Objective: To compare quality control (relative purity and specific activity) and process control [plasminogen (Pg) antigen recovery and potency recovery] indexes of samples before and after adding the Q chromatography step to the full chromatography process of human Pg, thereby determining whether the addition of this step could improve Pg recovery by affinity chromatography. Methods: A Q chromatography step was added before the Pg affinity chromatography in the original Pg chromatography process. The loading solution, flow through solution and eluate of Q chromatography and Pg affinity chromatography were collected. The potency of coagulation factor Ⅱ (FⅡ), Ⅶ (FⅦ), Ⅷ (FⅧ), Ⅸ (FⅨ), and Ⅹ(FⅩ) were detected by the coagulation method, the total protein content was detected by the BCA method, and the Pg potency was detected by the chromogenic substrate method. The content of specific plasma proteins was detected by immunoturbidimetry, the potency recovery of coagulation factors was calculated, and the flow direction of coagulation factors was analyzed. The recovery of different plasma protein antigens were calculated, and the distribution of impurity proteins was analyzed. The relative purity and specific activity of Pg, antigen content, and potency recovery in the target fractions were calculated and compared with the original process indicators, so as to determine the effect of adding Q chromatography on the original process. Furthermore, the reproducibility after process modification was assessed. Results: 100% of FⅡ, FⅩ, and FⅨ, 87.81% of FⅧ, and 40.44% of FⅦ in filtered plasma were removed by Q chromatography. The residual FⅦ (53.26%) and FⅧ (13.30%) in Q flow-through fraction were completely removed by Pg affinity chromatography. In both the original process (without Q-chromatography) and the modified process (with Q-chromatography), non-target plasma proteins mainly existed in the flow-through fraction of Pg affinity chromatography. The antigen recovery of IgM, ceruloplasmin (CER), and fibronectin (FNC) in Q-chromatography flow-through fraction were reduced. In contrast, antigen recovery of other plasma proteins [IgG, IgA, Pg, albumin (AlB), alpha-1-antitrypsin (AAT), and fibrinogen (Fg)] were all >90%, which were consistent with the protein composition and proportion in the original affinity chromatography loading solution. Compared with the recovery rate of Pg antigen in the original process (74.4%), the total recovery of Pg antigen in the modified process was significantly increased (89.97%). Compared with the recovery of IgG (97.48%) and Fg (95.32%) in the Pg affinity flows-through fraction of the original process, the modified process resulted in a slight reduction in the recovery of IgG (94.60%), while the recovery of Fg was not affected (95.05%). The potency recovery rate, specific activity, and relative purity of Pg after Q chromatography were 99.3%, 0.016 U/mg, and 0.15%. These values were the same as those of Pg affinity chromatography loading solution by the original process, indicating that introduction of Q chromatography did not affect subsequent Pg affinity chromatography. Compared with the recovery of Pg antigen in three batches of the original process (66.49±1.02)%, the recovery of Pg antigen in the affinity chromatography eluent of the modified process [five batches; (77.43±4.43)%] was significantly improved. Furthermore, the potency recovery was (86.80±4.28)%, the relative purity was (81.99±1.25)%, the specific activity was (8.679±1.073)U/mg, and the process was reproducible. Conclusion: The addition of Q chromatography could improve the recovery of Pg affinity chromatography in the full chromatography process.

Objective:To explore medical students'cognition,current status,and demands regarding practical skills training in a medical college under the background of new medical science.Methods:A cross-sectional survey was conducted among medical undergraduates from Year 1 to Year 5 in a medical college.A self-designed scale was used to investigate the cognition,current status,and demands related to new medical education concepts.Pearson correlation analysis was used to analyze the relationship between satisfaction with practical teaching facilities and variables including understanding of new medical education,curriculum design,and proportion of practical teaching.Multiple linear regression was conducted to investigate the factors influencing satisfaction.Results:Among 1 253 participants,70.71%acquired new medical science information through online media,while 52.99%learned about it via school courses.Over 80%endorsed"whole-cycle health management"(preventive care before disease onset 91.94%,disease treatment 81.72%,and post-illness rehabilitation 82.60%).Experimental courses(85.16%),clinical skills training(66.88%),and social practice(66.24%)were primary practical forms,but the participation rates of research practice(40.78%)and innovation/entrepreneurship practice(34.72%)was comparatively lower.The satisfaction with practical teaching was positively correlated with adequacy of faculty guidance(r=0.707)and curriculum rationality(r=0.522)(P＜0.01).The multiple linear regression analysis showed that faculty guidance(β=0.436)and the proportion of practical teaching(β=0.319)were the key predictors of satisfaction.Conclusion:Medical students'cognition of practical skills training,curriculum optimization,and adequacy of faculty guidance significantly influence satisfaction with practical teaching,with faculty guidance and rationality of practical process playing key roles.

Objective:To explore medical students'cognition,current status,and demands regarding practical skills training in a medical college under the background of new medical science.Methods:A cross-sectional survey was conducted among medical undergraduates from Year 1 to Year 5 in a medical college.A self-designed scale was used to investigate the cognition,current status,and demands related to new medical education concepts.Pearson correlation analysis was used to analyze the relationship between satisfaction with practical teaching facilities and variables including understanding of new medical education,curriculum design,and proportion of practical teaching.Multiple linear regression was conducted to investigate the factors influencing satisfaction.Results:Among 1 253 participants,70.71%acquired new medical science information through online media,while 52.99%learned about it via school courses.Over 80%endorsed"whole-cycle health management"(preventive care before disease onset 91.94%,disease treatment 81.72%,and post-illness rehabilitation 82.60%).Experimental courses(85.16%),clinical skills training(66.88%),and social practice(66.24%)were primary practical forms,but the participation rates of research practice(40.78%)and innovation/entrepreneurship practice(34.72%)was comparatively lower.The satisfaction with practical teaching was positively correlated with adequacy of faculty guidance(r=0.707)and curriculum rationality(r=0.522)(P＜0.01).The multiple linear regression analysis showed that faculty guidance(β=0.436)and the proportion of practical teaching(β=0.319)were the key predictors of satisfaction.Conclusion:Medical students'cognition of practical skills training,curriculum optimization,and adequacy of faculty guidance significantly influence satisfaction with practical teaching,with faculty guidance and rationality of practical process playing key roles.

The study was aimed to assess the impact of Brucea javanica oil emulsion(BJOE)on the responsiveness of programmed death receptor-1(PD-1)monoclonal antibody to lung adenocarcinoma in mice.The experimental approach involved subcutaneously inoculating Lewis's lung adenocarcinoma(LLC)cells into C57BL/6 mice,which were then divided into four groups:model group,25 ml·kg-1 BJOE group,10 mg·kg-1 PD-1 group,and combination group(25 ml·kg-1 BJOE and 10 mg·kg-1 PD-1).Tumor volume,mass,and inhibition rate were evaluated;the apoptosis within tumor tissue was detected by TUNEL staining;CD4+and CD8+T cell proportions within tumor tissues were analyzed by flow cytometry;the levels of tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ)and granzyme B in tumor tissue were measured by enzyme-linked immunosorbent assay(ELISA).Additionally,LLC cells were categorized into a control group and three BJOE treatment groups(10,30,50 μl·ml-1),and programmed death ligand 1(PD-L1)expression in tumor tissues and LLC cells were assessed by Western blotting.Data showed that as compared with the model group,PD-1 monoclonal antibody alone did not significantly alter tumor volume,tumor mass,CD4+and CD8+T cell proportions,cytokine levels(IFN-γ,TNF-α,Granzyme B),or apoptosis in lung cancer-bearing mice.However,BJOE treatment reduced tumor volume and mass,enhanced CD4+and CD8+T cell proportions,increased cytokine levels,and augmented apoptosis(all P<0.05).Furthermore,the combination therapy of BJOE and PD-1 monoclonal antibody yielded significantly greater reductions in tumor volume and mass,with heightened CD4+and CD8+T cell proportions,cytokine levels,and apoptosis compared to either treatment alone(all P<0.05).Both BJOE treatment and the combination therapy significantly upregulated PD-L1 protein expression in tumor tissues compared to the model or PD-1 monoclonal antibody groups(P<0.05).Similarly,BJOE treatment at all tested concentrations significantly increased PD-L1 protein expression in LLC cells as compared to the control group(P<0.05).In conclusion,BJOE could upregulate PD-L1 expression in LLC cells and enhance the sensitivity of lung adenocarcinoma-bearing mice to PD-1 monoclonal antibodies.

Objective:To compare and analyze the clinical efficacy of laparoscopic microwave ablation and laparoscopic hepatectomy in the treatment of hepatic hemangioma.Methods:The clinical data of 98 patients with hepatic hemangioma admitted to the Hepatobiliary Center, the First Affiliated Hospital of Nanjing Medical University from June 2019 to June 2023 were retrospectively analyzed, including 20 males and 78 females, aged 24-69 years. According to the surgical method, they were divided into two groups: laparoscopic microwave ablation group (ablation group) with 34 cases, and laparoscopic hepatectomy group (resection group) with 64 cases. The differences in intraoperative and postoperative recovery related indicators, follow-up and prognosis between the two groups were compared and analyzed.Results:The operative time and blood loss in the ablation group were (90.6±21.8) min and (60.3±40.8) ml, respectively, which were lower than those in the resection group (128.7±30.0) min and (165.8±212.7) ml, and the differences were statistically significant ( t=-6.54, -2.86, P<0.001, P=0.005). There were 5 cases (14.71%) of residual lesions in the ablation group and none in the resection group, with a significant difference between the two groups ( χ2=0.01, P=0.003). The ablation group was superior to the resection group in hospital stay, drainage tube removal time and postoperative pain, with statistical significance (all P<0.05). On the 1st and 3rd day after surgery, the levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin and direct bilirubin in ablation group were lower than those in resection group, and the differences were statistically significant (all P<0.05). In the ablation group, there were 14 cases of hemoglobinuria (41.2%), 2 cases of abdominal hemorrhage (5.9%), 0 cases of bile leakage and 6 cases of pleural effusion (17.7%), while in the resection group, these complications were 2 cases (3.1%), 18 cases (28.1%), 11 cases (17.2%) and 32 cases (50.0%), respectively, and there were statistical significance between the two groups (all P<0.05). In terms of prognosis, there was both one recurrence in each group (2.9% vs. 1.6%), with no significant difference ( χ2=1.00, P=0.653). Conclusion:Compared with laparoscopic hepatectomy, laparoscopic microwave ablation has obvious advantages in terms of operation time, intraoperative blood loss, hospital stay, postoperative pain and complications.

【Objective】 To explore the effects of intraoperative autologous blood(ABT) transfusion on thrombelastography(TEG) in patients underwent neurosurgical procedures. 【Methods】 96 patients (49 males and 47 females) aged 15~79 years who received neurosurgical procedures in our hospital from November 2018 to November 2020 were retrospectively analyzed and divided into autologous blood transfusion group(Group A, n=52)and allogeneic blood transfusion group(Group B, n=44)according to different blood transfusion strategy in operation. The red blood transfusion status, hemoglobin (Hb), hematocrit (Hct), platelet (Plt), fibrinogen(Fib), prothrombin time (PT), activated partial thromboplastin time(APTT), and TEG parameters [activated clotting time(ACT), coagulation time (K), angle rate of clot formation(Angle), maximum amplitude(MA)] before and 1 day after surgery were compared between the two groups. 【Results】 The amount of average blood transfusion didn′t differ significantly by groups (P>0.05). The incidence of extra allogeneic blood transfusion was 17.3%(9/52) in group A, and the amount of average allogeneic blood transfusion in group A was significantly lower than that in group B(333.3±81.7 vs 639.8±258.2, P<0.05). Before operation, the differences in Hb, Hct, Plt, Fib, PT, APTT, ACT, K, MA and Angle levels between the 2 groups were not statistically significant (P>0.05). One day after operation, the Hb(g/L) (109.4±15.8 vs 97.0±15.1), Hct (%) (32.0±4.3 vs 28.3±6.1), Plt(×10⁹/L)(154.2±54.2 vs 120.7±41.6), Fib(g/L)(2.2±0.5 vs 1.6±0.6), MA(mm)(65.0±7.2 vs 60.7±8.7) and Angle levels(deg)(69.1±5.2 vs 62.6±9.8) in group A were significantly higher than those in group B(P<0.05), and the PT(s)(11.9±1.5 vs 12.8±0.9), APTT(s)(27.4±3.3 vs 30.4±5.4), ACT(s)(111.0±14.9 vs 119.1±12.3) and K levels(min)(87.2±25.7 vs 106.4±28.0) in group A were significantly lower than those in group B (P<0.05). 【Conclusion】 Intraoperative ABT in patients underwent neurosurgical procedures can reduce allogeneic blood transfusion, has less effect on coagulation function and TEG, and is safe and effective.

Gene therapy is a rapidly developing field. The most widely used technique for foreign gene transfer is lentiviral-mediated gene therapy. Lentiviral vector has been developed from the first generation to the third generation in terms of safety. The preparation of lentiviruses with high titer remains difficult. In this study, a Fibra-Cel sheet carrier was used as an HEK293T cell carrier matrix, and several sterile cell culture spinners were combined and cultured on a roller bottle machine to scale up the adherent cells. The virus titer was maximized by screening the factors to optimize the lentivirus titer in the third-generation lentivirus packaging process one by one. Fibra-Cel sheet vector was successfully used as the matrix of HEK293T cell adhesion to culture adherent cells at large scale. The optimal conditions for large-scale preparation of the third-generation lentivirus by bottle roller were screened and three batches of lentiviruses were produced on pilot scale. The production time of lentivirus was shortened from 120 hours to 54 hours from plasmid transfection to virus collection; in terms of cost, a rolling bottle machine was used instead of a bioreactor, leading to lower cost and no need for repeated sterilization during the whole process. The safe, effective and low-cost operation of successful production will provide a technical base for the large-scale preparation of lentivirus and thus lay a firm foundation for its clinical application.
Genetic Vectors ; HEK293 Cells ; Humans ; Lentivirus ; Transduction, Genetic ; Transfection

Histone acetylation is one of the most important reactions of post-translational modification of histones, which plays an important role in the regulation of epigenetic processes. Histone deacetylase 2 as a member of type I histone deacetylases,involved in the catalytic regulation of histone and a variety of non-histone deacetylation,regulates a variety of life processes. This paper summarizes the basic structure of histone deacetylase 2 and the role of histone deacetylase 2 in various diseases,and provides a theoretical basis for conducting related studies.

Objective:To observe the sensitivity of transcription mediated amplification (TMA),and to compare its performance with real-time reverse transcription polymerase chain reaction (real-time RT-PCR) in detecting human immunodeficiency virus RNA (HIV RNA).Methods:TMA system was established with TaqMan probes,specific primers,moloney murine leukemia virus (MMLV) reverse transcriptase,T7 RNA polymerase,and reaction substrates.The sensitivity of TMA was evaluated by amplifying a group of 10-fold diluted HIV RNA standards which were transcribed in vitro.A total of 60 plasma of HIV infected patients were measured by TMA and Cobas Amplicor HIV-1 Monitor test to observe the positive rate.The correlation and concordance of the above two technologies were investigated by linear regression and BlandAltman analysis.Results:TMA system was established successfully and HIV RNA transcribed standards at concentration of equal or more than 10 copies/mL could be detected by TMA technology.Among 60 samples of plasma from HIV infected patients,46 were positively detected and 12 were negatively amplified by both TMA and Cobas reagents;2 samples were positively tested by Cobas reagent but negatively tested by TMA system.The concordance rate of the two methods was 97.1％ and the difference of positive detection rate between the two methods was not statistically significant (P＞0.05).Linear regression was used for 46 samples which were positively detected by both TMA and Cobas reagents and showed an excellent correlation between the two reagents (r=0.997,P＜0.001).Bland-Altma analysis revealed that the mean different value ofHIV RNA levels for denary logarithm was 0.02.Forty-four samples were included in 95％ of credibility interval of concordance.Conclusion:TMA system has the potential of high sensitivity.TMA and real-time RT-PCR keep an excellent correlation and consistency in detecting HIV RNA.