Clinical analysis of MAP in transabdominal laparoscopic radical nephrectomy of renal cell carcinoma / 现代泌尿外科杂志

【Objective】 To evaluate the significance of Mayo adhesive probability (MAP) in predicting surgical difficulty and postoperative recovery in patients with renal cell carcinoma (RCC) undergoing laparoscopic radical nephrectomy (LRN). 【Methods】 The clinical data of 168 RCC patients who received transabdominal LRN during Jan.2017 and Dec.2020 were retrospectively analyzed. According to MAP, the patients were divided into low MAP group (n=100) and high MAP group (n=68). The differences in perioperative clinical data were compared between the two groups. 【Results】 Compared with low MAP group, the high MAP group had longer operation time (P<0.001), more intraoperative blood loss (P<0.001), higher Clavien-Dindo grade complications (P=0.008), longer hospital stay (P=0.003), higher levels of c-reactive protein (P=0.030) and IL-6 (P=0.009), lower levels of albumin (P<0.001) and prealbumin (P=0.020). 【Conclusion】 MAP can assess the risk of prolonged operation time, increased bleeding during transabdominal LRN, and postoperative recovery, thus guiding the preoperative planning.

Increased UCP1 mRNA expression of perirenal fat predicts a poor prognosis of clear cell renal cell carcinoma / 中华内分泌外科杂志

Objective:To evaluate the effect of uncoupling protein 1 (UCP1) expression of perirenal fat on the prognosis of clear cell renal cell carcinoma (ccRCC) .Methods:From Feb. 2013 to Oct. 2013 and Mar. 2015 to Oct. 2015, 98 patients with ccRCC who underwent retroperitoneal laparoscopic radical nephrectomy were analyzed. UCP1 mRNA of perirenal fat around tumor was detected by RT-qPCR. Preoperative Computed tomography (CT) images were used to evaluate the thickness and adhesiveness of perirenal fat. According to the UCP1 mRNA value, the patients were divided into high UCP1 group (42 cases) and low UCP1 group (56 cases) . The general clinical data, perirenal fat thickness and adhesiveness were compared, and Kaplan Meier curve was used to evaluate the difference of progression free survival (PFS) between the two groups. Univariate and multivariate Cox analysis were used to determine the potential independent prognostic factors of PFS.Results:In the high UCP1 group, the renal fat thickness, the ratio of fat adhesion, the ratio of Ⅲ to Ⅳ in Fuhrman grade and the ratio of >T2 in T stage were higher than those in the low UCP1 group[ (13.84±2.41) vs (10.75±1.99) , 42.86% vs 16.07%, 28.57% vs 8.93%, 21.43% vs. 5.36%; P=0.000, P=0.003, P=0.011, P= 0.037]. During the follow-up period (median, 62.0 months) , 15 cases (12 cases of high UCP1 group, 3 case of low UCP1 group) developed tumor progression. Kaplan Meier curve showed that PFS of high UCP1 group was worse than that of low UCP1 group (71.43% vs 94.64%, P=0.001) . Cox regression analysis showed that high UCP1 expression and high T stage were significantly correlated with low PFS ( β=1.334, RR=3.796, 95% CI=1.009-14.280, P= 0.048; β=2.886, RR=17.930, 95% CI=5.538-58.047, P=0.000) . Conclusions:The increased UCP1 expression of perirenal fat may be an independent risk factor of tumor progression in ccRCC. Combined with the assessment of browning of perirenal adipose tissue may be helpful for risk stratification of ccRCC patients after surgery.

Application effect of hospital‐community‐family ternary linkage management in patients with post‐stroke depression／ / 心血管康复医学杂志

To explore application effect of hospital‐community‐family ternary linkage management in pa‐tients with post‐stroke depression .Methods : A total of 92 patients with post‐stroke depression in our hospital were randomly and equally divided into routine treatment group and linkage management group (receive intervention of hospital‐community‐family ternary linkage management ) , both groups were intervened for three groups .Scores of self‐rating anxiety scale (SAS) ,self‐rating depression scale (SDS) ,36‐Item Short Form Health Survey (SF‐36) and satisfaction for medical service of patients were compared between two groups before and after intervention .Re‐sults : Compared with before treatment , there were significant reductions in scores of SAS and SDS , and significant rise in SF‐36 score and each dimension score of satisfaction for medical service in two groups after three‐month inter‐vention , P＝0.001 all ;compared with routine treatment group after three‐month intervention , there were signifi‐cant reductions in scores of SAS [ (41. 10 ± 1.73) scores vs .(38. 22 ± 1. 53) scores] and SDS [ (43. 14 ± 1.93) scores vs.(39. 44 ± 1.85) scores] , and significant rise in SF‐36 score [ (68.35 ± 4.44) scores vs .(87.64 ± 4.36) scores] and each dimension score of satisfaction for medical service of patients in linkage management group , P＝0.001 all. Conclusion : Hospital‐community‐family ternary linkage management possesses significant intervention effect on pa‐ tients with post‐stroke depression .It can significantly improve anxiety and depression emotion , quality of life and satisfaction for medical care service in these patients , which is worth extending .

Analysis of clinical characteristics and surgical results in patients with chronic subdural hematoma in different ages / 中国医师杂志

Objective@#To investigate the clinical characteristics and surgical results of patients with chronic subdural hematoma (CSDH) in different age groups.@*Methods@#150 CSDH patients admitted to our hospital from January 2014 to January 2019 were selected as the research objects, and their clinical data were reviewed and collected. According to different age , they were divided into three groups: the <55 year old group (n=46), the 55-70 year old group (n=62), and the >70 year old group (n=42). The imaging examination results and clinical manifestations of three groups of patients in different age groups were compared. Combined with patients' wishes and surgical indications, 150 patients were treated with traditional burr hole and drainage and modified operation. The results of different operation methods in three groups were compared.@*Results@#The incidence of headache/dizziness in the 55-70 year old group was higher than that in >70 year old group (P<0.05). The incidence of language disorder in the 55-70 year old group and >70 year old group was higher than that in <55 year old group , with statistically significant difference (P<0.05). The incidence of limb paralysis in the 55-70 year old group and the >70 year old group was higher than that in <55 years old group, with statistically significant difference (P<0.05). The incidence of mental disorders in the >70 year old group was higher than that in 55-70 year old group and <55 year old group, with statistically significant difference (P<0.05). The incidence of fecal incontinence in the >70 year old group was higher than that in 55-70 year old group and <55 years old group, with statistically significant difference (P<0.05). The imaging examination results of the three groups showed significant differences (P<0.05). According to the estimation of the amount of hematoma by the Dutian (DOTA) formula, the amount of hematoma in the group of >70 years old was significantly higher than that in the group of <55 years old and 55-70 years old, with statistically significant difference (P<0.05). Among the three groups, the proportion of patients with good prognosis after modified operation was higher than that of traditional operation, and the recurrence rate was lower than that of traditional operation, with statistically significant difference (P<0.05). The proportion of good prognosis and recurrence rate in group >70 years old were lower than those in the group of <55 years old and 55-70 years old (P<0.05).@*Conclusions@#CSDH patients of different ages have different characteristics in imaging examination results and clinical manifestations. Traditional burr hole drainage and modified operation can achieve certain results, but modified operation is more effective and can reduce the incidence of postoperative complications, which is worth popularizing.

Analysis of clinical characteristics and surgical results in patients with chronic subdural hematoma in different ages / 中国医师杂志

Objective To investigate the clinical characteristics and surgical results of patients with chronic subdural hematoma (CSDH) in different age groups.Methods 150 CSDH patients admitted to our hospital from January 2014 to January 2019 were selected as the research objects,and their clinical data were reviewed and collected.According to different age,they were divided into three groups:the ＜ 55 year old group (n =46),the 55-70 year old group (n =62),and the ＞ 70 year old group (n =42).The imaging examination results and clinical manifestations of three groups of patients in different age groups were compared.Combined with patients'wishes and surgical indications,150 patients were treated with traditional burr hole and drainage and modified operation.The results of different operation methods in three groups were compared.Results The incidence of headache/dizziness in the 55-70 year old group was higher than that in ＞ 70 year old group (P ＜ 0.05).The incidence of language disorder in the 55-70 year old group and ＞ 70 year old group was higher than that in ＜ 55 year old group,with statistically significant difference (P ＜0.05).The incidence of limb paralysis in the 55-70 year old group and the ＞70 year old group was higher than that in ＜ 55 years old group,with statistically significant difference (P ＜ 0.05).The incidence of mental disorders in the ＞ 70 year old group was higher than that in 55-70 year old group and ＜ 55 year old group,with statistically significant difference (P ＜ 0.05).The incidence of fecal incontinence in the ＞ 70 year old group was higher than that in 55-70 year old group and ＜ 55 years old group,with statistically significant difference (P ＜ 0.05).The imaging examination results of the three groups showed significant differences (P ＜ 0.05).According to the estimation of the amount of hematoma by the Dutian (DOTA) formula,the amount of hematoma in the group of ＞70 years old was significantly higher than that in the group of ＜ 55 years old and 55-70 years old,with statistically significant difference (P ＜0.05).Among the three groups,the proportion of patients with good prognosis after modified operation was higher than that of traditional operation,and the recurrence rate was lower than that of traditional operation,with statistically significant difference (P ＜ 0.05).The proportion of good prognosis and recurrence rate in group ＞ 70 years old were lower than those in the group of ＜ 55 years old and 55-70 years old (P ＜0.05).Conclusions CSDH patients of different ages have different characteristics in imaging examination results and clinical manifestations.Traditional burr hole drainage and modified operation can achieve certain results,but modified operation is more effective and can reduce the incidence of postoperative complications,which is worth popularizing.

Mechanism Underlying Antitumor Effects of Sinomenine / 中国结合医学杂志

Sinomenine (SIN) is a bioactive alkaloid compound extracted from a Chinese medicinal plant Sinomenium acutum. It is a multitarget antitumor natural substance. Various mechanisms have been proposed for the antitumor effects of SIN, such as direct cytotoxicity, induction of apoptosis, sensitization attenuating radiotherapy and chemotherapy, reversal of drug resistance, resistance to distant metastasis, and antiangiogenesis. SIN can be used as a tumor cell killer and an adjuvant to radiotherapy and chemotherapy. However, recent studies are mostly limited to the basic experimental stage; no systematic clinical studies have yet been reported. Therefore, this paper aimed to review the mechanism underlying the antitumor effects of SIN by consulting relevant domestic and foreign studies and to provide a relevant reference for further development, use, and exploration of SIN.

Priliminary study on efficacy and safety of umbilical cord mesenchymal stem cells infusion for treating refractory rheumatoid arthritis / 重庆医学

Objective To explore the efficacy and safety of umbilical cord mesenchymal stem cells(UC-MSCs) infusion in treating refractory rheumatoid arthritis(RA).Methods Four cases of refractory RA and invalid by improving disease condition and anti-rheumatic drugs in this hospital from December 2014 to November 2015 underwent once UC-MSCs suspension intravenous infusion after signing the informed consent.The cells number was 1 × 106 per kg.We evaluated the tender and swollen joints number,erythrocyte sedimentation rate(ESR),C-reactive protein(CRP),28-joint disease activity score(DAS28),visual analog scale(VAS)pain score and related safety indices were compared between before and after infusion.Results After infusion treatment in 3 cases,the tender and swollen joints number,ESR and DAS28(4) score were significantly improved,but another case was ineffective.No anaphylaxis,rejection reaction or adverse events occurred in all cases and no abnormal change was found in routine blood,liver and kidney function.Conclusion The UC-MSCs infusion is has good safe for treating refractory RA and improves the disease condition to some extent.

Resistance Reversion of Rapamycin on Ovarian Cancer Cell Line SKOV3/DDP and Its Molecular Mechanisms / 中山大学学报(医学科学版)

[Objective]To study the resistance reversion of rapamycin on ovarian cancer cell line SKOV3/DDP,and ex-plore its underlying molecular mechanisms.[Methods]MTT method was used to detect the cell toxicity,drug-resistant multi-ple and reversing multiple of cisplatin-resistant ovarian cancer cell line SKOV3/DDP;Western blot was used to detect the changes of Akt/mTOR Pathway induced by rapamycin.[Results]① MTT detected that when rapamycin concentration was 25,50,100,500 and 1 000 μg/L,its inhibition rates on cisplatin-resistant ovarian cancer cell line SKOV3/DDP were 4.48%,25.30%,35.86%,67.82%,81.43%.The concentration of 25 μg/L was selected to be the reversal concentration,be-cause its maximum rate was less than 5%.②The resistant index(RI)of cisplatin-resistant ovarian cancer cell line SKOV3/DDP was 2.21. ③ The reversal fold of 25 μg/L rapamycin on cisplatin-resistant ovarian cancer cell line SKOV3/DDP was 1.63.④Western blot results:After the addition of rapamycin,expression of p-mTOR and its downstream protein p-p70s6k in SKOV3 and SKOV3/DDP was significantly reduced. Meanwhile,there was a feedback increase in p-Akt.[Conclusions]Rapamycin has a reversal effect on cisplatin-resistant ovarian cancer cell line SKOV3/DDP. Its reversal mechanism may be inhibiting the cell proliferation and promoting cell apoptosis by depressing the expression of p-mTOR and its downstream pro-tein p-p70s6k in Akt/mTOR Pathway.

Comparison of three minimally invasive surgical methods in treatment of impacted upper-ureteral calculi / 中华全科医师杂志

One hundred and fifty patients with impacted upper-ureteral calculi were treated with ureteroscopic lithotripsy (URSL group,n =58),percutaneous nephrolithotomy (PCNL group,n =42) or retroperitoneal laparoscopic ureterolithotomy (RPLU group,n =50) from April 2011 to August 2015.The operative time,length of postoperative hospital stay,stone-free rate and the incidence of postoperative complications were observed and compared among three groups.The stone-free rate of URSL group one month after surgery was 77％,which was significantly lower than PCNL (95％) and RPLU groups (100％) (P ＜ 0.05).But the operative time and length of postoperative hospital stay of URSL group [(37.2 ± 5.6)min,(3.9-±0.8)d] were significantly shorter than PCNL group[(75.2 ±6.7) min,(7.9 ±0.9))d] and RPLU group [(65.3 ± 6.1) min,(8.7 ± 0.9) d] (all P ＜ 0.05).The postoperative complication rates of three groups were 8％,10％ and 10％,respectively.

miR-496 over-expression inhibits growth and metastasis in colon cancer cells / 中国病理生理杂志

[ ABSTRACT] AIM: To investigate the effect of miR-496 over-expression on the growth and metastasis of colon cancer cells and its molecular mechanism .METHODS:The proteins interacting with miR-496 were screened by bioinfor-matic method.The levels of miR-496, CTNNB1 mRNA and β-catenin protein in colon cancer cell lines , HT29, HCT116 and SW480, and normal colonic epithelial cell line NCM 460 were detected by real-time PCR and Western blot .HT29, HCT116 and SW480 cells were transfected with miR-496 mimics using Lipofectamine 2000 and named as HT29-miR-496 mimics, HCT116-miR-496 mimics and SW480-miR-496 mimics cells, respectively, and the cells transfected with the scramble served as negative control .The cell viability, lactate dehydrogenase (LDH) leakage, and colony formation and metastatic abilities were determined by MTT assay , LDH assay, colony formation assay and Transwell method , respective-ly.The promoter activity of miR-496 was measured using luciferase reporter gene assay .The protein levels of β-catenin, eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1), p-4E-BP1, low-density lipoprotein receptor-related protein 6(LRP6), p-LRP6, MMP-7, MMP-9, MMP-13 and TIMP-2 were monitored by Western blot.RESULTS:Endog-enous miR-406 interacted with β-catenin was found in the colon cancer cells .Low miR-496 expression in the HT29, HCT116 and SW480 cells and high miR-496 expression in NCM460 cells were detected.In contrast, high β-catenin ex-pression was found in the HT29, HCT116 and SW480 cells and low β-catenin expression was observed in the NCM460 cells.Compared with control group , the cell viability, colony formation rate and the number of metastatic cells remarkably decreased in the HT29-miR-496 mimics, HCT116-miR-496 mimics and SW480-miR-496 mimic cells (P<0.05).The promoter activity of miR-496 was significantly increased in colon cancer cells transfected with miR-496 mimics, and was 1.75, 2.04 and 1.61 times as high as control group .miR-496 over-expression inhibited β-catenin levels, and p-4E-BP1 and p-LRP6 protein levels were also reduced .siRNA-or over-expressed miR-496-mediated β-catenin down-regulation in-hibited MMP-7 and MMP-9 expression , but promoted TIMP-2 expression .CONCLUSION:The expression level of miR-496 in the colon cancer cells is low , but in the normal colonic epithelial cells is high .miR-496 over-expression inhibits the protein levels of MMP-7 and MMP-9, and promotes the protein expression of TIMP-2 via inhibiting Wnt/β-catenin path-way, thus suppressing malignant phenotype in the colon cancer cells .

Implantation of plate and hollow screw for the repair of the split fracture of greater tuberosity of humerus:comparison of shoulder joint function / 中国组织工程研究

BACKGROUND:Canada Montreal Scholar Mutch et al have recently proposed a new morphologic classification of fracture of greater tuberosity of humerus. They divided these fractures into three typeavulsion, split and depression. OBJECTIVE:To compare the recovery of shoulder function after conventional plate-screw and hol ow-screw fixation for the repair of the split fracture of greater tuberosity of humerus. METHODPatients with greater tuberosity of humerus, who were treated in the Department of Orthopedics of Yichang Yiling Hospital, China from January 2010 to January 2014, were classified according to Mutch’s classification. A total of 83 patients with split greater tuberosity of humerus after complete fol ow-up were retrospectively analyzed. Of them, 23 cases received plate-screw fixation as plate-screw group, and 60 cases received hol ow-screw fixation as hol ow-screw group. Visual Analog Scale, the United States Scores of Shoulder and Elbow Surgeons, and Constant and Murley Scoring Systems were utilized to assess the therapeutic outcomes. Patient’s pain and changes in shoulder function were analyzed before and after treatment. RESULTS AND CONCLUSION:A total of 83 patients were fol owed up. Fixator was obtained at 1 year after surgery in al patients. No significant difference in Visual Analog Scale, the United States Scores of Shoulder and Elbow Surgeons, and Constant and Murley Scoring Systems was detected in both groups before treatment (P>0.05). Significant differences in Visual Analog Scale, the United States Scores of Shoulder and Elbow Surgeons, and Constant and Murley Scoring Systems were detectable in both groups at 16 months after removal of the fixator (P<0.05). Data were better in the hol ow-screw group than in the plate-screw group. Above results suggested that hol ow-screw fixation in the repair of split fracture of greater tuberosite of humerus is simple to be operated, with smal trauma, and is an ideal fixation method. Clinical repair effect is better than plate-screw fixation.

Gelatin sponge impregnated with hemocoagulase reduces postoperative blood loss of patients with lumbar fractures undergoing posterior spinal decompression / 中国组织工程研究

BACKGROUND:A single hemostatic material has been proved not to facilitate wound healing or to produce certain adverse reactions; while composites composed of two or three different materials can improve the advantage and histocompatibility of hemostatic materials. OBJECTIVE:To investigate the effect of gelatin sponge impregnated with hemocoagulase solution on amount of bleeding in patients with lumbar fractures undergoing posterior spinal decompression. METHODS: Fifty patients with lumbar fractures who were scheduled for open reduction, pedicle screw fixation and laminectomy were enroled, including 25 cases treated with gelatin sponge impregnated with hemocoagulase before surgical incision closure as test group and 25 cases treated with single gelatin sponge before surgical incision closure as control group. Postoperative drainage volume, drainage time, length of stay, number of re-admissions and postoperative complications were compared between the two groups. RESULTS AND CONCLUSION:The postoperative drainage volume, drainage time and length of stay in the test group were significantly less than those in the control group (P re-admission of patients was found, and there was no hemocoagulase-impregnated absorbable gelatin sponge- related adverse reaction. These findings indicate that posterior laminectomy with hemocoagulase-impregnated gelatin sponge can significantly reduce patients’ postoperative wound drainage and shorten the length of stay.

Impact of ischemic preconditioning on dynamics of homing of endothelial progenitor cells after renal ischemia reperfusion injury / 中华肾脏病杂志

Objectives To investigate the impact of ischemic preconditioning (IPC) on dynamics of homing of endothelial progenitor cells (EPCs) after renal ischemia reperfusion injury (IR).Methods Sixty male Sprague-Dawley rats were randomly divided into three groups after right-side kidney nephrectomy:for sham-operated rats,lumbotomy without vascular clamping was performed; IR rats were clamped renal blood vessels for 40 minutes while IPC rats were pre-treated with 15 min ischemia and 10 min reperfusion.At 3,12,24 h,and 3 days after reperfusion,the pool of circulating,kidneys,lungs and spleens were harvested.The extent of renal injury was assessed by biochemical and histological examination.The dynamics of homing of EPCs was observed by flow cytometry.Results The rats in IPC group exhibited significant improvements in renal function and morphology.Compared with IR group and sham group,the number of EPCs in blood was increased in the IPC group at 12 h and 24 h after reperfusion (P ＜ 0.05).The number of EPCs in kidney was increased at all times pointin the IPC group and IR group as compared to the sham group (P ＜ 0.05.In addition,EPCs number was increased in IPC group compared with the IR group at 12 h and 24 h [(11.36±0.66)％ vs (6.37±0.69)％,(6.31±0.70)％ vs (4.40±0.60)％,all P＜ 0.05].Compared with IR group and sham group,the number of EPCs in the lung was increased in the IPC groups at 12 h after reperfusion [(2.95±0.66)％ vs (1.78±0.59)％,(1.66±0.61)％,all P ＜ 0.05].The number of EPCs in spleen was increased in the IPC group at 72 h as compared with the IR group and sham group [(0.55±0.06)％ vs (0.34±0.07)％,(0.31±0.06)％,all P ＜ 0.05].Conclusions Endogenous EPCs may home to injured kidney after IPC.EPCs can also gather in the lungs and spleen.

Logistic regression analysis of damp-heat and cold-damp impeding syndrome of rheumatoid arthritis: a perspective in Chinese medicine / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate a method for quantitative differential diagnosis of damp-heat and cold-damp impeding syndrome of rheumatoid arthritis (RA) in Chinese medicine (CM).</p><p><b>METHODS</b>Laboratory parameters were collected from 306 patients with RA. The clinical symptoms and laboratory parameters were compared between patients with these two syndromes (158 with RA of damp-heat impeding syndrome, and 148 with RA of cold-damp impeding syndrome), and a regression equation was established to facilitate discrimination of the two RA syndromes.</p><p><b>RESULTS</b>There were significant differences in disease activity score in 28 joints [DAS28 (4)], erythrocyte sedimentation rate (ESR), white blood cell count (WBC), C-reactive protein (CRP), platelet count (PLT), albumin (ALB) and globulin (GLB) between the two syndrome of RA (P<0.05). Logistic regression analysis showed that the parameters ESR, WBC, CRP, joint pyrexia, joint cold, thirst, sweating, aversion to wind and cold, and cold extremities were statistically useful to discriminate damp-heat from cold-damp impeding syndrome. The regression equation was as follows: P=1/{1+exp[-(3.0-0.021X (1)-0.196X (2)-0.163X (3)-1.559X (4)+1.504X (5)-0.927X (6)-1.039X (7)+1.070X (8)+1.330X (9))]}. The independent variables X (1)-X (9) were ESR, WBC, CRP, hot joint, cold joint, thirst, sweating, aversion to wind and cold, and cold limbs. A P value > 0.5 signified cold-damp impeding syndrome, and a P value < 0.5 signified damp-heat impeding syndrome. The accuracy was 90.2%.</p><p><b>CONCLUSION</b>The regression equation may be useful for discriminating damp-heat from cold-damp impeding syndrome of RA.</p>

Hereditary antithrombin deficiency and hyperhomocysteinemia in venous thromboembolic disease / 中国医学科学院学报

Hypercoagulability state is the result of the interplay of genetic predisposition and risk factors. Many key enzymes and reactions in coagulation and anti-coagulation system are involved. Hereditary antithrombin deficiency is one of the major risk factors of venous thromboembolic disease (VTE), whereas hyperhomocysteinemia may also play a role. This article reviews the recent researches on the contributions of these two factors to VTE.

Construction and identification of p4CCL20-ZsGreen1-DR eukaryotic expression vector / 中国组织工程研究

BACKGROUND: It is necessary to establish a high throughput screening system for anti -inflammatory drugs for rheumatoid arthritis.OBJECTIVE: To construct an eukaryotic expression vector p4CCL20-ZsGreen1-DR with the NF-kB cis-acting element 4×CCL20motif as an enhancer, SV40 as a promoter, and ZsGreen1-DR as a reporter gene.METHODS: The target fragment SV40 was PCR amplified using PGL2-control plasmid as a template. KpnⅠ/Bam HⅠ restriction sites were introduced into the flank of the target fragment. Then, pSV40-ZsGreen1-DR vector was constructed by cloning the target fragment into pZsGreen1-DR plasmid. Finally, p4CCL20-ZsGreen1-DR plasmid was constructed by cloning the double strand DNA of 4×CCL20 motif (with BglⅡ and EcoRⅠ sticky ends at the 5’ and 3’ terminus, respectively) into the corresponding restriction sites of pSV40-ZsGreen1-DR vector (upstream of SV40 promoter).RESULTS AND CONCLUSION: DNA sequencing demonstrated successful construction of p4CCL20-ZsGreen1-DR plasmid.The construction of p4CCL20-ZsGreeR plasmid might be useful to establish a high throughput screening system for anti -inflammatory drugs.

Effects of specific siRNA targeting at bcr-abl fusion gene and its combination with p27 gene clone on chronic myeloid leukemia cell line K562 / 中国实验血液学杂志

This study was purposed to explore the effect of specific small interfering RNA targeting at bcr-abl fusion gene and its combination with p27 gene clone on the proliferation, cell cycle and apoptosis of chronic myeloid leukemia (CML) cell line K562. CML cell line K562 was used as the study object. A 21nt siRNA targeting at the fusion site of b3:a2 mRNA in bcr-abl fusion gene was designed, synthesized and transfected into the K562 cells as RNA interference group. Northern blot was used to detect the bcr-abl fusion gene, Western blot was used to detect the expression of P210 protein and apoptosis-related protein BCL-xL after the transfection. Meanwhile, p27 gene was amplified from peripheral blood mononuclear cells by RT-PCR, and was confirmed to be correct by sequencing, then p27-pcDNA3.1 vector was constructed and transfected into K562 cell line by Lipofectin. After being selected with G418, p27-pcDNA3.1-K562 cell clone stably expressing p27 was isolated. P27 protein was identified by Western blot. Finally, siRNA and p27 gene clone were together applied to K562 cells, the cell survival rate was tested by MTT. The cell cycle and the apoptosis were tested by flow cytometry. The result showed that in contrast with the control group, the expression level of bcr-abl fusion gene was much lower in siRNA group, about 18.4% of K562 cells in siRNA group were apoptotic at 24 hours after siRNA transfection, and the expression of apoptosis-associated protein BCL-xL was greatly down-regulated. The expression of P27 protein could be detected by Western blot in p27-pcDNA3.1-K562 cells. The strong inhibition of cell proliferation was observed in p27-pcDNA3.1-K562 cells, as compared with control K562 cells. The count of p27-pcDNA3.1-K562 cells in G(0)/G(1) phase increased apparently, but that in S phase declined greatly. Cell cycle was arrested in G(0)/G(1) phase. After the combination of p27-pcDNA3.1-K562 cells with specific siRNA, the percentage of apoptosis obviously increased and cell survival rate significantly declined. It is concluded that the specific siRNA distinctly inhibits the expression of bcr-abl fusion gene, and can induce K562 cell apoptosis. The combination of specific siRNA with P27 gene clone displays a synergy of inhibition and pro-apoptosis effects to K562 cells.

Role of mitochondria pathway in signal transduction of chronic myeloid leukemia / 中国实验血液学杂志

This study was aimed to investigate the role of mitochondria pathway in signal transduction of chronic myeloid leukemia (CML). After bcr3/abl2 antisense oligodeoxynucleotide (ASO) was introduced into CML cell line K562 cells by liposomal transfection, the cell viability was detected by MTT assay, the cell apoptosis was determined by flow cytometry (FCM), the mitochondrial membrane potential (DeltaPsi) was labeled by Rhodamine 123 and examined by FCM, and the expression of mitochondrial apoptosis signal transduction pathway related proteins cytochrome C was analyzed by Western blot. The results showed that after K562 cells were exposed to 2 micromol/L of bcr3/abl2 ASO for 24 hours, bcr3/abl2 ASO significantly inhibited cell viability with inhibitory rate of 65.7%, induced the apoptosis of K562 cell line with apoptotic rate of 16.9%, and decreased mitochondrial Deltapsi of K562 cells with the reducing rate of 38.33%, enhanced the expression of cytochrome C with increase of optical density value from 2.33 +/- 0.3 to 4.78 +/- 0.1 by laser photometric scanning. It is concluded that mitochondria pathway plays an important role in signal transduction of chronic myeloid leukemia by directing apoptotic signal transduction.

Effect of tyrosine-kinase Inhibitor on p15 gene transfected K562 cells / 中国实验血液学杂志

The objective of study was to investigate the combined effect of tyrosine-kinase inhibitor (imatinib) and p15 gene on the proliferation, cell cycle and apoptosis of chronic myeloid leukemia cell line K562. p15 gene was amplified from peripheral blood mononuclear cells by RT-PCR, and confirmed by DNA sequencing, then the recombinant p15-pcDNA3.1 vector was constructed and transfected into K562 cell line by Lipofectine. After screening with G418, p15-pcDNA3.1-K562 cell clone stably expressing P15 was isolated. P15 protein was identified by Western blot. The cell survival rate was determined by MTT, cell cycle and apoptosis were detected by flow cytometry. The results showed that partial deletion of p15 gene in K562 cells was verified by DNA sequencing, leading to the function of P15 protein to be lost. The expression of P15 protein can be detected by Western blot in p15-pcDNa3.1-K562 cells. A strong inhibition of cell proliferation was observed in p15-pcDNA3.1-K562 cells as compared with that of the control K562 cell. The cells of G(0)/G(1) phase in p15-pcDNA3.1-K562 cells increased apparently, and S phase cells declined signifcantly. Cell cycle was arrested in G(0)/G(1) phase. The percentage of apoptotic cells greatly increased after transfection with p15-pcDNA3.1-K562 cells combined with imatinib, and cell survival rate notably declined. It is concluded that the imatinib in combination with the expression of p15 gene has a synergistic effect on the inhibition of K562 cell proliferation and promotion of its apoptosis.

Effect and mechanism of sinomenine on the signal transduction of the synovial cell nuclear factor-KB in rats with adjuvant arthritis / 中国组织工程研究

BACKGROUND: Sinomenine is an alkaloid monomer extracted from a Chinese medicinal herb sinomenium acutum stem. It is used in the therapy of the rheumatoid arthritis and has clear and definite therapeutic effects, but the therapeutic mechanism is unclear.OBJECTIVE: To observe the effect of sinomenine at different doses in vitro on the activity of nuclear factor-κB(NF-κβ) and mRNA expressions of the tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) andinterleukin-10 (IL-10) in the synovial cells of the rats with adjuvant arthritis(AA) to explore the probable mechanism of sinomenine in the treatment of rheumatoid arthritis(RA).DESIGN: A controlled repeated measuring study based on the cells.SETTING: Department of traditional chinese medicine and the institute of burn research of a military medical university.MATERIALS: This study was finished at the Laboratory of the Institute of Burn Research of Chinese PLA. The experimental animals were 25 healthy male Wistar rats of clean grade. The AA model rats were made and the synovial cells were collected and grouped as follows: normal control group, AA group,AA + sinomenine 30 mg/L group, AA + sinomenine 60 mg/L group, AA + sinomenine 120 mg/L group. The activity of the NF-κB was measured by the electrophoresis mobility shift assay(EMSA) . The mRNA expressions of the TNF-α, IL-1β and IL-10 were measured by reverse transcription-PCR assay.MAIN OUTCOME MEASURES: The results of the changes of the activity of the NF-κB and the mRNA expressions of the TNF-α, IL-1β and IL-10 in the synovial cells of the rats with adjuvant arthritis after the treatment with sinomenine at different doses.RESULTS: Compared with the normal control group, the activity of the NF-κB and the mRNA expressions of the TNF-α, IL-1β and IL-10 in the synovial cells in the AA group all increased significantly and the outcomes were 17±6, 0.570±0.047, 0.730±0.093, 0. 683 ±0.081 (t= 2.71 -4.07, P < 0.05). After the administration of sinomenine, the activity of NF-κB showed a good correlation with mRNA expressions of the TNF-αandIL-13(r=0.810, P <0.001; r=0.562, P <0.05), but no statistical relevance with mRNA expression of IL-10 was established. Sinomenine showed a dose-dependent inhibition on the activity of the NF-κB and the mRNA expressions of the TNF-α and IL-1β in a certain range of concentrations(30-120 mg/L), but no dose-dependent inhibition on mRNA expression of the IL-10 was observed.CONCLUSION: Through the inhibition of the activity of the NF-κB,sinomenine decreased the mRNA expressions of the TNF-α and the IL-1β in the synovial membrane cells.