ObjectiveTo investigate the change in the activity of hepatitis B virus （HBV）-specific CD8⁺ T cells after pegylated interferon-α-2b （PEG-IFN-α-2b） therapy in HBeAg-negative patients with chronic HBV infection. MethodsA total of 53 HBeAg-negative patients with chronic HBV infection who attended The First Affiliated Hospital of Xinxiang Medical University and Tangdu Hospital of Air Force Mdical University from April 2020 to June 2022 were enrolled and treated with PEG-IFN-α-2b （180 μg/week， subcutaneous injection） antiviral therapy. The study endpoint was HBsAg clearance （course of treatment<48 weeks） or 48 weeks （course of treatment≥48 weeks）. Peripheral blood mononuclear cells were isolated at baseline and study endpoint， and peripheral blood T cell counts were measured. Enzyme-linked immunospot assay was used to measure the frequency of HBV-specific CD8⁺ T cells secreting perforin， granzyme B， and interferon-γ. A total of 17 HLA-A^*02-restricted patients were selected， and CD8⁺ T cells were purified to establish direct- and indirect-contact co-culture systems for HBV-specific CD8⁺ T cells and HepG2.2.15 cells. The level of lactate dehydrogenase in supernatant was measured to calculate the mortality rate of HepG2.2.15 cells， and the levels of HBV DNA， cytotoxic molecules， and cytokines in supernatant were also measured. Flow cytometry was used to measure the expression of apoptosis ligands， and the cytotoxicity of HBV-specific CD8⁺ T cells was evaluated. The independent samples t-test or the paired t-test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test or the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups. ResultsThe HBsAg clearance rate at study endpoint was 30.19% （16/53）. There were no significant differences in peripheral blood T cell counts （CD3⁺， CD4⁺， and CD8⁺ T cells） between baseline and study endpoint （P>0.05）. At study endpoint， there was a significant increase in the frequency of HBV-specific CD8⁺ T cells secreting perforin， granzyme B， and interferon-γ （U=177.50， t=11.90， U=186.50， all P<0.001）， and the patients with HBsAg clearance had a significantly higher frequency of such HBV-specific CD8⁺ T cells than those without HBsAg clearance （U=120.50， t=2.73， U=121.50， all P<0.01）. In the direct- and indirect-contact co-culture systems at study endpoint， HBV-specific CD8⁺ T cells induced a significant reduction in HBV DNA in the supernatant of HepG2.2.15 cells （all P<0.001） and significant increases in the secretion of interferon-γ and tumor necrosis factor-α （all P<0.05）； in the direct-contact co-culture system， HBV-specific CD8⁺ T cells induced significant increases in the mortality rate of HepG2.2.15 cells （13.62%±3.27% vs 11.39%±2.40%， t=2.27， P=0.030） and the secretion of perforin and granzyme B （t=72.50， U=52.50， both P<0.05）. In the direct- and indirect-contact co-culture systems， compared with HBV-specific CD8⁺ T cells from the patients without HBsAg clearance， the HBV-specific CD8⁺ T cells from patients with HBsAg clearance had a significantly greater reduction in HBV DNA （P<0.05） and significant increases in the secretion of interferon-γ and tumor necrosis factor-α （P<0.05）. ConclusionPEG-IFN-‍α‍-2b therapy can help to achieve a relatively high HBsAg clearance rate in HBeAg-negative patients with chronic HBV infection， and the activity of HBV-specific CD8⁺ T cells is significantly enhanced， which is closely associated with HBsAg clearance.

Objective:To investigate the clinical efficacy, quality of life, safety, and medical expense of second-line treatment with ametinib versus first-line treatment with oxitinib in patients with advanced non-small cell lung cancer (NSCLC) harboring epidermal growth factor receptor (EGFR) mutations. Methods:This is a prospective clinical research study. A total of 72 patients with NSCLC who received second-line treatment with ametinib at the Affiliated Hospital of Jining Medical University from July 2020 to April 2022 were included in the amitinib group. Sixty-six patients with advanced NSCLC harboring EGFR mutations who underwent first-line treatment with ositinib were included in the ositinib group (one patient dropped out of the study, and sixty-five patients were included in the final analysis). There was no statistically significant difference in general information between the two groups. The ositinib group received ositinib treatment, while the amitinib group received amitinib treatment. After three courses of treatment, the short-term effectiveness was evaluated in both groups. The quality of life was compared before and after three courses of treatment in each group. Adverse reactions and average per-day hospital cost were compared between the two groups.Results:The disease control rate in the amitinib and oxitinib groups were 91.67% (66/72) and 90.77% (59/65), respectively, while the overall response rate was 43.06% (31/72) and 41.54% (27/65), respectively. The disease control rate and overall response rate did not differ significantly between the two groups (both P > 0.05). After treatment, the scores for physical well-being [(21.05 ± 4.18) points vs. (19.16 ± 3.95) points, t = 4.05], social/family well-being [(19.38 ± 2.65) points vs. (17.26 ± 2.28) points, t = 3.11], emotional well-being [(18.83 ± 3.07) points vs. (17.00 ± 2.86) points, t = 3.20], functional well-being [(22.02 ± 3.83) points vs. (20.14 ± 2.98) points, t = 3.83], and additional attention [(26.11 ± 5.00) points vs. (24.33 ± 4.30) points, t = 3.05] in the amitinib group were significantly higher than those in the oxitinib group (all P < 0.05). There was no significant difference in incidence rate of grade 3-4 adverse reactions between amitinib and oxitinib groups [6.94% (5/72) vs. 10.77% (7/65), P > 0.05]. Average per-day hospital cost in the amitinib group was significantly lower than that in the oxitinib group ( t = 4.83, P < 0.05). Conclusion:Second-line treatment with ametinib for advanced NSCLC harboring EGFR mutations can significantly enhance the quality of life and offer comparable short-term efficacy and safety to first-line treatment with oxitinib. Advantageously, its medical cost is relatively lower.

Objective To analyze the values of serum tumor necrosis factor-α stimulated gene 6 protein (TSG-6) and type ⅩⅥ collagen (col-16) in predicting the prognosis of patients with ulcerative colitis (UC). Methods A total of 160 patients with UC were enrolled as UC group, another 160 volunteers were selected as control group, and the levels of serum TSG-6 and col-16 were compared between the two groups. According to the modified Mayo scoring criteria, patients in the UC group were divided into mild group (n=63), moderate group (n=52), and severe group (n=45), and the levels of serum TSG-6 and col-16 were compared among the three groups. Based on the colonoscopy follow-up results, patients in the UC group were divided into good prognosis group (n=121) and poor prognosis group (n=39), and the levels of serum TSG-6 and col-16 were compared between the two groups. Spearman correlation analysis was used to explore the correlations of serum TSG-6 and col-16 with the modified Mayo score. Logistic regression analysis was used to investigate the impact of serum TSG-6 and col-16 on the prognosis of patients in the UC group. Receiver operating characteristic (ROC) curve was used to assess the values of serum TSG-6 and col-16 in predicting the prognosis of patients in the UC group. Results The levels of serum TSG-6 and col-16 in the UC group were significantly higher than those in the control group (P < 0.01). The levels of serum TSG-6 and col-16 in the moderate and severe groups were significantly higher than those in the mild group, and the levels of serum TSG-6 and col-16 in the severe group were significantly higher than those in the moderate group (P < 0.01). The levels of serum TSG-6 and col-16 in the poor prognosis group were significantly higher than those in the good prognosis group (P < 0.01). Spearman correlation analysis showedthat serum TSG-6 and col-16 were positively correlated with the modified Mayo score (r=0.691, P < 0.05; r=0.635, P < 0.05). Logistic analysis results showed that serum TSG-6 and col-16 were risk factors for poor prognosis (P < 0.001). ROC curve analysis showed that the areas under the curve for predicting poor prognosis in patients with UC were 0.773 and 0.765 for serum TSG-6 and col-16, respectively. Conclusion The levels of serum TSG-6 and col-16 are abnormally elevated in patients with UC, and there is a certain correlation of the two indexes with the severity and prognosis of disease, and the two indexes can be used as predictors of poor prognosis.

Objective To investigate the effects of tegafur, gimeracil and oteracil potassium combined with oxaliplatin on gastric motility-related hormones, matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in elderly patients with gastric cancer. Methods A total of 128 elderly patients with gastric cancer were selected as the study subjects and randomly divided into control group (n=64) and observation group (n=64). The control group was treated with tegafur, gimeracil and oteracil potassium, while the observation group received tegafur, gimeracil and oteracil potassium combined with oxaliplatin. Gastric motility-related hormones [motilin (MTL) and vasoactive intestinal peptide (VIP)], MMP-2, MMP-9, tumor markers [carcinoembryonic antigen (CEA), carbohydrate antigen 125 (CA125) and carbohydrate antigen 19-9 (CA19-9)], clinical efficacy and occurrence of adverse reactions were observed in both groups. The correlations of gastric motility-related hormones with the levels of MMP-2, MMP-9 and tumor markers were analyzed. Results After treatment, MTL levels in both groups were significantly lower, while VIP levels were significantly higher compared to before treatment. Additionally, the observation group had significantly lower MTL and higher VIP levels compared to the control group (P < 0.05). After treatment, the levels of MMP-2, MMP-9, CEA, CA125 and CA19-9 were significantly reduced in both groups compared to pretreatment levels, with the observation group showing significantly lower levels than the control group (P < 0.05). The total effective rate in the observation group was 70.31%, which was significantly higher than the 53.13% in the control group (P < 0.05). There was no statistically significant difference in the incidence of adverse reactions between the two groups (P>0.05). A negative correlation was observed between MTL and MMP-2, MMP-9, CEA, CA125 as well as CA19-9 (P < 0.05), while VIP showed a positive correlation with MMP-2, MMP-9, CEA, CA125 and CA19-9 (P < 0.05). Conclusion The combination of tegafur, gimeracil and oteracil potassium and oxaliplatin is effective and safe for the treatment of elderly patients with gastric cancer. MTL, VIP, MMP-2, MMP-9, CEA, CA125 and CA19-9 are interrelated in the development of gastric cancer in elderly patients.

Objective:To understand the cognition and training status of basic life support among medical staff in Linfen, Shanxi Province, and to provide reference for the development of targeted training strategies and programs.Methods:A questionnaire survey was conducted among medical staff in 12 county hospitals in Linfen, Shanxi Province by convenience sampling method. The survey included the general characteristics of departments and medical staff, previous basic life support training assessment and cognitive status.Results:A total of 839 medical staff were included, 756 (90.1%) completed the survey, 183 (24.2%) were doctors and 573 (75.8%) were nurses. Most personnel lacked awareness of environmental safety, emergency response system start-up, adequate compression, airway management, and electrical defibrillation.Conclusions:The cognitive status of basic life support of medical staff in Linfen county is not optimistic. It is necessary to construct an applicable precision training course and retraining assessment system to improve the cognitive level and practical operation ability.

Objective To compare the detection effects of Legionella pneumophila in water samples from public places by the Legiolert enzyme-substrate method and the conventional cultivation. Methods The Legiolert enzyme-substrate method and the conventional cultivation method were used to detect Legionella pneumophila in cooling water and shower water samples collected in public places. Isolated strains were verified and serotyped. Results A total of 68 samples were collected and tested. The positive rate of the conventional cultivation and the Legiolert enzyme-substrate method were 5.88%（4/68）and 35.29%（24/68）, respectively, with a significant difference （χ²=16.41，P<0.01）. The coincidence rate of the two methods was 64.71% (44/68), and the difference of the detection effects of the two methods was statistically significant (χ²=16.41，P=0.000). A total of 25 strains of Legionella pneumophila were isolated，and the serum types were mainly LP1（14/25）. Conclusion The Legiolert enzyme-substrate method represente a higher detection rate of Legionella pneumophila in water samples from public places than the conventional cultivation.

Objective To find out the current situation and related factors of perioperative pain management for the patients with fracture among the top three hospitals in our region.Methods A general information questionnaire was made,to survey the the status and the influencing factors of top hospital nursing work of fractures patients with perioperative pain management.The researchers label the questionnaire with the method of digital label,and the data collected are kept secret,and the statistical method is used to evaluate it.Finally,the conclusion is drawn.Results The working life of the nursing staff,nursing education,nursing staff after underwent fracture perioperative pain knowledge of continuing education is the mainly factors to affect patients surfing with fracture,perioperative pain management cycle of practice the three most important factors.Conclusion Three tertiary hospitals in the area fracture nursing of pain management need to be strengthened.we strengthen fractures patients with perioperative pain management cycle practice by training of clinical nursing staff.

Objective To investigate the clinical and pathological characteristics, diagnosis and differential diagnosis of uterine intravenous leiomyoma (IVL) , so as to improve the detection of the disease.Methods Five cases of IVL were retrospected, who were treated and diagnosed by clinical gross examination,morphologic features and immunohistochemical staining in Tumor Hospital of Anyang from 2011 to 2015.Results Symptoms of IVL were usually nonspecific, most patients experienced hypermenorrhea, abdominal pain or distension, and pelvic mass.Typical pathological changes were intravenous visible grey weblike or worm-formed nodules, which could be pulled out.Microscopically, the tumors were composed of mild spindle-shaped cells with rare mitotic figure and closely related to blood vessels.The tumors were positive for smooth muscle derived marker, progesterone receptor, estrogen receptor, and negative for S-100, HMB45 and CD34.However CD34 can displayed vascular endothelial cells around the tumor.Conclusion Uterine IVL is a benign tumor with malignant biological behavior, which has different clinical pathological features of leiomyoma ordinary.Surgery is the main treatment method.A satisfactory therapeutic effect can be achieved by selecting the appropriate extent of surgery.Long term follow up of iVL patients after operation is important.

Objective To establish a simple and effective method for isolation and culture of mouse adipose-derived stem cells (mASCs)in vitro,in order to provide the sufficient sources of seed cells for the research of mesenchymal stem cells.Methods The mouse inguinal fat tissues were isolated in vitro and performed a digestion with 0.1% collagenase type NB4,then adipose-derived stem cells(ASCs)were seeded and adhered to the culture dishes in low glucose DMEM containing 10% fetal calf serum.The cellu-lar morphology,in vitro proliferation capacity,multidifferentiation potential and immunophenotype were assessed.Results The mASCs showed good cell morphology,extremely strong proliferation capacity and potential of adipogenesis,osteogenesis and chon-drogenesis via in vitro three-dimensional induction.The cellular surface antigen phenotype was consistent with that reported by lit-erature,and the expression of CD34 and CD105 was positive,Sca-1 was highly expressed,CD45 and SSEA-1 were not expressed. Conclusion Using the experimental methods in this research can culture the high purity of mASCs with the excellent stem cell properties and extremely strong proliferative ability.

Aim To study the interference role of Bcl-2 siRNA on HL-60 cells. Methods Bcl-2 siRNA was synthesized in vitro transcription with silencer siRNA construction kit. The synthesized siRNA was transfected into HL-60 cells with lipid siPORT transfection. Forty-eight hours after the transfection, we used MTT and immunofluorescence to detect cell proliferation and apoptosis,and used RT-PCR and immunofluorescence to detect the level of Bcl-2 mRNA and Bcl-2 protein expression. Result Bcl-2 siRNA reduced the level of Bcl-2 mRNA and Bcl-2 protein expression in HL-60 cells and induced cell apoptosis. There was no difference on the effect of other groups compared with the control. Effective Bcl-2 siRNA specifically degraded Bcl-2 expression in the levels of mRNA and protein and induced HL-60 cells apoptosis.Conclusion These results indicate that siRNA is a highly specific tool for targeted gene knockdown. siRNA-mediated gene silencing is a reliable approach for large-scale screening of gene function and drug target validation.