Objective:To explore the effectiveness of applying the learning group teaching method based on primary care guidelines for diagnosis and treatment of common diseases in the standardized training of general practice residents (referred to as "general practice resident training").Methods:This study was a randomized controlled trial. A total of 48 trainees enrolled in the general practice residency program at the Affiliated Huai′an No.1 People′s Hospital of Nanjing Medical University from September 2023 to August 2024 were included. They were divided into the observation group and the control group using a random number table method, with 24 trainees in each group. At baseline, general information such as age, sex, and education level of the trainees was collected, and their relevant theoretical knowledge was assessed. The control group trainees were intervened using the traditional teaching method, while the observation group trainees were intervened using the learning group teaching method based on the diagnosis and treatment guidelines for common diseases at primary care level. The teaching activities lasted for 1 year, with sessions held once a month, for a total of 12 sessions. At the end of the intervention, all trainees were assessed on theoretical knowledge, case analysis, and clinical skills. In addition, questionnaires were used separately to survey the trainees′ satisfaction in both groups, as well as the teaching satisfaction of instructors in the observation group.Results:The observation group included 18 females (75%), with a median age of 25 (24, 26) years, while the control group had 16 females (67%), with a median age of 25 (24, 26) years. There was no statistically significant difference in the age, proportion of female trainees, proportion of rural order oriented free medical students, proportion of undergraduate students, proportion of first-year trainees, and theoretical knowledge test scores before the implementation of intervention measures between the two groups of trainees (all P>0.05). At the end of the intervention, the theoretical knowledge, case analysis, and clinical skill test scores of the observation group were higher than those of the control group (all P<0.05). All 48 trainees participated in the survey. The observation group trainees were found to be more satisfied than the control group with the teaching methods, knowledge mastery, clinical diagnosis and treatment thinking, stimulation of learning interest, self-learning ability, teaching interest, interpersonal communication ability, teamwork ability, and community problem-solving ability (all P<0.05). Of the four teachers in the observation group who participated in the satisfaction survey, all expressed satisfaction with the teaching method and its content. They all believed that the teaching method helped them to broaden their teaching ideas and improve their teaching abilities and skills. Conclusions:The learning group teaching method, which is based on the diagnosis and treatment guidelines for common diseases at primary care level, can effectively enhance the theoretical knowledge, case analysis ability, and clinical skills of the trainees. This teaching method is highly effective, with both trainees and teachers expressing great satisfaction.

Objective:To explore the effectiveness of applying the learning group teaching method based on primary care guidelines for diagnosis and treatment of common diseases in the standardized training of general practice residents (referred to as "general practice resident training").Methods:This study was a randomized controlled trial. A total of 48 trainees enrolled in the general practice residency program at the Affiliated Huai′an No.1 People′s Hospital of Nanjing Medical University from September 2023 to August 2024 were included. They were divided into the observation group and the control group using a random number table method, with 24 trainees in each group. At baseline, general information such as age, sex, and education level of the trainees was collected, and their relevant theoretical knowledge was assessed. The control group trainees were intervened using the traditional teaching method, while the observation group trainees were intervened using the learning group teaching method based on the diagnosis and treatment guidelines for common diseases at primary care level. The teaching activities lasted for 1 year, with sessions held once a month, for a total of 12 sessions. At the end of the intervention, all trainees were assessed on theoretical knowledge, case analysis, and clinical skills. In addition, questionnaires were used separately to survey the trainees′ satisfaction in both groups, as well as the teaching satisfaction of instructors in the observation group.Results:The observation group included 18 females (75%), with a median age of 25 (24, 26) years, while the control group had 16 females (67%), with a median age of 25 (24, 26) years. There was no statistically significant difference in the age, proportion of female trainees, proportion of rural order oriented free medical students, proportion of undergraduate students, proportion of first-year trainees, and theoretical knowledge test scores before the implementation of intervention measures between the two groups of trainees (all P>0.05). At the end of the intervention, the theoretical knowledge, case analysis, and clinical skill test scores of the observation group were higher than those of the control group (all P<0.05). All 48 trainees participated in the survey. The observation group trainees were found to be more satisfied than the control group with the teaching methods, knowledge mastery, clinical diagnosis and treatment thinking, stimulation of learning interest, self-learning ability, teaching interest, interpersonal communication ability, teamwork ability, and community problem-solving ability (all P<0.05). Of the four teachers in the observation group who participated in the satisfaction survey, all expressed satisfaction with the teaching method and its content. They all believed that the teaching method helped them to broaden their teaching ideas and improve their teaching abilities and skills. Conclusions:The learning group teaching method, which is based on the diagnosis and treatment guidelines for common diseases at primary care level, can effectively enhance the theoretical knowledge, case analysis ability, and clinical skills of the trainees. This teaching method is highly effective, with both trainees and teachers expressing great satisfaction.

Forty trainees who underwent position-transition training for general practitioner in the Affiliated Huai′an First People′s Hospital of Nanjing Medical University from August 2021 to July 2022 were randomly divided into the control group and the trial group with 20 in each group. The traditional teaching methods were applied in control group and the problem-based learning/case-based learning (PBL/CBL) combined scenario simulation teaching method were applied in trial group. The effect of the two teaching methods was evaluated by the examination results and the teaching satisfaction was assessed by the questionnaire survey. The exam scores of general practice thinking, comprehensiveness of inquiry content, integrity of preliminary diagnosis, comprehensiveness of preliminary treatment, ability to deal with critical and severe cases, ability of chronic disease management and concept of holistic care in trial group were significantly higher than those in control group( t=-15.79,-15.74,-16.67,-13.66,-18.57,-18.55,-19.74,all P<0.05). The satisfaction of trainees in the trial group was also significantly higher than that in the control group in terms of teaching methods, stimulating learning interest; and improvement of independent learning ability, general practice clinical thinking, doctor-patient communication ability, teamwork ability and community problem-solving ability( χ2=12.10,10.03,12.22,10.03,8.18,10.03,8.18,all P<0.05). The study shows that PBL/CBL combined scenario simulation teaching method can improve the learning effect and satisfaction of the trainees in position-transition training for general practitioner.

To modify the splicing pattern of Bcl-x and compare the effect of this approach with that of the antisense gene therapy in BIU-87 cell line of bladder cancer, by using 5'-Bcl-x AS to target downstream alternative 5'-Bcl-x splice site to shift splicing from Bcl-xL to Bcl-xS and 3'-Bcl-x AS antisense to the 3'-splice site of exon III in Bcl-x pre-mRNA to down regulation of Bcl-xL expression, the inhibitory effects on cancer cells by modification of alternative splicing and antisense gene therapy were observed and compared by microscopy, MTT Assay, RT-PCR, FACS, Westhern bloting and clone formation. The growth of cells BIU-87 was inhibited in a dose- and time-dependent manner. Its inhibitory effect began 12 h after the exposure, reaching a maximum value after 72h. The number of cells decreased in S phase and the number increased in G1 phase. The ability to form foci was reduced and the antisense gene therapy was approximately half as efficient as modification of alternative splicing in inducing apoptosis. It is concluded that modification of splicing pattern of Bcl-x pre-mRNA in bladder cancer cell BIU-87 is better than antisense gene therapy in terms of tumor inhibition.

The killing effects of herpes simplex virus thymidine kinase gene/ganciclovir (HSV-tk/GCV) approach by the addition of several commonly clinical chemotherapeutic agents on hormone refractory prostate cancer (HRPC) cells PC-3m were investigated. After transferring of the HSV-tk gene into PC-3m cells, mRNA and protein expression of HSV-tk was detected by reverse-transcript polymerase chain reaction (RT-PCR) and strept avidin-biotin complex (SABC) immunohistochemical method. The killing effect of GCV, cisplatin (CDDP), etoposide (VP-16), vincristine (VCR), methotrexate (MTX), 5-fluorouracil (5-Fu), and suramin on PC-3m cells was evaluated by morphological assessment analysis, trypan blue exclusion assay and MTT assay respectively. Additionally, the cooperative effect of HSV-tk/GCV system combined with the above agents on the target cancer cells was determined by MTT. Furthermore, apoptosis and necrosis induced by GCV plus 5-Fu or suramin was analyzed by flow cytometry (FCM). The results showed that that there was HSV-tk mRNA and protein expression in pDR2-tk plasmid transduced PC-3m cell. Combination of GCV with VP-16, VCR, 5-Fu or suramin led to an enhanced cellular killing effect, but with CDDP resulted in a reduced one and with MTX in an approximate one. FCM revealed that synergistic use of GCV and 5-fu or suramin resulted in a rather large proportion of apoptosis and necrosis with the apoptosis index being 36.38% and 35.51%, and the proportion of necrosis being 33.05% and 28.87%, respectively. In conclusion, HSV-tk/CGV approach by addition of certain clinical available chemotherapeutic drugs brings on statistically significant enhanced cell killing over single-agent treatment. Our results highlight the potential for such new combination therapies for future treatments of HRPC.

To modify the splicing pattern of Bcl-x and compare the effect of this approach with that of the antisense gene therapy in BIU-87 cell line of bladder cancer, by using 5'-Bcl-x AS to target downstream alternative 5'-Bcl-x splice site to shift splicing from Bcl-xL to Bcl-xS and 3'-Bcl-x AS antisense to the 3'-splice site of exon Ⅲ in Bcl-x pre- mRNA to down regulation of Bcl-xL expression,the inhibitory effects on cancer cells by modification of alternative splicing and antisense gene therapy were observed and compared by microscopy, MTT Assay, RT-PCR, FACS, Westhern bloting and clone formation. The growth of cells BIU-87 was inhibited in a dose- and time-dependent manner. Its inhibitory effect began 12 h after the exposure, reaching a maximum value after 72h. The number of cells decreased in S phase and the number increased in G1 phase. The ability to form foci was reduced and the antisense gene therapy was approximately half as efficient as modification of alternative splicing in inducing apoptosis. It is concluded that modification of splicing pattern of Bcl-x pre-mRNA in bladder cancer cell BIU-87 is better than antisense gene therapy in terms of tumor inhibition.

The killing effects of herpes simplex virus thymidine kinase gene/ganciclovir (HSV-tk/GCV) approach by the addition of several commonly clinical chemotherapeutic agents on hormone refractory prostate cancer (HRPC) cells PC-3m were investigated. After transferring of the HSV-tk gene into PC-3m cells, mRNA and protein expression of HSV-tk was detected by reverse-transcript polymerase chain reaction (RT-PCR) and strept avidin-biotin complex (SABC) immunohistochemical method. The killing effect of GCV, cisplatin (CDDP), etoposide (VP-16), vincristine (VCR), methotrexate (MTX), 5-fluorouracil (5-Fu), and suramin on PC-3m cells was evaluated by morphological assessment analysis, trypan blue exclusion assay and MTT assay respectively. Additionally, the cooperative effect of HSV-tk/GCV system combined with the above agents on the target cancer cells was determined by MTT. Furthermore, apoptosis and necrosis induced by GCV plus 5-Fu or suramin was analyzed by flow cytometry (FCM). The results showed that that there was HSV-tk mRNA and protein expression in pDR2-tk plasmid transduced PC-3m cell. Combination of GCV with VP-16, VCR, 5-Fu or suramin led to an enhanced cellular killing effect, but with CDDP resulted in a reduced one and with MTX in an approximate one. FCM revealed that synergistic use of GCV and 5-fu or suramin resulted in a rather large proportion of apoptosis and necrosis with the apoptosis index being 36.38 % and 35.51%, and the proportion of necrosis being 33.05 % and 28.87 %, respectively. In conclusion, HSV-tk/CGV approach by addition of certain clinical available chemotherapeutic drugs brings on statistically significant enhanced cell killing over single-agent treatment.Our results highlight the potential for such new combination therapies for future treatments of HRPC.

To clone Uroplakin Ⅱ gene from Chinese transitional cell carcinoma (TCC) of bladder and construct its eukaryotic expression vector, the molecular cloning method was used to extract total RNA from a GⅢ/ T3N0M0 tissue sample of the bladder TCC patients. The primers were designed by Primer 5.0 software. Full length cDNA of Uroplakin Ⅱ gene was amplified by reverse transcription polymerase chain reaction (RT-PCR), assayed by nucleic acid sequencing and then inserted between Xba Ⅰ and HindⅢ restrictive sites of eukaryotic expression vector pcDNA3.0. The recombinant was assayed by restricted enzyme digestion. Under the induction of Lipofectamine 2000, the recombinant was transfected into Uroplakin Ⅱ negative bladder cancer cell line EJ. Cellular expression levels of Uroplakin Ⅱ were detected by RT-PCR. The nucleic acid sequencing results indicated that Chinese Uroplakin Ⅱ cDNA (555 bp) was successfully cloned. The BLAST analysis demonstrated that the cloned sequence is 100 % homologous with sequences reported overseas. The GenBank accession number AY455312 was also registered. The results of restricted enzyme digestion indicated that eukaryotic vector pcDNA-UP Ⅱ for Uroplakin Ⅱ was successfully constructed.After being transferred with pcDNA-UPⅡ for 72 h, cellular Uroplakin Ⅱ mRNA levels were significantly improved (P＜0.01). It is concluded that human Uroplakin Ⅱ gene was successfully cloned from Chinese TCC tissues, which provided a basis for further exploration of the roles of Uroplakin Ⅱ gene in TCC biological behaviors and potential strategies for targeted biological therapy of TCC.

To study the expression and significance of the serine protease Omi/HtrA2 in prostate cancer and benign prostatic hyperplasia. The expression of Omi/HtrA2 was assayed by means of immunohistochemical technique in 41 prostate cancer (Cap), 20 benign prostatic hyperplasia (BPH) and 10 normal prostate (NP) specimens. Omi/HtrA2 expression was positive in 30 (73.17%) prostate cancer specimens, and the positive rate of Omi/HtrA2 was lower in well differentiated than in poorly and moderately differentiated groups (P＜0.05). By contrast, the cells in normal prostate and benign prostatic hyperplasia groups showed no or weak expression of Omi/HtrA2.Prostate cancer cells in vivo may need Omi/HtrA2 expression for apoptosis, and that Omi/HtrA2expression might be involved in prostate cancer development.

To investigate the relationship of bcl-2, p53, proliferating cell nuclear antigen (PCNA) to cell proliferation, apoptosis and pathological parameters, the patterns of cell growth and turnover in renal cell carcinoma (RCC), formalin-fixed and paraffin-embedded tissue blocks from 34 patients with RCC were examined. Cell proliferation activity was detected by PCNA immunostaining and the proliferation index (PI) was expressed as a percentage of the PCNA-positive cells in the tumor cells. Apoptosis was detected by terminal deoxy- nucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and the apoptotic index (AI) was expressed as a percentage of the TUNEL-positive cells in the tumor cells. Expressions of bcl-2 and p53 were assessed immunohistochemically. Our results showed that the PI ranged from 6.0% to 24.0% (median 12.3%) and the AI from 2.0% to 8.0% (median 5.4%) in RCC. The expression of the bcl-2 protein was demonstrated in 15 cases (44.1%); the expression of the p53 protein, however, was seen in only 3 case. bcl-2 positivity was not associated with PI or AI or any pathological parameters. There were close associations between PI and tumor grade and stage, and a significant relationship between AI and the tumor grade of RCC, Our study suggests that bcl-2 positivity was not associated with PI or AI or any pathological parameters. There are close associations between PI and AI and tumor grade and stage of RCC. Active cell proliferation may be accompanied by frequent apoptosis in RCC.
Adult ; Aged ; Apoptosis ; physiology ; Carcinoma, Renal Cell ; metabolism ; pathology ; Cell Division ; Female ; Humans ; Kidney Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Tumor Cells, Cultured ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics