Objective To construct an in vitro co-culture model of gastric cancer organoids and cancer-associated fibroblasts(CAFs),and investigate the role of cancer-associated fibroblasts in the proliferation and chemotherapy resistance of gastric cancer organoids.Methods Tumor tissues from 12 gastric cancer patients undergoing surgical treatment in Department of General Surgery of Second Affiliated Hospital of Army Medical University from February 2023 to March 2024 were collected to construct gastric cancer organoids using 3D culture.HE staining was used to observe the morphology,and immunohistochemical assay was employed to determine the expression of cytokeratin CK7,carcinoembryonic antigen(CEA),and proliferation marker Ki-67.After CAFs derived from the same patient were cultured,observed for their morphology under a light microscope,and detected for the phenotype by flow cytometry,the cells were co-cultured with gastric cancer organoids in a 1:1 ratio.Phase-contrast microscopy was applied to observe the growth of the organoids and analyze the number,average diameter,and total area.Then,organoids cultured alone served as the control group.After the control and co-culture groups were treated with chemotherapy drugs,5-fluorouracil and oxaliplatin,for 48 h,the viability and apoptosis of organoids were assessed with CellTiter-Glo??3D assay and CellEvent? Caspase 3/7 activity,respectively.Results Gastric cancer organoids and CAFs were successfully established from 10 gastric cancer patient-derived samples.The gastric cancer organoids exhibited morphological characteristics consistent with the corresponding primary tumors,and showed positive expression of CK7,CEA,and Ki-67.CAFs displayed typical spindle-shaped morphology and exhibited the phenotypic markers CD326-,CD45-,CD31-,α-SMA+,CD73+,CD90+,and CD105+.Compared to the organoids cultured alone,the organoids co-cultured with CAFs showed more formation of organoids,in larger average diameter,and taking larger total area(P<0.05).After the treatment of 5-fluorouracil and oxaliplatin,the half-maximal inhibitory concentration(IC50)was 10.66 and 3.26 μmol/L,respectively in the control group,while was 46.23 and 91.11 μmol/L in the co-culture group.Additionally,the number of CellEvent? Caspase 3/7 positive apoptotic cells was significantly less in the co-culture group than the control group.Conclusion Compared with individually cultured gastric cancer organoids,the co-culture model of gastric cancer organoids and CAFs better simulates the pro-tumor proliferation and drug resistance effects of in vivo tumor microenvironment.

Middle-aged and elderly patients with femoral neck fracture often suffer from basic diseases. Conservative treatment will significantly increase the incidence of complications. At present, surgical treatment is mostly advocated. Internal fixation is one of the effective treatment methods for middle-aged and elderly patients with femoral neck fracture. It has the advantages of improving hip joint function, accelerating patient recovery, and improving patient quality of life. At present, there are many choices of internal fixation in the world, each has its own advantages and disadvantages, and there is no unified standard. Different surgical methods can be selected according to various factors such as fracture type, patient′s physical condition and surgical auxiliary technology. This article reviews the clinical progress of internal fixation for femoral neck fractures in middle-aged and elderly patients from four aspects: the characteristics of femoral neck fractures in middle-aged and elderly patients, common internal fixation methods, 3D printing guide plate auxiliary technology and artificial intelligence auxiliary technology.

Objective: To investigate the mechanism of action of Wuzi Yanzong pill (WYP) in rats with oligoasthenozoospermia (OAZ) via metabolomics and to provide a possible basis for improving this WYP-based treatment. Methods: A rat model of OAZ was established by treating male Sprague–Dawley rats with glucosides from Tripterygium wilfordii Hook. F. Seventy-two rats were randomly divided into six groups: control, L-carnitine (positive control), model, and low-, medium-, and high-dose WYP groups. Rats in the experimental groups were treated with WYP for 4 weeks. At the end of the treatment period, sperm cell quality (density, motility, and viability) was assessed using a semen analysis system, mitochondrial membrane potential (MMP) was assessed using flow cytometry, and testicular injury was assessed using hematoxylin and eosin staining to validate the therapeutic effect of WYP in OAZ. Further, serum metabolomics-based analysis was performed using high-performance liquid chromatography-mass spectrometry to identify differential metabolic pathways and possible mechanisms of action of WYP in OAZ treatment. Results: A rat model of OAZ was considered successfully-established after comparing the quality of spermatozoa in the model group to that in the control group. WYP-M and WYP-H treatments significantly improved sperm cell density, motility, and viability compared with those in the model group (all P < .05). Compared with the model group, both WYP-M and WYP-H treatments increased MMP values (P = .006 and P = .021 respectively), while there was no significant difference in the L-carnitine group. L-carnitine and WYP administration reversed damage to the testes to varying degrees compared with that in the model group. Further, 44 differential metabolites and four metabolic pathways, especially autophagy pathway, related to OAZ were identified via metabolomics. Conclusions WYP improves sperm cell quality and MMP in OAZ primarily via autophagy regulation. These findings can be employed to improve the efficacy of WYP in humans.

Objective To investigate the distribution of B cells in both tumor and non-tumor tissues of gastric cancer patients,analyze their phenotypic characteristics and explore the impact on T cell proliferation.Methods Immunohistochemical staining was utilized to detect the expression of B cell surface marker CD 19 in tumor and non-tumor tissues from 33 gastric cancer patients.The expression levels of chemokine receptors and immunoglobulin molecules on B cells in both tumor and non-tumor tissues were measured using flow cytometry.Chemotaxis experiments were conducted to examine the role of the CXCL12-CXCR4 axis in B cell chemotaxis.B cells isolated and purified from both tissue types were co-cultured with autologous peripheral T cells to assess their effect on T cell proliferation.Results There were significantly more B cells infiltrated in tumor tissues than those infitrated in the non-tumor tissues of gastric cancer patients(P＜0.01),and CXCR4 was highly expressed on tumor-infiltrating B cells compared with B cells derived from non-tumor tissues(P＜0.05).The Cancer Genome Atlas(TCGA)analysis indicated that the expression level of CXCL12 in tumor tissues was positively correlated with the expression level of CD19 in gastric cancer patients(r=0.15,P＜0.01).And the expression level of CXCL12 in tumor tissues of the gastric cancer patients was also positively correlated with the number of B cells infiltrated in tumor tissues.Chemotaxis experiments confirmed that the CXCL12-CXCR4 axis was involved in promoting B cell chemotaxis(P＜0.05).Although B cells in tumor and non-tumor tissues had similar levels of IgM,IgG,and IgA expression,tumor-infiltrating B cells significantly inhibited the proliferation of T cells when compared with B cells derived from non-tumor tissues(P＜0.01).Conclusion There are more B cells infiltrated in gastric cancer tissues,which may be recruited to tumor tissues through the CXCL12-CXCR4 axis,and then inhibit T cell proliferation to promote the progression of gastric cancer.

Objective:To evaluate the shoulder arthroscopic treatment of avulsion fracture of humeral greater tuberosity with double-row suture bridge.Methods:Retrospectively analyzed were the 13 avulsion fractures of humeral greater tuberosity which had been treated by shoulder arthroscopic double-row suture bridge at Bone and Joint Department, Guangzhou Liwan District Orthopedic Hospital from March, 2018 to March, 2020. There were 6 males and 7 females, with an average ages of 52.3 years (from 35 to 69 years). According to the Mutch classification, all the fractures of humeral greater tuberosity were attributed to the avulsion type. Of them, 3 were simple avulsion fractures of humeral greater tuberosity and 10 were complicated with shoulder anterior dislocation which was reduced manually before operation. There were 3 obsolete avulsion fractures of humeral greater tuberosity. Regular clinic and X-ray follow-ups were carried out. At the final follow-up, the pain, range of motion (ROM) and stability of the shoulder joint were assessed using visual analogue scale (VAS), American shoulder elbow scores (ASES), and Korean shoulder scores (KSS).Results:All the patients were followed up for an average of 12.3 months (from 10 to 16 months) postoperatively. No infection or shoulder instability was observed. At the final follow-up, the abduction and elevation averaged 164.6° (from 135° to 180°), the lateral external rotation 62.7° (from 40° to 80°), the internal rotation touch back test T10 level (from L2 to T6), the VAS 0.65 (from 0 to 2.5), the ASES 90.5 (from 78 to 100), and the KSS 91.5 (from 84 to 100).Conclusion:Shoulder arthroscopic treatment of avulsion fracture of humeral greater tuberosity with double-row suture bridge is minimally invasive and allows for repair of combined injury, leading to quick postoperative recovery.

Objective:To observe the changes of macular morphology and best corrected visual acuity (BCVA) in children with retinopathy of prematurity (ROP) after single treatment of laser photocoagulation or intravitreal injection of ranibizumab (IVR) by optical coherence tomography angiography (OCTA).Methods:A cross sectional study was conducted.Fourteen cases (23 eyes) of ROP children with a treatment history aged 4-10 years were enrolled as the ROP group in Shenzhen Eye Hospital from April to August 2018, with 8 cases (16 eyes) of age-matched full-term children as control group.Among the ROP group, 9 cases (14 eyes) receiving photocoagulation were set as the photocoagulation group and 5 cases (9 eyes) receiving IVR as the injection group.The 3 mm×3 mm scanning mode of OCTA was adopted to measure the area of foveal avascular zone (FAZ), FAZ morphological index, superficial retinal vascular density (VD) and superficial retinal perfusion density (PD), and the B-scan mode was used to determine the central foveal thickness (CFT). BCVA of all children was recorded and analyzed.This study adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Shenzhen Eye Hospital (No.b-03). Written informed consent was obtained from guardians prior to any examination.Results:Compared with the control group, there was smaller area of FAZ, more irregular FAZ morphology and thicker CFT in the ROP group.The central fovea of macula in the laser photocoagulation group was shallow and almost indiscernible, and FAZ morphology was irregular.The FAZ area, FAZ morphological index, VD and PD in the ROP group were (0.09±0.08) mm 2, 0.51±0.25, (19.18±2.50)mm/mm 2 and 0.35±0.04, respectively, which were significantly lower than (0.32±0.08)mm 2, 0.68±0.07, (20.94±0.93)mm/mm 2, 0.38±0.02, respectively in the control group ( t=-8.03, -3.14, -3.02, -2.52; all at P<0.05). The CFT in the ROP group was (215.00±21.32) μm, which was significantly thicker than (164.29±20.20) μm in the control group ( t=8.91, P<0.05). There was no statistical difference in BCVA between the two groups ( t=0.16, P=0.87). The CET in the laser photocoagulation group was significantly thicker than that in the injection group ( t=3.50, P=0.01). Conclusions:There are obvious morphology changes of macula in ROP children with single treatment history of laser photocoagulation or IVR, while the BCVA is not significantly affected by the treatment.

The exact pathophysiological mechanisms of retinopathy of prematurity (ROP) remain elusive. The risk factors of ROP include excessive oxygen therapy, malnutrition, infection and inflammation. Among the factors above, the effect of inflammation on ROP has received more attention. TNF-α is a biological active protein which is involved in neovascularization and inflammation. It may play a role in the development of ROP. This review summarized the studies on the association between TNF-α and ROP in recent years, so as to provide a new way to further study the pathogenesis and treatment methods of ROP.

Objective To investigate the status of self‐management of blood pressure (BP) among elderly hypertensive patients living in new rural communities of Shanghai. Methods We enrolled a total of 2 241 hypertensive patients over 60 years old who participated in the National Key Research and Development Project "Cohort study on hypertension" of the 13th Five‐Year Plan in Langxia, Shanghai from May to July 2018. Questionnaires, physical examinations, and laboratory tests were conducted. We collected participants' demographic information, cardiovascular history, and self‐management behaviors related to hypertension. The status of self‐management of BP among participants with hypertension was analyzed. Pearson's chi‐squared test was used to explore the differences between males and females. Results Participants'mean age was (70.0±6.8) years, and males accounted for 38.8% of all participants. The systolic and diastolic BP levels of participants were (147.8±15.4) mmHg (1 mmHg=0.133 kPa) and (82.8±9.8) mmHg, respectively. The systolic blood pressure level of women was higher than that of men, and the diastolic blood pressure level of men was higher; the difference was statistically significant (all P<0.05). The awareness, treatment, and control rate of hypertension were 79.0%, 78.0%, and 25.6%, respectively. Males had a significantly higher control rate than females (P<0.05). The status of self‐management of blood pressure was as follows: (1) As to lifestyle, 84.4% of participants lacked exercise, and 69.2% were overweight or obese. The prevalence of smoking and drinking was higher in males (40.7% and 46.2%, respectively), and the proportion of females (12.4%) with anxiety and/or depression was higher than that of males (P all<0.001). (2) Concerning BP‐monitoring, only 10.1% of participants measured BP every week, 22.9% had BP measurement instruments at home, and 4.6% recorded their BP levels. There was no significant difference in self‐blood pressure monitoring behavior between men and women. (3) Regarding adherence to medication treatment, 97.6% took antihypertensive drugs regularly, and 21.6% were on combination therapy. The proportion of men taking combined drugs was higher than that of women, but there was no significant difference. Conclusions The rates of awareness, treatment, and control of hypertension in elderly hypertensive patients in new rural communities of Shanghai were above the national average. However, the status of self‐management of BP was still not satisfactory. More attention should be paid to healthy lifestyle, home BP monitoring, and effective pharmaceutical treatment strategies for hypertension.

Objective To observe the effect of retinal hemorrhage on the treatment of retinopathy of prematurity (ROP) by laser photocoagulation.Methods Retrospective case analysis.Screening and diagnosis of 134 eyes in 67 patients with ROP in Zone IⅡ Stage 3+ were included in the study.Among them,32 patients were male and 35 patients were female.The average birth gestational age was 27.80 ± 2.55 weeks.The average birth weight was 1060± 320 g.All children underwent binocular indirect ophthalmoscopy and RetCam Ⅲ.Of the 134 eyes,38 eyes (28％) with anterior,ridge or vitreous hemorrhage (group A);96 eyes (72％) without hemorrhage.Retinal avascular photocoagulation was performed within 72 hours after diagnosis by intravenous sedative combined with ocular surface anesthesia with 810 nm laser.Follow-up was performed at 1,4,8 and 12 weeks after treatment,and then every 6 months thereafter.The same equipment and methods before treatment were used to examine and document the regression and progression of ROP.The number of eyes with lesions after photocoagulation in the two groups was compared by x2 test.The t-test was used to compare the gestational age and birth weight.Results Among 134 eyes,lesions completely resolved in 125 eyes (93.3％),progressed in 9 eyes (6.7％).In group A,7 eyes were progressive (18.4％).In group B,2 eyes were progressive (2.1％).There was a statistically significant difference in the number of eyes with lesions after laser treatment in group A and B (x2=9.14,P=0.003).There was no significant difference in birth gestational age and birth weight (t=0.85,0.25;P=0.40,0.80).Conclusion The laser photocoagulation is safe and effective in the treatment of ROP.The preretinal,ridge or vitreous hemorrhage is related to the progression of the lesion after laser photocoagulation.

Objective To study the role and mechanism of orexin A in cell viability of Alzheimer's disease (AD) cell model PC12. Methods PC12 cells were treated with 0, 10, 20, 30, 40 and 50 μmol/L Aβ25-35 for 24 h, and then, cell viability was measured by MTT to confirm which concentration was the suitable one to establish the AD cell models. (1) AD cell models were treated with 0, 0.01, 0.1, 1 and 2μmol/L orexin A for 24 h, and then, 30μmol/L Aβ25-35 was added for 24 h; MTT assay was used to determine the cell viability to conform the suitable concentration of orexin A. (2) Inverted phase contrast microscope was employed to observe the morphology changes of PC12 cells from the control group, 30 μmol/L Aβ25-35 treatment group, and 0.01 μmol/L orexin A+30 μmol/L Aβ25-35 treatment group. (3) The PC12 cells were given pretreatment of orexin A receptor inhibitor SB408124 for 2 h, and cell viability was detected. Results (1) Aβ25-35 at concentration 30μmol/L was the suitable one to establish the AD cell models;after being pretreated with different concentrations of orexin A, the cell viability showed significant differences (F=27.120, P=0.000), and 0.01μmol/L orexin A was the suitable concentration. (2) Some of the cells from the 30μmol/L Aβ25-35 treatment group had breaking-off of protuberance and damaged soma;cells from 0.01μmol/L orexin A+30μmol/L Aβ25-35 treatment group had breaking-off of protuberance, and the degree of damaged soma was eased as compared with that in the 30μmol/L Aβ25-35 treatment group. (3) If the cell viability of the control group was 100％, cell viability of orexin A receptor inhibitor group was 109.10％±0.36％, which was significantly decreased as compared with that in the 0.01 μmol/L orexin A pretreated group (117.24％±2.72％, P<0.05). Conclusion Orexin A can improve the cell viability via combination of its specific receptor; orexin A and its specific receptor may be new targets for prevention and cure of AD.