Objective To investigate the application of organoid technology in colorectal cancer research and analyze the factors influencing the success rate of organoid cultivation. Methods A total of 24 samples of colorectal cancer patients treated at Tianjin Fifth Central Hospital and cultured organoids using specific culture media and Matrigel. The samples were collected within 30 minutes post-excision and stored at 4℃ to minimize contamination and protein degradation. During the cultivation process,the authors recorded instances of bacterial or fungal contamination and the organoid growth,and test for their histological structure and expression of tumor markers. Additionally,by analyzing clinical information from the patients who provided the samples,explore potential factors that may affect the success rate of culturing colorectal cancer organoids. Results A success rate of 70.8％ (17/24) was achieved in the cultivation of organoid. The success rate of organoid culture was significantly different from that of tumor stage (all P<0.05),with significantly higher successful organoid cultivation rate for stage Ⅱ and stage Ⅲ tumor tissues than those for stageⅠand Ⅳ[83.3％ (5/6),90.9％ (10/11) vs. 33.4％ (1/3),25.0％ (1/4)]. Additionally,samples with a Ki-67 positive area proportion of 55％-70％ exhibited the highest success rate (100％). Phenotypic experiments on the organoids indicated that their pathological histological structure and the expression of tumor markers were consistent with those of the primary tissues,suggesting that the organoids retained the histological characteristics of the primary lesions. Conclusions This study successfully established a colorectal cancer organoid model revealing the impact of tumor staging and the proportion of Ki-67 positive areas on the success rate of culture. The organoid model effectively retains the histological characteristics of the primary lesion,providing a powerful in vitro tool for the research and treatment of colorectal cancer.

Objective To investigate the application of organoid technology in colorectal cancer research and analyze the factors influencing the success rate of organoid cultivation. Methods A total of 24 samples of colorectal cancer patients treated at Tianjin Fifth Central Hospital and cultured organoids using specific culture media and Matrigel. The samples were collected within 30 minutes post-excision and stored at 4℃ to minimize contamination and protein degradation. During the cultivation process,the authors recorded instances of bacterial or fungal contamination and the organoid growth,and test for their histological structure and expression of tumor markers. Additionally,by analyzing clinical information from the patients who provided the samples,explore potential factors that may affect the success rate of culturing colorectal cancer organoids. Results A success rate of 70.8％ (17/24) was achieved in the cultivation of organoid. The success rate of organoid culture was significantly different from that of tumor stage (all P<0.05),with significantly higher successful organoid cultivation rate for stage Ⅱ and stage Ⅲ tumor tissues than those for stageⅠand Ⅳ[83.3％ (5/6),90.9％ (10/11) vs. 33.4％ (1/3),25.0％ (1/4)]. Additionally,samples with a Ki-67 positive area proportion of 55％-70％ exhibited the highest success rate (100％). Phenotypic experiments on the organoids indicated that their pathological histological structure and the expression of tumor markers were consistent with those of the primary tissues,suggesting that the organoids retained the histological characteristics of the primary lesions. Conclusions This study successfully established a colorectal cancer organoid model revealing the impact of tumor staging and the proportion of Ki-67 positive areas on the success rate of culture. The organoid model effectively retains the histological characteristics of the primary lesion,providing a powerful in vitro tool for the research and treatment of colorectal cancer.

Objective:To investigate the application value of three-dimensional (3D) recons-truction combined with endoscopic ultrasonography (EUS) in preoperative accurate evaluation of biliary tract neoplasms.Methods:The retrospective and descriptive study was conducted. The clinico-pathological data of 19 patients with biliary tract neoplasms who underwent 3D reconstruction combined with EUS in the Shangdong Provincial Third Hospital from January 2019 to October 2022 were collected. There were 13 males and 6 females, aged 64(range, 35-75)years. All patients underwent preoperative abdominal enhanced computer tomography (CT) thin-slice scan with 3D reconstruction combined with EUS. Some patients further received other endoscopic techniques such as intraductal ultrasonography, endoscopic retrograde cholangiopancreatography or SpyGlass cholangioscopy to obtain tumor tissues for histopathology evaluation. The surgical implementation protocol was developed based on the results of 3D reconstruction and EUS. Observation indicators: (1) results of 3D reconstruction; (2) results of EUS; (3) comparison between preoperative surgical protocol and actual intraoperative conditions. Measurement data with skewed distribution were represented as M(range), and count data were described as absolute numbers and/or percentages. Results:(1) Results of 3D reconstruction. Results of 3D reconstruction in 19 patients with biliary tract neoplasms showed morphology of the liver, bile ducts, pancreas, blood vessels, and duodenum, including 4 cases of hilar cholangiocarcinoma, 14 cases of middle and lower cholangiocarcinoma, and 1 case of intrahepatic cholangiocarcinoma. The accuracy of 3D reconstruction in 19 patients was 18/19. (2) Results of EUS. All 19 patients underwent preoperative EUS, including 7 cases obtained tumor tissue for histopathology evaluation, with the results indicating abnormal hyperplasia or malignant tumor. The rate of histopathology evaluation was 7/19, with the sensitivity as 7/7. Of 19 patients, results of EUS in 2 cases indicated positive of lymph node metastasis, but results of postoperative histopathology evaluation indicated negative of lymph node metastasis in lymph node specimens. Results of EUS in the rest of 17 cases indicated negative of lymph node metastasis, but results of intraoperative laparoscopic exploration on 1 case indicated extensive intra-abdominal metastasis. (3) Comparison between preoperative surgical protocol and actual intraoperative conditions. Of 19 patients, 18 cases underwent radical resection and 1 case underwent bile duct drainage, with the compliance rate between preoperative surgical protocol and actual intraoperative conditions as 18/19. The volume of intraoperative blood loss in the 18 cases receiving radical resection was 336(range, 50-1500)mL. Two cases had postoperative complications.Conclusion:Results of 3D reconstruction combined with EUS can accurately map the the size, location, extent of bile duct invasion, and adjacent relationships of surrounding tissues of malignant biliary tract neoplasms, for preoperative accurate evaluation and surgical planning.

Objective::To build a reference fetal growth chart for the Chinese population based on fetal ultrasound measurements.Methods::This was a multicenter, population-based retrospective cohort study. Longitudinal ultrasound measurement data were collected from 24 hospitals in 18 provinces of China from 1 st September through 31 st October of 2019. The estimated fetal weight (EFW) was calculated based on head circumference, abdominal circumference, and femur length using Hadlock formula 3. Fetal growth curves were estimated using a two-level linear regression model with cubic splines. All participants were divided into two groups: the northern group ( n = 5829) and the southern group ( n = 3246) based on the geographical division of China and male fetus group ( n = 4775) and female fetus group ( n = 4300) based on fetal gender. The EFW was compared by fetal gender and geographical group. All statistical models were adjusted for maternal sociodemographic characteristics. Results::A total of 9075 participants with 31,700 ultrasound measurement records were included in this study. Male fetuses demonstrated significantly larger EFW compared to female ones starting at 16 weeks of gestation and extending to delivery (global test P < 0.01). The overall geographic difference in EFW was significant (global test P = 0.03), and week-specific comparisons showed that the northern group had a greater EFW starting at 15 weeks of gestation and extending to 29 weeks of gestation, although this difference did not extend to the time of delivery. The Z-score of EFW confirmed that our Chinese fetal growth charts differed from previously published standards. Conclusion::This study provides EFW and ultrasound biometric reference measurements for Chinese fetuses and reveals differences from other fetal growth charts. The chart is worth promoting in more regions of China but should be tested prudently before use.

Objective::To build a reference fetal growth chart for the Chinese population based on fetal ultrasound measurements.Methods::This was a multicenter, population-based retrospective cohort study. Longitudinal ultrasound measurement data were collected from 24 hospitals in 18 provinces of China from 1 st September through 31 st October of 2019. The estimated fetal weight (EFW) was calculated based on head circumference, abdominal circumference, and femur length using Hadlock formula 3. Fetal growth curves were estimated using a two-level linear regression model with cubic splines. All participants were divided into two groups: the northern group ( n = 5829) and the southern group ( n = 3246) based on the geographical division of China and male fetus group ( n = 4775) and female fetus group ( n = 4300) based on fetal gender. The EFW was compared by fetal gender and geographical group. All statistical models were adjusted for maternal sociodemographic characteristics. Results::A total of 9075 participants with 31,700 ultrasound measurement records were included in this study. Male fetuses demonstrated significantly larger EFW compared to female ones starting at 16 weeks of gestation and extending to delivery (global test P < 0.01). The overall geographic difference in EFW was significant (global test P = 0.03), and week-specific comparisons showed that the northern group had a greater EFW starting at 15 weeks of gestation and extending to 29 weeks of gestation, although this difference did not extend to the time of delivery. The Z-score of EFW confirmed that our Chinese fetal growth charts differed from previously published standards. Conclusion::This study provides EFW and ultrasound biometric reference measurements for Chinese fetuses and reveals differences from other fetal growth charts. The chart is worth promoting in more regions of China but should be tested prudently before use.

Objective:To evaluate the safety and long-term effect of total parathyroidectomy in patients with secondary hyperparathyroidism.Methods:One hundred fifty-four patients with secondary hyperparathyroidism who underwent total parathyroidectomy in Zhongshan Hospital,Dalian University from Mar 2012 to Mar 2018 were followed up for 3-9 years,including the level of iPTH, serum calcium and phosphorus and dosing of calcium supplement.Results:Among the 154 patients, the iPTH level in 149 patients fluctuated within 15-60 pg/ml. After oral calcium carbonate, the blood calcium fluctuated in 1.8-2.4 mg/ml, and serum phosphorus was 0.8-1.6 mg/ml. The level of iPTH in 5 patients was between 80-150 pg/ml, which was higher than the normal value 10-70 pg/ml. The clinical symptoms of all patients were significantly relieved.Conclusion:Total parathyroidectomy is safe and reliable in the treatment of secondary hyperparathyroidism with low recurrence rate and stable long-term effect.

Objective To assess the effectiveness of repeated trans-cranial magnetic stimulation ( rTMS) in relieving post-stroke depression ( PSD). Methods PubMed, Embase, the Cochrane library, Web of Science, CNKI, WANFANG, and VIP were searched for reports of randomized, controlled trials of rTMS treatment of PSD published before June 2015. Crude standardized mean differences ( SMDs) and odds ratios with 95% confidence in-tervals ( CIs) were calculated for depression intensity and effectiveness rate after treatment using random or fixed effects models. Results Twenty-four studies involving 856 rTMS-treated patients and 802 control patients were in-cluded in the meta-analysis. The results showed that compared with the control group, PSD patients showed significant reductions in depression after rTMS treatment ( SMD=-1.36;95% CI-1.6 to-1.12;P≤0.05) . The total effective-ness rate in the treated group was 85% with a reduction in NIHSS score ( SMD=-0.82;95% CI-1.2 to-0.44;P≤0.05) . Subgroup analysis showed that neither the frequency of rTMS stimulation, the site stimulated, nor time after stroke had a significant influence on the effectiveness of rTMS. Additionally, a few studies reported adverse reactions after rTMS. Conclusion rTMS appears to be a safe and effective therapy for PSD. Further well-controlled trials may elucidate the mechanism underlying the placebo effects of the sham rTMS observed among PSD patients.

Objective:To valuate the treatment value and analyse the effect on the cellular immune functions by studying the differences of T-lymphocyte subsets and CD4+CD25+Treg cells in peripheral blood after adoptive immunotherapy ( dendritic cells and cytokine-induced killer cells,DC-CIK) combined with chemotherapy on MM.Methods:50 patients with MM were randomly divided into two groups.24 patients in chemotherapy group were treated by chemotherapy only,26 patients in joint group were treated by adoptive immunotherapy( DC-CIK) combined with chemotherapy,and the clinical outcomes and the levels of T-lymphocyte subsets and CD4+CD25+Treg cells in peripheral blood between two groups were compared.Moreover,the differences of cellular immune indicators (Th1/Th2,the ratio of AgNOR,and TGF-β)between two groups were also compared.Results: After treatment,quality of life,clinical index and survival in joint group were better than in chemotherapy group( P<0.05);the proportion of CD3+CD8+,the ratios of CD4+CD25+,CD4+CD25+/CD4+and the level of TGF-βof joint group wes clearly lower than chemotherapy group(P<0.05),and the ratios of CD3+CD4+/CD3+CD8+, Th1/Th2 and AgNOR of joint group wes clearly higher than chemotherapy group .Conclusion: DC-CIK combined with chemotherapy could be an effective and promising treatment to patients with MM,and it maybe strengthen the anti-tumor action of bodies by regulating the balance between Th1 and Th2 reaction.

BACKGROUND:Clinical infusion of hematopoietic stem cells and mesenchymal stem cells for treatment of aplastic anemia has been reported. OBJECTIVE:To investigate the effect of human adipose-derived mesenchymal stems cells on the secretion function of T lymphocytes of aplastic anemia patients. METHODS:Human adipose-derived mesenchymal stems cells were extracted from healthy human adipose tissues. T-lymphocytes were harvested from peripheral blood of patients with aplastic anemia by density gradient centrifugation. Human adipose-derived mesenchymal stems cells were co-cultured with T-lymphocytes. The levels of interleukin-2, interleukin-4, interleukin-10 and interferon-γwere detected by enzyme linked immunosorbent assay. T-bet and GATA-3 levels were examined by real-time PCR and western blot. RESULTS AND CONCLUSION:The levels of Th1 type cytokines interferon-γand interleukin-2 in the co-culture group were significantly lower than those in the T-lymphocyte group (P<0.05). But the levels of Th2 type cytokines interleukin-4 and interleukin-10 in the co-culture group were significantly higher than those in the T-lymphocyte group (P<0.05). The T-bet mRNA and protein levels in the co-culture group were significantly lower than those in the T-lymphocyte group, while the GATA-3 mRNA and protein levels were significantly higher in the co-culture group. Human adipose-derived mesenchymal stems cells can mediate an immunoregulation effect on T-lymphocytes of aplastic anemia patients in vitro, which is possibly related with the inhibition of Th1-dominant response due to the disorder of T-bet and GATA-3 gene expression.

Objective To explore the effects of mesenchymal stem cells ( MSC ) treatment on platelet counts in mice with immune-mediated thrombocytopenia ( ITP) and the possible mechanism .Meth-ods ITP was induced by daily intraperitoneal injection of anti-platelet membrane CD 41 antibody (MWReg30) into BALB/c mice.The mice were then divided into experiment and control groups with 20 mice in each.Each mouse in experimental group was injected with 2×107 mesenchymal stem cells (MSC) through the tail vein .The numbers of blood platelets in mice from two groups were counted on days 5, 7 and 14 after MSC injection .Reverse transcriptase polymerase chain reaction ( RT-PCR) was performed to meas-ure T-bet and GATA-3 gene expression in peripheral blood mononuclear cells ( PBMCs ) at mRNA level on day 14.The levels of IFN-γ, IL-2, IL-4 and IL-10 in serum were detected by ELISA .Results The platelet counts in experimental group were significantly higher than those in control group on days 7 and 14 after MSC injection [(588.0±81.6)×109/L and (623.0±78.9) ×109/L vs.(317.0±90.1) ×109/L and (288.0± 87.8)×109/L ] (P<0.05).On day 14 after MSC injection, the T-bet expression at mRNA level in PBMCs from mice in experimental group was significantly lower than that in control group [(0.04±0.03) vs.(0.27 ±0.05)] (P<0.05), while the GATA-3 expression at mRNA level was higher than those in control group [ (0.14±0.04) vs.(0.07±0.05)] (P<0.05).Compared with control group, the concentrations of Th1 type cytokines such as IFN-γand IL-2 were remarkably down-regulated in experimental group [(3.1±1.7) pg/ml and (3.2±2.1) pg/ml vs.(10.3±4.8) pg/ml and (16.3±5.7) pg/ml](P<0.05), while the con-centrations of Th2 type cytokines such as IL-4 and IL-10 were up-regulated in experimental group [(88.6± 15.2) pg/ml and (38.3±11.8) pg/ml vs.(32.7±5.7) pg/ml and (22.1±3.4) pg/ml ] (P<0.05). Conclusion MSC treatment can effectively increase platelet counts in mice with immune-mediated thrombo-cytopenia, which may be associated with the suppression of Th 1-dominant response mediated by abnormal ex-pression of T-bet and GATA-3.