Objective To explore the application effect of advanced lung rehabilitation programs in patients un-dergoing high flow nasal cannula oxygen therapy(HFNC).Methods Convenience sampling method was used to se-lect 40 HFNC patients admitted to the respiratory department,ICU,coronary heart disease monitoring unit,and other departments of a tertiary A comprehensive hospital in Tianjin from January to June 2023 as the experimental group,and 40 HFNC patients admitted from June to December 2022 as the control group.Both groups of patients received HFNC treatment during hospitalization and continued at home,while the experimental group adopts the information-motivation-behavioral skills(IMB)model as the theoretical basis,implements advanced pulmonary rehabili-tation during the stable condition period,discharge preparation period,and home rehabilitation period,and imple-ments compliance management of home pulmonary rehabilitation through cloud follow-up,and the control group re-ceived routine lung rehabilitation and home follow-up.The degree of respiratory distress,6-minute walking distance,exercise self-efficacy,quality of life were compared between the 2 groups before intervention,2 months and 6 months after intervention,and compliance with home pulmonary rehabilitation was compared between the 2 groups 6 months after intervention.Results The repeated measures analysis of variance showed that there was an interaction effect between time and the 2 groups in terms of difficulty in breathing,6-minute walking distance,exercise self-ef-ficacy,and quality of life(P＜0.05).Simple effect analysis showed that after 2 and 6 months of intervention,the ex-perimental group performed better than the control group in the respiratory distress,6-minute walking distance,exer-cise self-efficacy,and quality of life(P＜0.05).Within 6 months of intervention,the compliance of home pulmonary rehabilitation in the experimental group was better than that in the control group,and the difference was statistically significant(P＜0.05).Conclusion The application of advanced pulmonary rehabilitation programme based on IMB can improve respiratory symptoms and improve exercise self-efficacy and adherence to pulmonary rehabilitation,en-hance activity endurance and improve quality of life.

To investigate the cellular target selectivity of small molecules targeting thioredoxin reductase 1, we reported the construction and functional research of a stable TrxR1 gene (encode thioredoxin reductase 1) knockout HCT-116 cell line. We designed and selected TrxR1 knockout sites according to the TrxR1 gene sequence and CRISPR/Cas9 target designing principles. SgRNA oligos based on the selected TrxR1 knockout sites were obtained. Next, we constructed knockout plasmid by cloning the sgRNA into the pCasCMV-Puro-U6 vector. After transfection of the plasmid into HCT-116 cells, TrxR1 knockout HCT-116 cells were selected using puromycin resistance. The TrxR1 knockout efficiency was identified and verified by DNA sequencing, immunoblotting, TRFS-green fluorescent probe, and cellular TrxR1 enzyme activity detection. Finally, the correlation between TrxR1 expression and cellular effects of drugs specifically targeting TrxR1 was investigated by CCK-8 assay. The results demonstrated that the knockout plasmid expressing the sgRNA effectively knocked-out TrxR1 gene within HCT-116 cells, and no expression of TrxR1 protein could be observed in stable TrxR1 knockout HCT-116 (HCT116-TrxR1-KO) cells. The TrxR1-targeting inhibitor auranofin did not show any inhibitory activity against either cellular TrxR1 enzyme activity or cell proliferation. Based on these results, we conclude that a stable TrxR1 gene knockout HCT-116 cell line was obtained through CRISPR/Cas9 techniques, which may facilitate investigating the role of TrxR1 in various diseases.
CRISPR-Cas Systems/genetics* ; Gene Editing ; Gene Knockout Techniques ; HCT116 Cells ; Humans ; RNA, Guide/metabolism*

Background: Several studies have shown that proton pump inhibitors (PPIs) can enhance the sensitivity of gastric cancer (GC) cells to chemotherapy and inhibit tumor proliferation and invasion. Aims: To investigate whether PPI could enhance chemosensitivity by inhibition of cell cycle-related genes in GC cells. Methods: Two human GC cell lines, AGS and HGC27 were treated with pantoprazole in different concentrations, and the cell viability was detected by CCK-8 assay. Transcriptome sequencing combined with KEGG enrichment analysis were used to determine the effect of PPI on cell cycle of GC cells, and the changes of cell cycle and its related genes were validated by flow cytometry, real-time PCR and Western blotting, respectively. Bioinformatics websites were employed to analyze the major differentially expressed cell cycle-related genes in GCs and their relationship with patients' prognosis. After transfection with FOXM1 plasmid or control plasmid, the inhibitory effect of PPI combined with cisplatin on GC cells was determined by CCK-8 assay. Results: PPI inhibited the proliferation of GC cells effectively in vitro. Transcriptome sequencing showed that the expression levels of G2/M phase-related genes, including FOXM1, PLK1, and AURKB were down-regulated in PPI-treated GC cells, and G2/M arrest was suggested by KEGG enrichment analysis. All these changes were proved by flow cytometry, real-time PCR and Western blotting. Bioinformatics analysis revealed that FOXM1, PLK1, and AURKB genes were highly expressed in GCs and correlated with a poor prognosis. The inhibitory effect of PPI combined with cisplatin on GC cells was superior to that of cisplatin alone, but could be partially reversed by overexpression of FOXM1. Conclusions: PPI treatment can induce G2/M arrest in GC cells by inhibiting cell cycle-related genes, and subsequently enhance the sensitivity of GC cells to chemotherapy.

In recent years,with the continuous expansion of graduates enrollment,the problem of graduates employment has attracted wide attention from all walks of life.As medical graduate students with favorable employment situation in the past,the pressure of employment now is increasing year by year,and the employment contradiction become more and more severe.In this paper,a survey of the graduates employment in the past five years in Medical University of Chongqing was conducted,and based on the analysis of the medical graduates employment situation and the characteristics of medical graduates training,six kinds of methods to improve employability for medical graduates were explored in order to achieve better employment of medical graduates.

Objective Diagnosis of contrast-enhanced ultrasound ( CEUS) for small focal liver lesions under the background of hepatocirrhosis has been a difficult problem in clinical practice .CEUS has significantly improved the accuracy of liver cancer diagno-sis, but little research has been done in minimal lesions .The article was to investigate the diagnostic value of CEUS in liver cirrhosis with small focal liver lesions (≤2 cm) . Methods 70 patients with cirrhosis and 76 small focal hepatic lesions who were admitted in our hospital from March 2015 to August 2015 were examined by conventional ultrasound and CEUS .Comparative analysis was made be-tween the results of tests and pathology to calculate the sensitivity , specificity and accuracy of CEUS and conventional ultrasound . Results The sensitivity , specificity and accuracy in diagnosing small hepatic cancer lesions complicated with cirrhosis by CEUS and conventional ultrasound were 47.6%、79.4%、61.8%and97.6%、97.1%、97.4%,respectively.CEUS was in high consistence with the results of pathology ( Kappa 0.947) and significantly higher than those of conventional ultrasound (Kappa 0.259). Conclusion CEUS has high application value in the differential diagnosis of small focal he-patic lesions complicated with cirrhosis .

AIM:To observe the effects of the combination of berberin and epirubicin on the cell cycle of T24 bladder cancer cells and the underlying mechanisms.METHODS:The cancer cells were exposed to epirubicin in the presence or absence of different concentrations of berberin.The viability of the cancer cells was determined by MTT assay.The cell cycle distribution was detected by flow cytometry, and the protein levels of cyclin D1, CDK2, CDK4, P21 and P27 were detected by Western blot.RESULTS:Berberine markedly enhanced the inhibitory effect of epirubicin on the viability of T24 cells and promoted epirubicin-induced cell cycle arrest at G0/G1 phase as compared with the negative control cells.Epirubicin increased the protein expression of P27 and P21, both of which were enhanced by treatment with berberin.In contrast, berberin exposure further decreased the protein expression of cyclin D1, CDK2 and CDK4 in epirubicin-treated T24 cells.CONCLUSION:Berberine significantly promotes epirubicin-induced G0 /G1 phase arrest in human bladder cancer cells by up-regulating P27 and P21 expression and inhibiting the expression of cyclin D1, CDK2 and CDK4.

Objective To investigate the therapeutic efficacy and safety of endoscopic resection of giant gastric stromal tumors without explicit evidence of metastases.Methods A total of 12 giant gastric stromal tumors with no evidence of metastases diagnosed by endoscopic ultrasound (EUS) and computed tomography (CT) scan were managed by endoscopic resection.Operation time,blood loss and the incidence rate of perforation were recorded respectively.The diagnoses of tissue specimens were made by pathological examination and immunohistochemistry.In order to assess local recurrence and distant metastases,endoscopy and endoscopic ultrasound follow-up examinations were performed routinely at 2,6 and 12 months,and the whole abdominal CT scan was also performed at 12 months after operation.Results Endoscopic resections were successfully performed in 10 of 12 cases (83.3％),among which,6 underwent endoscopic submucosal excavation (ESE) without unexpected perforation and 4 endoscopic full-thickness resection (EFR)with intentional perforation.The rate of intentional perforation was 33.3％ (4/12),and all the perforations could be sealed by endoscopic methods.The blood losses were all more than 100 ml,which could be controlled by argon plasma coagulation,electrocoagulation or hemostatic clips.In the 10 encapsulated tumors,8 could be smoothly removed from esophagus,whose long diameter of the minimum cross section was less than 3.5 cm,however,2 tumors whose diameters were larger than 3.5 cm were taken out after segmentation.In the 10 tissue samples,9 were confirmed as low risk GIST,1 larger than 5 cm was pathologically confirmed as high risk GIST.During 1-year follow-up,no local recurrence or peritoneal metastasis was found.2 tumors,larger than 5.0 cm,could not be removed by endoscopic methods due to uncontrolled bleeding.The rate of uncontrolled bleeding was 16.7％ (2/12).The patients were transferred to surgery,and pathologically confirmed as having high risk GIST.Conclusion For low-risk giant gastric stromal tumors whose diameters were less than 5cm without evidence of metastases,endoscopic resection is considered as a safe and effective procedure.Tumors with long diameter of the minimum cross section less than 3.5 cm are more suitable for endoscopic resection,which can be smoothly taken out through cardia.However,for high-risk GIST larger than 5.0 cm,the rate of uncontrolled bleeding is high,so endoscopic resection should be adopted with discretion.

Objective To study the expression and interactions of DNA methyltransferase 1 (DNMT1),enhancer of zeste homolog 2 (EZH2) and histone deacetylase 1 (HDAC1) in gastric cancer cell lines and tissue specimens.Methods The expression of DNMT1,EZH2 and HDAC1 was detected at mRNA and protein level in gastric cancer lines MKN28,SGC7901,BGC823,AGS,normal gastric epithelium cell line GES-1 and 10 pairs of fresh gastric cancer tissues and corresponding normal gastric tissues by real-time polymerase chain reaction and Western blot.Whether DNMT1,EZH2 and HDAC1 forming complex or not was detected by co-immunoprecipitation (Co-IP) in well-differentiated gastric cancer cell line MKN28,medium-differentiated gastric cancer cell line SGC7901,lowdifferentiated gastric cancer cell line BCG823,normal gastric epithelium cell line GES-1,mediumdifferentiated,medium to low-differentiated,low-differentiated gastric cancer tissues and corresponding normal gastric tissues.Results Compared with that of normal gastric epithelium cell and gastric tissue,the expression of DNMT1,EZH2 and HDAC1 in gastric cancer cell lines and gastric tissue was higher.The results of Co-IP indicated that DNMT1,EZH2 and HDAC1 formed complex in the high,medium,and poor differentiated gastric cancer cells and the medium,mediumlow,poor differentiated gastric cancer tissues,but not in normal gastric epithelium cell and tissue.Conclusion DNMT1,EZH2 and HDAC1 highly expressed in gastric cancer and there was interaction effects among them,which might be an important mechanism in the correlation between DNA methylation and histone modifications in gastric cancer.

Objective To investigate the clinical,pathologic and endoscopic ultrasound characteristics of pancreatic neuroendocrine tumors (PNETs).Methods Clinical data of 24 consecutive patients of PNETs who were admitted between January 2002 and January 2011 were reviewed.Results Among these 24 patients,19 were diagnosed to have insulinomas,1 was malignant insulinoma,2 were gastrinomas,and 2 were glucagonomas.Eighteen (75％) cases of PNETs were functional PNETs,and 6 (25％) were nonfunctional PNETs.The mean age of the patients was (42 ± 14) years old ranging from 19 ～ 64 years old,and the percentage of male patient was 33.3％.The main symptoms of insulinomas were intermittent abdominal pain or discomfort,and the main manifestations of glucagonomas were weight loss,skin migratory erythema; and the main symptoms of gastrinomas were diarrhea,vomiting with large amounts of water-like liquid.The detection rate of CT was 86.7％ (13/15),and the detection rate of EUS was 100％ (15/15),but PET-CT detected only 40％ of tumors (2/5).The endoscopic ultrasound characteristics of PNETs were circular or oval hypoechoic mass,and the volume was small with clear boundary and homogeneous echo.There was no enlarged lymph node,and liquid was detected in big tumor.Twenty-two patients received operation and 2 patients did not.PNETs expressed CgA and Syn protein.All patients of PNETs were alive with 7 to 80 months follow-up.Conclusions The clinical characteristics of PNETs were unique.EUS has a high accuracy for detecting and localizing PNETs.The surgical method is similar to that of pancreatic cancer,and the prognosis is relatively good.

ObjectiveTo investigate the clinicopathologic features,therapy,and prognosis of primary small cell carcinoma of the pancreas.MethodsDatabases including Chinese Journal Full-text Database,VIP Database for Chinese Technical Periodicals,Medline/Pubmed,and OVID were searched electronically up to April 2012.A systematic review was performed together with one case in our hospital.ResultsTwenty-eight articles fulfilling the criteria consisting of 46 patients with pathologically confirmed diagnosis of primary small cell carcinoma of the pancreas were studied,together with 1 patient in our Drum Tower Hospital,finally 47patients were included.The results of this systematic review showed:( 1 ) Primary small cell carcinoma of the pancreas was more common in men with a median age of 62.The most common clinical presentations were abdominal pain,jaundice and weight loss.Para-neoplastic syndrome was rarely observed.(2)Most cases were found to have abnormally elevated serum levels of neuron-specific enolase.CT displayed heterogeneous,and marked enhancing masses in most cases.The conclusive diagnosis depended on histological confirmation.(3)63.8％ of the cases were found to be associated with metastasis at the time of diagnosis.The overall median survival time was 28 weeks.(4) There was no consensus on the treatment of primary small cell carcinoma of the pancreas. Chemotherapy was currently considered as the treatment of choice among the systematic management for these patients.ConclusionsPrimary small cell carcinoma of the pancreas was a rare and aggressive neuroendocrine tumor with a poor prognosis.