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AIM: To study the effects of protein kinase C on the expression of MMPs that may play an important role in the formation of atherosclerosis and the possible mechanism of Danshen injection to treat atherosclerosis. METHODS: 50 Sprgue-Dawley (SD) rats were divided into three groups randomly: control group (group C), model group (group M) and Denshen treatment group (group D). The serum was collected to measure the level of cholesterol, triglyceride, low density lipoprotein cholesterol(LDL-ch). The expression of PKC and MMPs were measured by immunohistochemistry. Light microscope and electron microscope were also used. RESULTS: ① The cholesterol, triglyceride, LDL-ch concentrations in group M and group D were significantly higher than those in group C (P

Objective To evaluate the application effect of pneumatic arm assisted single surgeon uniportal thoracoscopic surgery.Methods A total of 40 patients were admitted to our treatment group in Shanghai Pulmonary hospital in October who accepted pneumatic arm assisted single surgeon uniportal thoracoscopic surgery.All the clinic statistics of patients were collected including operative time,the volume of blood intra-operation,postoperative complications and hospitalization time.23 of 40 patients who accepted lobectomy/segmentectomy with pneumatic arm assisted single surgeon uniportal thoracoscopic surgery were assigned to the observation group,while another 30 concurrent patients who accepted lobectomy/segrnentectomy with conventional uniportal thoracoscopic surgery were assigned to the control group.Both groups were compared.Results The average postoperative hospitalization time of observation group was(4.6 ± 1.3) days.The average time for postoperative drainage tube retention was(46.7 ± 18.6) hours.The average operation time of patients in observation and control groups was(121.74 ± 25.16) min and (119.7 ± 14.26) min separately.The volume of blood intra-operation in observation group was(91.74 ± 32.88)ml and(89.00 ± 41.22) ml in control group.There is no significant difference between two groups(P ＞ 0.05).Conclusion The field of view and adjustment of camera in uniportal thoracoscopic surgery by single surgeon with pneumatic arm assistance are more accurate and steady,in which human resource can be saved.It is safe and reliable and does not prolong operative time or increase bleeding during operation,and can be applied to different kinds of diseases in thoracic surgery.It is worth promotion and application in eligible hospitals and medical institutions.

Objective:To explore the efficacy and safety of transurethral anatomical vapor incision technique of prostate (VIT) with six-step method high power side-emitting greenlight laser in the treatment of benign prostatic hyperplasia (BPH).Methods:A retrospective analysis of 82 patients with BPH who used high power side-out green laser in the treatment from October 2018 to June 2020 in Gongli Hospital of Naval Medical University was performed. Among them, 40 patients were treated with six-step method VIT, and 42 patients were treated with photoselective vaporization of prostate (PVP). The two groups of patients were compared in age [(71.1±8.7)years vs.(72.1±7.0)years], prostate volume [75 (68.25, 89.00) ml vs. 73 (63.25, 85.00) ml], and peak urinary flow rate (Q max) [6.20 (5.20, 8.20) ) ml/s vs. 5.9 (4.75, 7.50) ml/s], post-void residual volume (PVR) [74.00 (42.50, 103.75) ml vs. 67.00 (58.00, 84.50) ml], international prostate symptom score (IPSS) [(21.2±5.2) vs. ( 21.0±3.9)], quality of life score (QOL) [5 (4, 6) vs. 5 (4, 6) ], prostate specific antigen (PSA) [6.20 (4.12, 8.43) ng/ml vs. 5.40 (3.88, 7.13) ng/ml ]. In general, there was no statistical difference ( P>0.05). The VIT group adopts the six-step method of marking, removing film, grooving, excision, trimming and crushing. In the PVP group, the prostate tissue was uniformly vaporized layer by layer from the inside to the outside. Perioperative indexes and complications were compared between the two groups. The Q max, IPSS, QOL, PVR and PSA between the two groups before and 3 months after surgery were compared. Results:All patients in the VIT group and PVP group successfully completed the surgery, and there was no case of transfer to TURP or open surgery. The average operation time was [60.00(50.00, 73.75)min vs. 70.00(50.00, 73.75)min] ( P<0.05). There was no significant difference in the amount of postoperative hemoglobin decline[15.00(10.00, 17.75)g/L vs. 16.00(14.00, 19.25)g/L], average bladder irrigation time[1(1, 1)d vs. 1(1, 1)d], indwelling catheterization time[3(3, 3)]d vs. 3(3, 3)d] and hospitalization time in patients after operation[4(3, 4)d vs. 4(4, 4)d] ( P>0.05). All patients had no blood transfusion, second bleeding, readmission, TURS, urethral stricture and urinary incontinence.There were 2 cases (5.0%) of postoperative urinary tract infection in the VIT group and 9 cases (21.4%) of postoperative urinary tract infection in the PVP group ( P<0.05), and they were cured after anti-inflammatory treatment. Three months after operation, Q max, IPSS, QOL, PVR and PSA in the two groups were significantly improved compared with preoperatively. Among them, the differences of IPSS [(5.7±2.5) points vs. (7.5±2.8) points] and PSA [2.65(2.10, 3.90)ng/ml vs. 4.00(2.45, 4.45)ng/ml] in the VIT group and PVP group after operation were statistically significant ( P<0.05). Conclusions:Applying the six-step method high power side-emitting greenlight laser transurethral anatomical VIT to treat BPH, there is less intraoperative and postoperative bleeding, short operation time, significant decrease in PSA, and fewer complications. It is a safe and effective minimally invasive technology for the treatment of BPH.

A randomized crossover study was performed to compare the effects of low glycemic index diets (LGI)and high glycemic index diets(HGI)on blood glucose,lipid profile and control of body weight in patients with type 2 diabetes.Compared with HGI group,the fasting serum insulin,Homa-IR,LDL-C and body weight significantly decreased in LGI group(P

Objective To establish a pseudovirus system for phenotypic drug-resistance detection and provide a relatively cheap and easy method for drug-resistance testing. Methods EGFP gene was amplified from plasmid pSV-EGFP and then cloned to backbone plasmid pNL4-3.Luc. E-R-by double enzyme digestion;env gene was amplified from RNA isolated from HIV-1-infected persons and cloned to eukaryotic expression plasmid cells and EGFP or ENV expression. Pseudovirus was produced by co-transfection of two recombinant plasmids to 293t cells. Infection of pseudovirus was determined by co-cultured with TZM-b1 cells and immunofluorescent test. Results Two recombinant plasmids(mass ratio,pcDNA3.1-env:pNL4-3.EGFP.E-R-.=2:1)were co-transfected to 293t cells. Cultured supernatants containing pseudovirus were harvested at 48 h post-transfection. Fluorescence was observed in TZM-b1 cells after TZM-b1 cells were infected with pseudovirus at 48 h post-infection. Conclusion The recombinant pseudovirus carrying EGFP gene is constructed successfully and it could be used for phenotypic drug-resistance detection.

OBJECTIVE: To construct an effective lentiviral vector for RNA interference (RNAi) with human glucose transporter 3 (GLUT3)gene.
 METHODS: Four pairs of shRNA sequences against different parts of GLUT3-mRNA were separately cloned into the RNAi plasmid vector pLV-shRNA by recombinant DNA technology to construct shRNA expression vectors pLV-shRNA-GLUT3-1, pLV-shRNA-GLUT3-2, pLV-shRNA-GLUT3-3, and pLV-shRNA-GLUT3-4. The vectors were transfected into HeLa cells to detect the effectiveness of GLUT3 gene silencing. One of effective vectors was selected and co-transfected into 293T cells with lentivirus packaging plasmids to obtain packaged lentivirus particles LV-GLUT3. After viral titer determination, U251 glioblastoma cells were infected with LV-GLUT3 at a multiplicity of infection (MOI) of 10. Finally, the expression of GLUT3 protein was detected by Western blot. 
 RESULTS: DNA sequencing demonstrated that the shRNA sequences were successfully inserted into the pLV-shRNA vectors. In HeLa cells, the expression of GLUT3-mRNA was significantly down-regulated by the recombinant vectors compared with negative control. The recombinant lentivirus LV-GLUT3 harvested from 293T cells had a titer of 1.5×10(9) TU/mL. After infection with LV-GLUT3, the expression of GLUT3 protein in U251 glioblastoma cells was down-regulated. 
 CONCLUSION An effective lentiviral shRNA expression vector targeting the GLUT3 gene is successfully constructed and can be used for further study on the functions of GLUT3 gene.
Genetic Vectors ; Glucose Transporter Type 3 ; genetics ; HEK293 Cells ; HeLa Cells ; Humans ; Lentivirus ; Plasmids ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transfection