BACKGROUND:Small diameter artificial vessels are urgently needed to treat coronary artery and peripheral artery diseases in clinical practice.At present,vascular tissue engineering has become the main method for preparing small diameter artificial vessels.Selecting suitable biomaterials and cell sources is the key factor for successful construction of small diameter tissue engineered vessels. OBJECTIVE:To observe the effect of four kinds of electrospinning membrane materials on proliferation,adhesion and differentiation of bone marrow mesenchymal stem cells into vascular smooth muscle cells. METHODS:Bone marrow mesenchymal stem cells were isolated and extracted from SD rats.The bone marrow mesenchymal stem cells were inoculated separately on polycaprolactone(PCL),polycaprolactone-hyaluronic acid(PCL-HA),polycaprolactone-silk-filament proteins(PCL-SF),and polycaprolactone-gelatin(PCL-GEL)electrospinning membrane materials.After 1,3,and 7 days of culture,the cell arrangement on the material was observed under scanning electron microscope.The proliferation and adhesion of the material were observed by phalloidin staining.The mRNA expressions of CD90,Meflin,and transforming growth factor β were detected by qRT-PCR.After 7 days of induced differentiation into vascular smooth muscle cells,the mRNA expression ofɑ-smooth muscle actin on the material was detected by qRT-PCR. RESULTS AND CONCLUSION:(1)Bone marrow mesenchymal stem cells were arranged along the fibers of the four kinds of electrospinning membranes under scanning electron microscopy.(2)Phalloidin staining showed the regular distribution of bone marrow mesenchymal stem cells on the four kinds of electrospinning membranes and parallel distribution along the fiber direction.Moreover,PCL-HA,PCL-SF,and PCL-GEL electrospinning membranes were more conducive to the proliferation and adhesion of bone marrow mesenchymal stem cells than PCL electrospinning membranes.Compared with PCL-HA and PCL-GEL electrospinning membranes,PCL-SF electrospinning membranes were more conducive to the proliferation and adhesion of bone marrow mesenchymal stem cells.(3)qRT-PCR showed that the four kinds of electrospun membrane materials could maintain the mRNA expression of CD90 and Meflin in bone marrow mesenchymal stem cells,but there was no significant difference between groups(P>0.05).The mRNA expression of transforming growth factor β in PCL-HA,PCL-SF,and PCL-GEL groups was higher than that in PCL group on days 1 and 7(P<0.05),and the mRNA expression of transforming growth factor β in PCL-SF group was higher than that in the other three groups on days 3 and 7(P<0.05).The mRNA expression of transforming growth factor β in PCL-HA group was higher than that in PCL-GEL group on day 7(P<0.05).(4)qRT-PCR showed that the mRNA expression of ɑ-smooth muscle actin in PCL-SF group was higher than that in the other three groups(P<0.05),and that in PCL-HA group was higher than that in PCL group(P<0.05).(5)The results indicate that compared with PCL,PCL-HA and PCL-GEL electrospinning membranes,PCL-SF electrospinning membranes combined with bone marrow mesenchymal stem cells are more suitable for the preparation of small diameter tissue engineered vessels.

OBJECTIVE To assess the cost-effectiveness of durvalumab combined with chemotherapy as a first-line treatment for advanced biliary tract cancer from the perspective of the Chinese healthcare system. METHODS Using data from the TOPAZ-1 clinical trial， a three-state Markov model comprising progression-free survival （PFS）， progressive disease （PD） and death was developed， with a cycle length of 21 days and a 10-year time horizon. Patients in the observation group received durvalumab in combination with gemcitabine and cisplatin， whereas those in the control group received placebo plus the same chemotherapy regimen. The evaluation indexes were quality-adjusted life year （QALY） and the incremental cost-effectiveness ratio （ICER）. The willingness-to-pay （WTP） threshold was set at three times the 2024 Chinese per capita gross domestic product （GDP） （287 247 yuan/QALY）. The sensitivity analyses， along with scenario analyses， were performed. RESULTS In the base-case analysis， the ICER of observation group compared to control group was 1 166 344.46 yuan/QALY， far exceeding the WTP threshold， indicating that the regimen was not cost-effective. One-way sensitivity analysis identified the PD state utility， discount rate， cost of durvalumab， and PFS state utility as the main drivers of ICER variation. Probabilistic sensitivity analysis showed that， at the above WTP threshold， the probability of the acceeptance of this regimen was 0， further supporting the robustness of the base-case findings. In the scenario analysis， inclusion of a patient assistance program reduced the ICER to 235 885.16 yuan/ QALY， below the above WTP threshold， suggesting cost-effectiveness under this assistance program. However， when applying a regional WTP threshold set at three times the per capita GDP （158 475 yuan/QALY） of Gansu Province （the province with the lowest GDP in China in 2024）， the ICER remained above the threshold， indicating that the regimen was not cost-effective at the regional level. CONCLUSIONS At current pricing， durvalumab plus chemotherapy as a first-line treatment for advanced biliary tract cancer is not cost-effective in China. Although the introduction of a patient assistance program can substantially reduce the ICER and achieve cost-effectiveness at a WTP threshold set at three times the 2024 per capita GDP of China， due to limited affordability in low-income areas， the program remains not cost-effective.

Objective: To investigate the expressions of GPR15 and FOXP3 in colorectal carcinoma(CRC) tissues and their clinical prognostic values. Methods : A total of 132 patients with CRC underwent radical surgery were collected. The control group selected the normal mucosal tissues more than 5 cm away from the edge of the cancer focus. Immunohistochemistry(Envision two-step method) was used to detect the expression levels of GPR15 and FOXP3 in CRC and adjacent tissues, and analyze their relationships with clinicopathological factors of colorectal cancer. Kaplan-Meier method was used to draw the survival curve to analyze the correlation between the expressions of GPR15 and FOXP3 and the survival prognosis of patients with CRC. The factors influencing prognosis of patients with colorectal cancer were analyzed by Cox regression. Results : The immunohistochemistry showed that the expression levels of GPR15 and FOXP3 in CRC were significantly higher than those in normal colorectal mucosal tissues(P<0.05). The expression of GPR15 in CRC tissues was correlated with location, nerve invasion and TNM stage; FOXP3 expression was correlated with sex(P<0.05).Both expressions were not significantly correlated with the clinicopathologic features of age, tumor size, differentiation degree, tissue type, depth of invasion, tumor budding, vascular invasion and lymph node metastasis. Correlation analysis showed that there was no significant correlation between GPR15 and FOXP3 expression(Kappa=-0.019,P>0.05). The survival prognosis of GPR15 positive group was significantly worse than that of negative group(log-rank: χ2=4.3,P=0.039);while the survival prognosis of FOXP3 positive group was significantly better than that of negative group(log-rank: χ2=7.3,P=0.007).Age ≤55 years, positive GPR15 and negative FOXP3 were independent risk factors for poor prognosis in patients with CRC(P<0.05). Conclusion The expression levels of GPR15 and FOXP3 in CRC are significantly higher than those in paracancer tissues, GPR15 and FOXP3 are expected to become new tumor markers for early screening, accurate treatment and prognosis assessment of CRC.

Objective To explore the safety and efficacy of laparoscopic left renal vein stent implantation combined with microsurgical varicocelectomy in the treatment of nutcracker syndrome complicated with varicocele.Methods A retrospective analysis was conducted on the data of 50 patients with nutcracker syndrome complicated with varicocele admitted to our hospital during Nov.2018 and Jul.2023.All patients underwent laparoscopic left renal vein stent implantation combined with microsurgical varicocelectomy.Results Altogether 47 patients were followed up after operation,with a mean age of(21.45±3.84)years.All operations were successful,and gross hematuria,proteinuria,abdominal pain and other symptoms relieved within 3-7 days after surgery.Postoperative follow-up showed that no patients had serious complications or recurrence of clinical symptoms.Left renal vein ultrasound and CT showed that the blood flow of the left renal vein was restored,and the stent position was stable.Compared with those before surgery,the angle between the abdominal aorta and the superior mesenteric artery[(44.91±6.59)° vs.(22.58±6.61)°]and the diameter of the left renal vein at the angle[(6.03±0.99)mm vs.(1.87±0.46)mm]were significantly increased,and the blood flow velocity of the left renal vein at the angle[(48.77±14.79)cm/s vs.(102.53±41.15)cm/s]was significantly decreased,with statistically significant differences(P＜0.001).Scrotal ultrasound confirmed that there was no recurrence of varicocele 6 months after surgery,and the diameter of the spermatic vein was significantly reduced,with statistically significant differences(P＜0.001).Semen analysis showed that the sperm concentration[(60.27±48.45)×106/mL vs.(36.57±36.67)×106/mL,P=0.032]and percentage of rapid forward movement of sperm were significantly increased[(22.54±10.70)vs.(15.01±10.77),P=0.005],with statistically significant differences.The increase value of semen concentration[(23.7±41.48)×106/mL vs.(6.12±4.97)×106/mL,P=0.016)]increased after combined operation compared with left renal venous stenting alone,and there was no significant difference in the diameter of spermatic vein,inner diameter of renal vein and flow rate between the two surgical methods(P＞0.05).Conclusion Laparoscopic left renal vein stent implantation combined with microsurgical varicocelectomy is a minimally invasive,safe and effective method for the treatment of nutcracker syndrome complicated with varicocele.

Objective:To investigate the feasibility of oblique overlap anastomosis plus single flap valvuloplasty (OSF) for reconstruction after laparoscopic proximal gastrectomy.Methods:The C-shaped seromuscular flap (2.5 × 3.5 cm), which was 2 cm from the top of the remnant stomach, was extracorporeally created on the anterior wall of the remnant stomach. The stomach was opened approximately 1.0 cm above the lower edge of the mucosal flap. Four supporting stitches were sutured around the hole and the right wall of the esophageal stump was incised with a support suture to prevent the linear stapler from entering the submucosa. Liner stapler was inclined to the left side of the esophagus at an angle of about 30 degrees to the longitudinal axis of the esophagus for oblique anastomosis between the dorsal side of the esophagus and the anterior stomach and then esophagogastrostomy was performed with the length of anastomosis was 4 cm. Entry hole was suture with 3 stitches and then the linear stapler was used for closing the entry hole. Finally, the seromuscular flap was closed using barbed sutures.Results:Clinical data of 11 patients with Siewert type II esophagogastric junction adenocarcinoma who underwent radical gastrectomy and reconstruction by OSF between January 2022 and May 2023 were retrospectively collected. There were 7 males and 4 females. The average age was (69.9±7.8) years, the BMI was (21.7±7.2) kg/m 2 and the tumor size was (2.1±0.6) cm. OSF reconstruction was successfully completed in all 11 patients. The median operative time was 275 (270-428) minutes, the time for OSF reconstruction was 112 (80-140) minutes, and the blood loss was 50 (20-400) ml. The pathological stage was 0-I in 7 cases and II-III in 4 cases. The patients were fed on the 4th day (4-7 days) and discharged from hospital on the 7th day (6-9 days) after surgery. No patient had gastroesophageal reflux symptoms of grade B and above, and no patient took anti-reflux medicine. Conclusions:OSF is a safe and feasible treatment for Siewert type II esophagogastric junction adenocarcinoma.

Objective:To investigate the feasibility of oblique overlap anastomosis plus single flap valvuloplasty (OSF) for reconstruction after laparoscopic proximal gastrectomy.Methods:The C-shaped seromuscular flap (2.5 × 3.5 cm), which was 2 cm from the top of the remnant stomach, was extracorporeally created on the anterior wall of the remnant stomach. The stomach was opened approximately 1.0 cm above the lower edge of the mucosal flap. Four supporting stitches were sutured around the hole and the right wall of the esophageal stump was incised with a support suture to prevent the linear stapler from entering the submucosa. Liner stapler was inclined to the left side of the esophagus at an angle of about 30 degrees to the longitudinal axis of the esophagus for oblique anastomosis between the dorsal side of the esophagus and the anterior stomach and then esophagogastrostomy was performed with the length of anastomosis was 4 cm. Entry hole was suture with 3 stitches and then the linear stapler was used for closing the entry hole. Finally, the seromuscular flap was closed using barbed sutures.Results:Clinical data of 11 patients with Siewert type II esophagogastric junction adenocarcinoma who underwent radical gastrectomy and reconstruction by OSF between January 2022 and May 2023 were retrospectively collected. There were 7 males and 4 females. The average age was (69.9±7.8) years, the BMI was (21.7±7.2) kg/m 2 and the tumor size was (2.1±0.6) cm. OSF reconstruction was successfully completed in all 11 patients. The median operative time was 275 (270-428) minutes, the time for OSF reconstruction was 112 (80-140) minutes, and the blood loss was 50 (20-400) ml. The pathological stage was 0-I in 7 cases and II-III in 4 cases. The patients were fed on the 4th day (4-7 days) and discharged from hospital on the 7th day (6-9 days) after surgery. No patient had gastroesophageal reflux symptoms of grade B and above, and no patient took anti-reflux medicine. Conclusions:OSF is a safe and feasible treatment for Siewert type II esophagogastric junction adenocarcinoma.

Objective:To explore the mechanism of peritoneal dialysis solution (PDS)-induced peritoneal microinflammation through activation of ceramide (CER) in peritoneal dialysis model mice.Methods:Thirty 5-week-old male C57BL/6 mice weighing about 22 g were used to set up peritoneal dialysis models, and then were randomly divided into 4 groups: sham operation group (1.5 ml sterilized water, n=7), high glucose-PDS group (1.5 ml 4.25% PDS, n=8), high glucose-PDS+ acid sphingomyelinase (ASMase) inhibitor desipramine (DES) group (1.5 ml sterilized water+10 mg/kg DES, n=8), high glucose-PDS+Src kinase inhibitor PP2 group (1.5 ml sterilized water +1 mg/kg PP2, n=7), with intraperitoneal injection once a day. After 28 days, the mice were sacrificed to retain peritoneal tissues. HE staining and Masson staining were used to observe the histological changes of peritoneum. Immunohistochemistry was used to detect the Toll-like receptor 4 (TLR4) and macrophages. High performance liquid chromatography, liquid chromatography/mass spectrometry and immunofluorescence were used to detect the expression of ASMase and CER. Real-time quantitative PCR was used to detect the mRNA levels of c-Src, p-Src, interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Western blotting was used to detect the protein levels of c-Src, and p-Src. Enzyme-linked immunosorbent assay was used to detect the serum C reactive protein (CRP), IL-6 and TNF-α. Results:(1) High glucose-PDS led to peritoneal hyperplasia, collagen deposition and fibrosis in the peritoneal dialysis mice, indicating successful modeling. Compared with high glucose-PDS group, peritoneal hyperplasia, collagen deposition and fibrosis of mice treated with DES and PP2 were significantly improved (all P<0.05). (2) Compared with sham operation group, ASMase activation and CER level of peritoneal tissues were significantly higher in high glucose-PDS group, and DES could significantly inhibit activated ASMase and increased CER expression caused by high glucose-PDS (both P<0.05). PP2 had no significant effect on ASMase activation and CER level (both P>0.05). (3) Compared with sham operation group, there were more TLR4 and macrophage positive staining cells in peritoneal tissues in high glucose-PDS group, and the mRNA expression levels of IL-6 and TNF-α in peritoneal tissues and serum CRP, IL-6 and TNF-α were higher (all P<0.05). DES and PP2 could significantly inhibit the increased TLR4, macrophages and related inflammatory factors induced by high glucose-PDS (all P<0.05). (4) Compared with sham operation group, c-Src and p-Src mRNA and protein expression levels of peritoneal tissues in high glucose-PDS group were significantly higher (all P<0.05). PP2 significantly inhibited the increased p-Src mRNA and protein levels caused by high glucose-PDS (both P<0.05), but had no significant effect on the mRNA and protein expression levels of c-Src (both P>0.05). DES had no significant effect on the mRNA and protein expression levels of c-Src and p-Src (all P>0.05). Conclusions:High glucose-PDS may enhance the expression of CER through stimulating the activity of ASMase, phosphorylate Src, activate TLR4 and induce inflammatory damage of peritoneum in peritoneal dialysis model mice.

Objective:To analyze any effect of combining hyperbaric oxygen with qingkailing on the metabolic biomarkers of consciousness disorder (CD) caused by brain trauma using metabolomics technology. Also, to screen out diagnostic markers for clinical treatment and prognosis.Methods:Thirty-six patients with a CD caused by brain trauma formed the observation group, while 40 healthy gender-, age- and ethnicity-matched individuals were the control group. Both groups were given routine supportive treatment, while the observation group additionally received hyperbaric oxygen and oral qingkailing medication. Before and after 14 days consciousness disturbance was evaluated using the Glasgow Coma Scale (GCS). Venous blood was collected for metabolomic analysis using liquid chromatography linked to a mass spectrometer to screen out specific metabolic biomarkers. Metabolic markers associated with the disease and of diagnostic significance were thus identified.Results:After the treatment the average GCS score of the observation group had improved significantly and the degree of coma was significantly relieved. Twenty-one metabolic markers were found to be significant, with creatine, proline, uric acid, acetyl-L-carnitine, histidine, proline leucine, tryptophan and 9E-octagenoic acid potentially of high diagnostic value. After the treatment, all of those markers came close to the levels observed in the healthy control group.Conclusions:Proline, leucine, 9E-octagenoleic acid, uric acid and acetyl-L-carnitine could be used for diagnosis and evaluating efficacy with such patients. Hyperbaric oxygen supplemented with qingkailing can relieve coma, correct metabolic disorders and accelerate patients′ awakening. Metabolomics provides a new method for identifying endogenous metabolic abnormalities in patients with post-traumatic consciousness disorders. It can be useful in prognosis and clinical treatment.

Objective:To construct a metrological safety evaluation model for large medical equipment based on the perspective of metrological safety,and to explore its application value in the management of large medical equipment.Methods:According to the different safety risk sources,the diagnosis and treatment area of large medical equipment was divided into five types:light source,electricity,water,radiation and information.The metrological safety assessment indicators were empowered by combining subjective and objective methods,and engineering support strategies were developed according to the metrological safety levels and safety change trends.A total of 48 large medical equipment in clinical use in The 960th Hospital of PLA Joint Logistic Support Force from 2020 to 2023 were selected.The 42 sets of equipment used during 2020 to 2021 adopted the conventional fault maintenance support mode(referred to as the conventional support mode),and the 43 sets of equipment used from 2022-2023(including 37 in conventional mode and 6 newly added)adopted the metrology safety technical support mode(referred to as metrology safety support mode).The metrology safety management ability score,metrological inspection qualification levels and metrological safety management effects of large medical equipment were compared between the two management modes.Results:The evaluation scores of theoretical knowledge,safety awareness,practical operation and emergency level indicators of large medical equipment use management personnel of the metrology safety support model were(17.39±1.38)points,(17.87±1.08)points,(27.68±1.45)points and(27.96±1.24)points,respectively,which were higher than those of the conventional support mode,the difference was statistically significant(t=4.234,4.371,5.832,3.764,P＜0.05).The average metrological inspection qualified rates(％)of medical imaging equipment,life support equipment,clinical laboratory equipment,operating room equipment and other equipment using the metrology safety support mode were(95.19±2.56)％,(95.56±1.79)％,(94.64±2.42)％,(96.20±2.01)％and(93.46±2.22)％,respectively,which were higher than those of the conventional support mode,the difference was statistically significant(t=3.846,2.809,2.405,2.492,2.742,P＜0.05).The proportion of large medical equipment rated as high safety level and low safety level in the metrological safety support model were 67.44％(29/43)and 4.65％(2/43),respectively,the proportion of high safety level was higher than that in the conventional support mode,and the proportion of low safety level was lower than that in the conventional support mode,the difference was statistically significant(x2=4.261,7.085,P＜0.05).Conclusion:The large medical equipment metrological safety support mode based on the safety perspective can improve the safety management ability of personnel related to equipment use and management,improve the operation quality and performance level of equipment,increase the proportion of high-safety level metrology safety of large medical equipment,and reduce the proportion of low-safety level.

Objective·To investigate the composition of immune cells in tumor microenvironment of colorectal cancer(CRC),and examine the proportion,phenotype and effector function of natural killer(NK)cells in CRC.Methods·Fresh tumor tissues,paired normal tissues adjacent to tumor,and peripheral blood samples in the same cohort were collected from CRC patients.Tissues were digested and prepared into single cell suspension.The major immune cell lineages were detected by flow cytometry.t-Distributed stochastic neighbor embedding(t-SNE)and statistical analysis were used to analyze the composition of immune cells in tumor microenvironment of CRC.To analyze the phenotype of NK cells,the expression levels of activation markers,including CD16,CD27,CD69,human leukocyte antigen-DR(HLA-DR),and T cell immunoglobulin domain and mucin domain-3(TIM-3),were detected by flow cytometry.NK cell subsets:CD38lowNK cells and CD38highNK cells were also examined by flow cytometry.To assess the effector function of NK cells,they were stimulated with cell stimulation cocktail and the expression levels of interferon-γ(IFN-y),tumor necrosis factor-α(TNF-α),and granulocyte-macrophage colony-stimulating factor(GM-CSF)were measured by flow cytometry.Results·Twenty-five pairs of fresh tumor tissues and normal tissues adjacent to tumor,and 15 peripheral blood samples in the same cohort were collected from CRC patients.The tumor microenvironment of CRC included diverse immune cell types,including T cells,B cells,NK cells and myeloid cells.The proportions of T cells(P=0.000)and myeloid cells(P=0.026)in tumor tissues were significantly higher than those in normal tissues.By contrast,the proportion of NK cells(P=0.007)in tumor was significantly reduced.The proportion of B cells was comparable between tumor and normal tissues.Compared to normal tissues,NK cells in tumor tissues expressed significantly lower CD27(P=0.000)and CD69(P=0.001),while the expression levels of CD16(P=0.008),HLA-DR(P=0.000)and TIM-3(P=0.024)were significantly elevated.The results indicated that NK cells in CRC tumor exhibited a phenotype of late activation and exhaustion.According to the expression level of CD38,NK cells could be divided into two subsets,CD38highNK cells and CD38lowNK cells.The proportion of CD38highNK cells(P=0.003)in tumor tissues was significantly lower than that in normal tissues,while the proportion of CD38lowNK cells was unaffected.Compared to CD38lowNK cells,CD38highNK cells expressed higher CD27,meanwhile significantly less CD16,NKp46,CD57,CD94,HLA-DR and CD158a(P=0.000).These results suggested that CD38highNK cells were at early differentiation state.The secretion of cytokines IFN-y(P=0.032),TNF-α(P=0.042),and GM-CSF(P=0.019)by tumor-infiltrated NK cells was significantly decreased compared to that in normal tissues.The results showed that the function of tumor-infiltrated NK cells was impaired.Conclusion·Together,these data suggest that NK cell compartment is disrupted in tumor tissues of CRC,leading to the impaired anti-tumor immunity mediated by NK cells.