Objective To observe the specific expression of diphtheria toxin A chain gene(DTA) in Ewing's sarcoma nude mice models.Methods pS2DTA plasmid was constructed by the Lus gene in pS2 plasmid was replaced with the DTA gene in pIBI30 plasmid,and transformed into DH5?, then amplified.Five nude mice were injected with 2.4?10~(7) SKES1 cells,sacrificed on week 6,and the tumor tissues were implanted onto the back of another 20 nude mice.Till 6 weeks later the diameter of the tumor up to 0.8-1.0 cm,the Ewing's sarcoma nude mice models were successfully established and randomly assigned to receive the injection of 300 ?g lipidosome+100 ?l normal saline(control group) or 300 ?g lipidosome+100 ?l normal saline+100 ?g pS2DTA (experimental group) once a day for 5 d.On day 4 after the therapy finished,4 nude mice of each group were randomly sacrificed and the DTA expression in the tissue of tumor,liver and myocardium was detected by the immunohistochemistry and RTPCR.Results The immunochemical results showed DTA expression in the tissues of control group was all negative while the DTA expression in the experimental group was negative in the tissues of myocardium and liver,positive in tumor tissue.RTPCR showed DTA expression in control group was negative in all tissues while strongly positive in tumor tissue,weakly positive in liver tissue and negative in myocardiac tissue.Conclusion The pS2DTA regulated by the EWSFLI1 fusion gene has a specific expression in transplanted Ewing's sarcoma in mice.

Objective To construct an expression vector containing Ewing's sarcoma EWS FLI 1 specific binding sequence and diphtheria toxin A chain (DTA) gene sequence to investigate the killing effect of DTA on cultured Ewing's sarcoma cells so as to provide proof for the exploration of new methods for gene therapy for cancer. Methods Expression vector (pS2 DTA) containing EWS FLI 1 specific binding sequence and DTA gene sequence was constructed by replacing the luciferase gene in pS2 with DTA gene at the points of Nco Ⅰ and Xba Ⅰ by molecular technique. After transfection of pS2 DTA into Ewing's sarcoma cells and the control cells, DTA expression and its killing effect on cells were detected. Results pS2 DTA was highly expressed in Ewing's sarcoma cell line, and the killing effect of DTA was much higher than that in the control cells. Conclusion pS2 DTA has selective killing effect on Ewing's sarcoma cells and can inhibit the growth of Ewing's sarcoma cells. So pS2-DTA has potential value in the treatment of Ewing's sarcoma.

Objective: To construct a liver targeting gene transfer vector using hepatitis B virus envelope particles. Methods: Hepatitis B viruses were obtained from the supernatant of HepG 2.2.15 cells by a PEG8000 system and were inactivated by ?-propiolactone to prepare hepatitis B virus envelope. The hepatitis B virus envelope was used to pack 5.3 kb pIRES_2-EGFP to assess their packing ability. Subsequently, the products were studied with ELISA, PCR, SDS-PAGE, and electron microscopy. Finally, the product was used to transfect HepG2 cells and the green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer.Results: The acquired hepatitis B virus envelope retained the surface protein HBsAg+pre S_1+pre S_2, but with no virus DNA. The prepared envelope had high packing ability for GFP and the packed GFP had a high transfection rate in HepG2 cell. Conclusion: Hepatitis B virus envelope has been successfully obtained from the supernatant of HepG 2.2.15 cells with a PEG8000 system and ?-propiolactone.

Objective To identify and classify six isolates of swine-originated Trichinella from China. Methods Five specific pairs of primers were synthesized based on DNA sequence of expansion segment V region and internal tran-scribed spacers (ITS1 and ITS2) of ribosomal DNA repeat from Trichinella. International reference strains of five Trich-inella species [Trichinella spiralis (T1), T. nativa (T2), T. britovi (T3), T. pseudospiralis (T4) and T. nelsoni(T7)] were used as control. Six swine Trichienlla isolates from Henan, Yunnan, Harbin, Tongjiang of Heilongjiang, Hubei and Tianjin were identified by multiplex PCR and its effecting factors of PCR amplification were observed. Results Electrophoresis results of multiplex PCR products of Trichinella larvae showed that the band (173 bp) of the six isolates was the same as T. spiralis(T1). The specific band (173 bp) was detected by multiplex PCR through amplification from issues of single T. spiralis larva, the larvae conserved in 80% ethanol for 6 months, the larvae stored in 10% formaldehyde, in 0.05% formaldehyde, 0.2% sodium azide or 0.05% merthiotate for 2 weeks,or fresh mouse muscle with larvae. Conclusion All the six swine Trichinella isolates are identified as T. spiralis (T1) by multiplex PCR.

Objective: To summarize the application of distal fixated long stem in the treatment of intertrochanteric fracture (ITF) in the sequence of proximal femoral reconstruction, fixation materials, and other details after operation, in order to improve doctor's attention to the reconstruction of the proximal femur and reduce complications. Methods: The related literature about the application of distal fixated long stem in the treatment of ITF was extensively reviewed, summarized, and analyzed. Results: The sequence of reconstruction is divided into fracture reconstruction priority and prosthesis reconstruction priority. The former is mainly to provide an anatomical reference for the placement of joint prostheses, the latter is mainly to provide support for fracture fixation. The distal fixated cement long stem and cementless long stem have their own characteristics, and materials of reconstruction are used in combination. There is no uniform standard for the sequence and materials of reconstruction. Conclusion: Although the stability of the distal fixated long stem depends on the distal femur, the accurate reconstruction of the proximal femur is still worthy of attention.

Recently the maintenance therapy of non-small-cell lung cancer (NSCLC) patients who completed required treatment cycles has caused widespread interests in the medical field. Traditional Chinese medicine may be a useful complement in maintenance treatment of mid-to-late stage NSCLC.

Objective:To investigate the clinical effect of long-stem hemiarthroplasty in the treatment of unstable intertrochanteric fracture in elderly patients with severe osteoporosis.Methods:A retrospective analysis was performed on 48 elderly patients with unstable intertrochanteric fractures of the femur with severe osteoporosis in Liaocheng People′s Hospital from April 2017 to April 2019.Twenty three patients received long-stem hemiarthroplasty (LHA group). Twenty five patients were treated with proximal femoral nail anti-rotation (PFNA) (PFNA group). PFNA group was used as the control group.The operative time, intraoperative blood loss, perioperative blood transfusion volume, number of intraoperative fluoroscopy, weight-bearing time after operation, the incidence of postoperative complications, hospitalization time, and Harris hip score of 1, 3, 6, 12 months after surgery, to investigate the efficacy of the application of long-stem hemiarthroplasty.Results:In LHA group, 23 patients were followed up for (18.6±3.9) (range from 12.0 to 26.0) months, and 25 patients in the PFNA group were followed up for (17.8±3.3)(range from 12.0 to 24.0) months.There was no significant difference in follow-up time between the two groups ( Z=-0.552, P=0.581). The operation time of LHA Group (60 (55, 73) h) was longer than that of PFNA Group (55 (50, 60) h). The intraoperative blood loss in LHA Group ((179.35±63.47) mL) was more than that in PFNA Group ((122.80±49.03) mL). The number of fluoroscopy in LHA Group (2 (2, 2) times)was less than that in PFNA Group (16 (14.5, 19.5) times). The time of weight bearing in LHA Group (4 (3, 5) d) was earlier than that in PFNA Group (33 (30, 36) d), and the differences were statistically significant ( Z=2.459, t=3.470, Z=6.216, Z=5.959; all P<0.05). There were no significant differences in perioperative blood transfusion, hospital stay and postoperative complications between the two groups (all P>0.05). Harris hip function score was significantly higher in LHA Group ((76.70±5.96), (82.13±6.38), (85.96±7.16), (88.78±7.67) points) and PFNA Group ((63.80±3.46), (71.56±2.55), (81.60±3.38), (88.08±4.83) points) increased gradually with the increase of follow-up time ( Fintra-group=432.557, Pintra-group<0.001), and the score reached the highest 12 months after operation.Harris hip function score of LHA group was higher than that of PFNA group( Finter-group=25.437, Pinter-group<0.001). There was interaction effect between follow-up time point and operation mode( Finteraction=53.464, Pinteraction<0.001). Conclusion:For the elderly patients with unstable intertrochanteric fracture with severe osteoporosis, the application of lengthened stem hemiarthroplasty can get out of bed early, reduce the complications of bed rest, reduce the number of intraoperative fluoroscopy, and recover the function of hip joint earlier and better with satisfactory results.

Objective: To study the survival time and influencing factors of HIV/AIDS cases who began receiving antiretroviral therapy (ART) from 2005 to 2015 in Tianjin. Methods: Data related to HIV/AIDS cases that receiving ART between 2005 and 2015 in Tianjin, were collected from the Chinese HIV/AIDS Basic Information Management System. A retrospective cohort study was conducted to analyze data of collection. Life table was used to calculate the survival proportion and Cox proportion hazard regression model was used to analyze the factors associated to the time of survival. Results: A total of 2 057 HIV/AIDS cases were involved, including 51 died from AIDS related disease, ending up with the survival rates of 1, 3, 5 and 10 years as 98.4%, 97.8%, 97.4% and 95.8%, respectively. Results from the multivariate Cox proportion hazard regression model showed that when comparing with the cases aged<30 years, aHR (95%CI) of the cases aged 30-39 years, 40-49 years, 50 years or above appeared as 4.506 (1.226-9.059), 5.944 (1.479-13.892) and 15.958 (5.309-27.206) respectively. When comparing with the cases having no loss of follow-up during ART process, the aHR of the cases having lost of follow-up during ART was 5.645 (95%CI: 3.124-10.200). When comparing with the cases diagnosed by other institutions, the aHR of the cases diagnosed by hospitals was 3.823 (95%CI: 1.423-10.274). When compared with the cases had no hepatitis B or hepatitis C before ART, aHR of the cases with hepatitis B or C prior to ART was 2.580 (95%CI:1.210-5.502). Compared with the cases receiving ART at Ⅰ/Ⅱ clinical stages, the aHR of the cases at Ⅲ/Ⅳ clinical stages was 3.947 (95%CI: 2.167-7.188). Compared with the cases with junior high school education or below, the aHR of the cases with high school education or above was 0.440 (95%CI: 0.238-0.810). Compared with the cases diagnosed before operation, aHR of the cases from special investigation and from counseling and testing (VCT) were 0.111 (0.027-0.456) and 0.182 (0.049-0.674) respectively. Conclusions The survival rate of HIV/AIDS cases that received ART was high in Tianjin. Risk factors related to the survival of cases would include: old age when started receiving ART, loss of follow-up during ART, diagnosed by hospitals, co-infected with hepatitis B or hepatitis C and receiving ART at Ⅲ/Ⅳ clinical stages. Meanwhile, protective factors related to the survival of cases would include: having high school or above education, diagnosis was made through other special programs or from VCT services.

Objective To analyze the survival and influencing factors of treatment-naive HIV／AIDS patients aged 15 years or above in Tianjin.Methods The data of 973 untreated HIV／AIDS patients aged 15 years or above who were diagnosed during June 28 1996 to May 28 2017 in Tianjin were retrospectively analyzed.The survival rate of patients was calculated by the life table method , and the factors affecting the survival time were analyzed by the Cox proportional hazard model.Results There were 726 HIV carriers (74.6%) and 247 AIDS patients (25.4%).Among them, there were more males (93.7%, 912／973) than females (6.3%, 61／973); Han nationality accounted for 92.5%(900／973); the education level of most patients was high school or below (71畅0%, 691／973); half of the patients were unmarried (50.4%, 490／973); 57.7%(561／973) of the patients were infected by men who have sex with men ( MSM);62畅9%(612／973) were local residents and 37.1%(361／973) were migrants; 36.6%(356／973) cases were detected by pre-transfusion tests, and 74.5%( 725／973) had initial CD4 +T count tests.In this series, the average survival time is 11.03 years after diagnosis (95%CI 10.01-12畅05 years).The 1, 3, 5, and 10-year survival rates after diagnosis were 96.8%, 88.7%, 80.2%and 40.5%.183 cases (18畅8%) died from AIDS.Cox proportional hazard model analysis showed that compared with patients aged 15 to 35 years, patients aged 35-50 years (HR＝3.077, 95%CI 1.868-5.070) and ＞50 years (HR＝2.626, 95%CI 1畅553-4.440) had higher risk of AIDS-related deaths; compared with high school education or below , patients with college education or above ( HR＝0.562, 95%CI 0.371-0.851) had a lower risk of AIDS death; injection drug user (IDU) (HR＝0.751, 95%CI 0.316-0.909) had lower risk of death than MSM. Patients detected by the consultation ( HR＝0.463, 95%CI 0.252-0.851), special investigation ( HR＝0畅263, 95%CI 0.116-0.593), detainee physical examination ( HR＝0.055, 95%CI 0.007-0畅441) and STD clinic visits (HR＝0.033, 95%CI 0.005-0.239) had a lower risk of death than those detected by pre-transfusion tests.The initial CD4 +T lymphocyte counts were negatively correlated with the risk of AIDS-related death (P＜0.01).Conclusions The average survival time after diagnosis of untreated HIV／AIDS patients aged 15 years or above was 11.03 years in Tianjin.The age, education level, route of infection, detection of HIV infection and the initial CD 4 +T lymphocyte counts are related to the survival time after diagnosis of untreated HIV／AIDS patients.

Objective:To investigate the effect of penehyclidine hydrochloride on respiratory mechanics, arterial blood gas, and inflammatory factors in patients undergoing one-lung ventilation during thoracoscopic lobectomy.Methods:A total of 100 patients who underwent thoracoscopic lobectomy with one-lung ventilation at Jinhua Central Hospital from January to November 2022 were included in this randomized controlled study. They were divided into groups A and B ( n = 50 per group) using a random digital number table. Patients in group A received an intravenous infusion of 0.02 mg/kg of pentylenethyclidine hydrochloride 30 minutes before surgery, while patients in group B received an equal amount of 0.9% sodium chloride injection 30 minutes before surgery. Clinical indicators, respiratory mechanical indicators (peak airway pressure, lung compliance), arterial blood gas analysis indicators (blood oxygen saturation, arterial pressure of oxygen, oxygenation index), inflammatory factor levels (interleukin-6, interleukin-8, tumor necrosis factor level-α), and pulmonary complications were compared between the two groups. Results:There were no significant differences in mechanical ventilation time or total infusion volume between the two groups (both P > 0.05). At the end of surgery (T1) and 1 day after surgery (T2), peak airway pressure in group A was (17.43 ± 2.69) cm H 2O and (16.81 ± 2.28) cm H 2O (1 cm H 2O = 0.098 kPa), respectively, which were significantly lower than (19.23 ± 3.40) cm H 2O and (18.29 ± 2.06) cm H 2O in group B, respectively ( t = 2.94, 3.41, P = 0.002, < 0.001). At T1 and T2, lung compliance in group A was (34.67 ± 2.93) cm H 2O and (36.26 ± 3.11) cm H 2O, respectively, which were significantly higher than (32.23 ± 2.85) cm H 2O and (33.84 ± 2.87) cm H 2O in group B, respectively ( t = 4.22, 4.04, P = 0.000, < 0.001). At T1 and T2, blood oxygen saturation, arterial partial pressure of oxygen, and oxygenation index in group A were significantly higher than those in group B ( t = 8.12, 3.07, 10.47, 3.16, 3.81, 4.15, all P < 0.05). At T1 and T2, interleukin-6, interleukin-8, and tumor necrosis factor-α levels in group A were significantly lower than those in group B ( t = 11.67, 13.55, 9.60, 15.71, 6.13, 11.50, all P < 0.001). The incidence of complications in group A was 4% (2/50), which was significantly lower than 16% (8/50) in group B ( χ2 = 4.00, P < 0.05). Conclusion:Penehyclidine hydrochloride has a good effect on respiratory mechanics, arterial blood gas, and inflammatory factors in patients undergoing thoracoscopic lobectomy with one-lung ventilation and thereby deserves clinical promotion.