bp, respectively. The region from -383 bp to +67 bp in CuZnSOD gene promoter showed higher activity than other regions, and further deletion analysis demonstrated that the region from -75 bp to -32 bp contained an essential promoter sequence for pig CuZnSOD gene transcription. In addition, several potential transcription factor binding sites were predicted with bioinformatics method. These results suggest that these transcription factor binding sites may be involved in the transcriptional regulation of CuZnSOD gene.
Animals ; Cloning, Molecular ; Gene Expression Regulation ; Genetic Vectors ; Mice ; NIH 3T3 Cells ; Promoter Regions, Genetic ; Superoxide Dismutase ; genetics ; Swine ; Transfection

Objective The aim of this study was to investigate the relationship between phosphotyrosine interac-tion domain containing 1 ( PID1 ) gene and variation in intramuscular fat ( IMF ) content and the possibility to generate transgenic animals by testicular injection .Methods Expression vector pIRES2-acGFP-PID1 carrying pig PID1 gene was incubated with transfection reagents and injected into the testes of male New Zealand rabbits .We examined the F1 genera-tion by fluorescence detection , PCR, Western blotting and measuring the IMF content .The F1 generation gave birth to the F2 generation.Then we examined the F2 generation through detecting the positive rate and the IMF content .Results The exogenous PID1 gene and fluorescent protein gene were expressed at different levels both in the F 1 generation and the F2 generation, and the positive rates were 35.88%and 34.33%, respectively.The IMF content was significantly elevated (P<0.05) in the transgenic positive individuals compared with the negative ones and the control group , and the PID1 protein expression was similarly higher .Conclusions The results of this study demonstrate that PID 1 gene affects intramuscular fat content significantly .Moreover, the results of our analysis provide further evidence that transgenic animals can be gener -ated by testicular injection , and the exogenous gene can be inherited steadily .

With the development of molecular biology research, targeted therapy has become the fourth effective method for the treatment of malignant tumors after surgery, radiotherapy, and chemotherapy. Unlike the traditional chemotherapeutic drugs, molecular targeted drugs possess the advantages of high specificity, definite curative effects, and less adverse effects. CLDN18.2 protein is a transmembrane protein that is highly selective and stably overexpressed during the development of various malignant tumors, especially gastrointestinal tumors, and metastases. A specific antibody against this protein, claudiximab (zolbetuximab/IMAB362), has achieved great success in recent clinical trials. Thus, CLDN18.2 protein is expected to be a safe and effective molecular target for targeted therapy in some malignancies.

Objective To analyze the clinical effects of arthroscopic en masse repair with footprint ending shift using double-row suture-bridge technique for delaminated rotator cuff tears under tension.Methods A total of 58 patients with delaminated rotator cuff tears under tension from August 2013 to August 2016 who underwent arthroscopic en masse repair using doublerow suture-bridge technique were retrospectively analyzed.There were 33 males and 25 females with a mean age of 53.0±7.8 years (range 39-74) with 24 patients left side involved and 34 right side.They were divided into 2 groups to receive en masse repair either footprint ending shift or on the footprint.There were 28 patients with footprint ending shift and 30 patients on the footprint.Clinical effects were evaluated by University of California Los Angeles (UCLA) score,American Shoulder and Elbow Surgeons (ASES) score,visual analogue scale (VAS),Constant-Murley score and shoulder range of motion at preoperatively and postoperatively.Results The average follow-up duration was 23.2±0.8 months (range 21-24).The two groups were compatible with no significant difference in age,gender,tear size,follow-up duration,preoperative function and range of motion of the shoulder joint (P＞0.05).At the last follow up,the UCLA,ASES,VAS,Constant-Murley scores and shoulder range of motion in the group footprint ending shift were respectively 32.4±2.5,12.8±0.9,1.0±1.1,93.4±5.6,158.3°±9.3°,58.9°±5.0° with significantly differences compared with preoperative scores (P ＜ 0.05).The postoperative value in the group on footprint were respectively 31.6±2.9,12.8±0.9,0.7 ± 1.2,91.3±7.1,156.1°± 10.7°,59.6°±4.6° with significantly differences compared with the preoperative scores (P ＜ 0.05).There were no significant difference between the two groups (P ＞ 0.05).The operation duration in the group footprint ending shift was 100.9±6.0 min,while that in the group on footprint was 106.6±6.1 min.There was significantly difference in the operation duration between two groups (t=-3.600,P=0.001).Conclusion Arthroscopic en masse repair using double-row suture-bridge technique can successfully treat delaminated rotator cuff tears under tension.Compared with arthroscopic en masse repair on footprint using double-row suture-bridge technique,the footprint ending shift is easier and time saving without significant difference in function of the shoulder joint and the range of motion in repair of delaminated rotator cuff tear under tension.

Objective To evaluate the clinical efficacy of arthroscopic bursal layer-only double-row suture-bridge repair for delaminated rotator cuff tear which is difficult to reposit in comparison with separate double-layer repair and whole-layer repair.Methods From May 2013 through June 2016,82 patients with delaminated rotator cuff tear difficult to reposit were treated at Department of Joint Surgery,The Affiliate Hospital to Chengde Medical University.They were 47 males and 35 females with a mean age of 53.0 ± 7.9 years.They were divided into 3 groups according to their surgical procedures.In group A,28 cases were treated by arthroscopic whole-layer double-row suture-bridge procedure;in group B,29 cases were treated by arthroscopic separate double-layer double-row suture-bridge procedure;in group C,25 cases were treated by arthroscopic bursal layer-only double-row suture-bridge procedure.The 3 groups were compared in terms of University of California Los Angeles (UCLA) score,American Shoulder and Elbow Surgeons (ASES) score,visual analogue scale (VAS),Constant shoulder score,range of motion of shoulder joint and rotator cuff retear preoperatively and postoperatively.Results The patients in the 3 groups were comparable because their preoperative general data showed no significant significances (P ＞ 0.05).The operation time for groups A,B and C was respectively 105.5 ±5.6 min,117.4 ±6.9 min and 88.0 ±4.2 min,showing significant differences between the 3 groups (P ＜ 0.05).The 82 patients were followed up for 21 to 24 months (average,23.3 months).At 24 months postoperatively,UCLA,ASES,VAS,Constant score,shoulder anteflexion and lateral extorsion were respectively 32.4 ± 2.5,12.8 ± 0.9,1.0 ± 1.1,93.4 ± 5.6,158.3° ± 9.3°and 58.9°±5.0°in group A,32.2±2.5,12.9±1.0,0.9±1.0,92.8±6.0,156.4°±9.5°and 59.3°± 5.6° in groups B,and32.4±2.4,12.9±0.9,0.7±0.9,94.3±5.2,156.0°±9.5°and57.6°°5.4°in group C,showing no significant differences between the 3 groups (P ＞ 0.05).The occurrence of rotator cuff retear in groups A,B and C were respectively 17.9％ (5/28),13.8％ (4/29) and 12.0％ (3/25),showing no significant differences between the 3 groups (P ＞ 0.05).Conclusions In repair of delaminated rotator cuff tear difficult to reposit,although the arthroscopic bursal layer-only double-row suture-bridge repair is similar to conventional arthroscopic whole-layer double-row suture-bridge repair and arthroscopic separate double-layer double-row suture-bridge repair in functional recovery and range of motion of the shoulder and incidence of rotator cuff retear,it can reduce obviously operation time and make the operation easier.

Objective:To compare the effects of patellar position on the balance of soft tissue during the operation and the postoperative outcomes in minimally invasive total knee arthroplasty via a midvastus approach.Methods:From December 2018 to February 2019, a total of 55 patients were enrolled for primary total knee arthroplasty via a midvastus approach. All patients were randomly divided into 2 groups by random number table. During the operation, the gap balance technique was used with patella reduced or subluxated to complete the osteotomy and balance of soft tissue. The changes of gap and varus-valgus angle were compared between the patellar reduction group and the patellar subluxation group in both extension and flexion position. These data were also compared before and after reducing patellar in the patellar subluxation group. Furthermore, the differences of femoral prosthesis rotation, mechanical femoral axis to tibial axis angle, Knee Society score (KSS), visual analogue scale (VAS) and range of motion (ROM) were compared between the two groups.Results:All patients finished 6 months follow-up, including 27 patients in the patellar reduction group and 28 patients in the patellar subluxation group. After osteotomy and soft tissue balance during the operation, there was no significant difference in gap and varus-valgus angle between two groups in either extension or flexion position. While in the patellar subluxation group, the flexion gap was 10.5±0.3 mm with patella subluxated, less than 11.0 ± 0.3 mm after reducing the patella. The varus-valgus angle was 1.5±0.3 with patella subluxated, less than 2.3±0.4 degree after reducing the patella. The difference was statistically significant ( t=4.180, P<0.001; t=7.642, P<0.001). There was no significant difference in extension gap and varus-valgus angle before and after patella reduction in the patellar subluxation group ( P>0.05). The rotation angle of the femoral component in the patellar subluxation group was -0.49°±1.2°, and the external rotation angle was smaller than that in the patellar reduction group (0.24°±1.3°). The difference was statistically significant ( t=2.116, P=0.039). At one month after operation, ROM of the patellar reduction group was 109.6°±8.5° which was higher than that of the patellar subluxation group (104.9°±8.6°, t=2.048, P=0.046). There was no significant difference in ROM between the two groups at 3 and 6 months ( P>0.05). Moreover, there was no significant difference in KSS and VAS at 1, 3 and 6 months after operation ( P>0.05). Conclusion:In the minimally invasive total knee arthroplasty, it is suggested to balance the soft tissue as much as possible with the patellar reduced. Otherwise, the consequence of increased flexion space, increased varus and increased internal rotation of femoral prosthesis should be considered. The patients undergoing soft tissue balance with patella reduced have better ROM in the early stage postoperatively.