Objective:To investigate the impact of postoperative complications on adverse outcomes following curative-intent resection for gallbladder cancer (GBC).Methods:The multi-center real-world study was conducted. The clinicopathological data of 629 patients with GBC, who were admitted to 14 medical centers including The First Affiliated Hospital of Army Medical University from the national multicenter database of Biliary Surgery Group of Elite Group of Chinese Journal of Digestive Surgery, from April 2020 to April 2024 were collected. There were 225 males and 404 females, aged (64±10)years. Patients underwent open curative-intent resection for GBC. Observation indicators: (1)surgery, postoperative complica-tions and adverse outcomes; (2) analysis of risk factors affecting postoperative adverse outcomes in patients and population attributable fraction (PAF). Missing data in predictor variables were addressed using multiple imputation with chained equations, while cases with missing outcome variables were addressed using the "multiple imputation then deletion (MID)" strategy. The severity of multicollinearity among independent variables was assessed using the variance inflation factor (VIF) test. Multivariable possion regression models with log link and robust error variance were construc-ted incorporating restricted cubic splines (3 knots) to address nonlinear relationships in continuous variables, calculating adjusted relative risk ( RR) with corresponding 95% confidence interval ( CI). Adjusted PAF was calculated for each imputed dataset using the AF package of R software, with subsequent pooling performed according to Rubin's rules. Results:(1) Surgery, postoperative complications and adverse outcomes. All 629 patients underwent curative-intent resection for GBC, of which 143 cases had postoperative complications, including 68 cases of intra-abdominal ascites, 39 cases of pulmonary infection, 21 cases of bile leakage, 12 cases of intra-abdominal hemorrhage, 11 cases of liver failure, 10 cases of pan-creatic fistula, 10 cases of wound infection, 10 cases of gastroparesis, 7 cases of cholangitis, 7 cases of sepsis. The same patient could have more than one kind of complication. Of 629 patients, there were 19 cases of postoperative 90-day death and 11 cases of missing data, 42 cases with post-operative 90-day reoperation and 7 cases with missing data, 44 cases with postoperative 90-day readmission and 3 cases with missing data, 155 cases with prolonged postoperative hospital stay and 3 cases with missing data. (2) Analysis of risk factors affecting the postoperative adverse outcomes in patients and PAF. Results of multivariate analysis showed that pulmonary infection and liver failure were independent risk factors for postoperative 90-day mortality ( RR=3.74, 12.15, 95% CI as 1.18-11.83, 1.98-74.48, P<0.05). Pulmonary infection demons-trated the highest PAF as 4.61% (95% CI as 3.94%-5.28%, P<0.05). Intra-abdominal ascites, pulmonary infection, bile leakage, and intra-abdominal hemorrhage were independent risk factors for post-operative 90-day reoperation ( RR=4.80, 3.62, 3.46, 4.99, 95% CI as 2.49-9.26, 1.42-9.21, 1.34-8.92, 1.55-16.06, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 8.65% (95% CI as 8.22%-9.08%, P<0.05). Intra-abdominal ascites, bile leakage, and liver failure were independent risk factors for postoperative 90-day readmission ( RR=6.20, 3.33, 14.33, 95% CI as 3.21-11.95, 1.33-8.35, 3.72-55.28, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 9.11% (95% CI as 8.85%-9.37%, P<0.05). Intra-abdominal ascites, pulmonary infection, bile leakage, liver failure, and wound infection were independent risk factors for prolonged postoperative hospital stay ( RR=2.29, 2.21, 2.26, 2.14, 3.35, 95% CI as 1.63-3.23, 1.41-3.46, 1.32-3.86, 1.11-4.13, 1.70-6.60, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 6.03% (95% CI as 5.71%-6.35%, P<0.05). Conclusion:Pulmonary infection is the most significant risk factor for postoperative 90-day mortality after curative-intent resection for GBC, while intra-abdominal ascites is the most significant risk factor for postoperative 90-day reoperation, postoperative 90-day readmission, and prolonged postoperative hospital stay.

Objective:To investigate the impact of postoperative complications on adverse outcomes following curative-intent resection for gallbladder cancer (GBC).Methods:The multi-center real-world study was conducted. The clinicopathological data of 629 patients with GBC, who were admitted to 14 medical centers including The First Affiliated Hospital of Army Medical University from the national multicenter database of Biliary Surgery Group of Elite Group of Chinese Journal of Digestive Surgery, from April 2020 to April 2024 were collected. There were 225 males and 404 females, aged (64±10)years. Patients underwent open curative-intent resection for GBC. Observation indicators: (1)surgery, postoperative complica-tions and adverse outcomes; (2) analysis of risk factors affecting postoperative adverse outcomes in patients and population attributable fraction (PAF). Missing data in predictor variables were addressed using multiple imputation with chained equations, while cases with missing outcome variables were addressed using the "multiple imputation then deletion (MID)" strategy. The severity of multicollinearity among independent variables was assessed using the variance inflation factor (VIF) test. Multivariable possion regression models with log link and robust error variance were construc-ted incorporating restricted cubic splines (3 knots) to address nonlinear relationships in continuous variables, calculating adjusted relative risk ( RR) with corresponding 95% confidence interval ( CI). Adjusted PAF was calculated for each imputed dataset using the AF package of R software, with subsequent pooling performed according to Rubin's rules. Results:(1) Surgery, postoperative complications and adverse outcomes. All 629 patients underwent curative-intent resection for GBC, of which 143 cases had postoperative complications, including 68 cases of intra-abdominal ascites, 39 cases of pulmonary infection, 21 cases of bile leakage, 12 cases of intra-abdominal hemorrhage, 11 cases of liver failure, 10 cases of pan-creatic fistula, 10 cases of wound infection, 10 cases of gastroparesis, 7 cases of cholangitis, 7 cases of sepsis. The same patient could have more than one kind of complication. Of 629 patients, there were 19 cases of postoperative 90-day death and 11 cases of missing data, 42 cases with post-operative 90-day reoperation and 7 cases with missing data, 44 cases with postoperative 90-day readmission and 3 cases with missing data, 155 cases with prolonged postoperative hospital stay and 3 cases with missing data. (2) Analysis of risk factors affecting the postoperative adverse outcomes in patients and PAF. Results of multivariate analysis showed that pulmonary infection and liver failure were independent risk factors for postoperative 90-day mortality ( RR=3.74, 12.15, 95% CI as 1.18-11.83, 1.98-74.48, P<0.05). Pulmonary infection demons-trated the highest PAF as 4.61% (95% CI as 3.94%-5.28%, P<0.05). Intra-abdominal ascites, pulmonary infection, bile leakage, and intra-abdominal hemorrhage were independent risk factors for post-operative 90-day reoperation ( RR=4.80, 3.62, 3.46, 4.99, 95% CI as 2.49-9.26, 1.42-9.21, 1.34-8.92, 1.55-16.06, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 8.65% (95% CI as 8.22%-9.08%, P<0.05). Intra-abdominal ascites, bile leakage, and liver failure were independent risk factors for postoperative 90-day readmission ( RR=6.20, 3.33, 14.33, 95% CI as 3.21-11.95, 1.33-8.35, 3.72-55.28, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 9.11% (95% CI as 8.85%-9.37%, P<0.05). Intra-abdominal ascites, pulmonary infection, bile leakage, liver failure, and wound infection were independent risk factors for prolonged postoperative hospital stay ( RR=2.29, 2.21, 2.26, 2.14, 3.35, 95% CI as 1.63-3.23, 1.41-3.46, 1.32-3.86, 1.11-4.13, 1.70-6.60, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 6.03% (95% CI as 5.71%-6.35%, P<0.05). Conclusion:Pulmonary infection is the most significant risk factor for postoperative 90-day mortality after curative-intent resection for GBC, while intra-abdominal ascites is the most significant risk factor for postoperative 90-day reoperation, postoperative 90-day readmission, and prolonged postoperative hospital stay.

AIM:To investigate the oncolytic effect of human umbilical cord mesenchymal stem cells(hUMSCs)delivering reovirus type 3(Reo3)on chronic myeloid leukemia(CML)K562 cells.METHODS:The expression of junc-tional adhesion molecule-A(JAM-A),a receptor susceptible to Reo3,on the surface of hUMSCs and K562 cells was as-sessed by flow cytometry.Intracellular viral inclusion body distribution 72 h after Reo3 infection in hUMSCs was observed by electron microscopy.The hUMSCs were infected with various multiplicities of infection(MOI)of Reo3(MOI=0,1,2 and 3)for 24,48,72,96 and 120 h,and the most suitable MOI was identified by CCK-8 assay.Subsequently,hUMSCs were infected with the optimal titer of Reo3 for the same durations,and supernatants were collected.The titer of Reo3 in the supernatant from each group was measured using mouse fibroblast L929 cells combined with median tissue culture in-fectious dose(TCID50)method,determining the optimal infection time.The K562 cells were divided into 4 groups:con-trol group,hUMSCs group,Reo3 group,and hUMSCs-Reo3 group.Ratios of hUMSCs to K562 cells in hUMSCs group and hUMSCs-Reo3 group were set at low,medium and high(5∶1,10∶1 and 20∶1).The changes of K562 cell viability af-ter co-cultured with hUMSCs-Reo3 for 24,48 and 72 h were analyzed by CCK-8 assay.The apoptosis of K562 cells was evaluated by flow cytometry.The half maximal effective concentration(EC50)of anti-Reo3 monoclonal antibody was deter-mined using L929 cells.The oncolytic effect of hUMSCs-Reo3 on K562 cells with antibody present in vitro was verified.Western blot analysis was used to detect the protein levels of Bcl-2,Bax,survivin and cleaved caspase-3 in K562 cells af-ter treatment.A BALB/c nude mouse subcutaneous tumor model was constructed with K562 cells(n=6)to analyze the in vivo anti-tumor effect of hUMSCs-Reo3.RESULTS:The expression levels of JAM-A on the surfaces of hUMSCs and K562 cells were found to be 11.0%and 99.0%,respectively.Electron microscopy revealed a significant presence of viral inclusion bodies within hUMSCs 72 h following infection with Reo3.Within 120 h,no statistically significant difference was observed in the viability of hUMSCs between Reo3(MOI=1)group and uninfected group,establishing the optimal MOI.The TCID50 results indicated that the highest virus titer in the lysate of hUMSCs in Reo3(MOI=1)group occurred 48 h after infection,determining 48 h as the optimal infection time.The K562 cells co-cultured with hUMSCs-Reo3 for 24,48 and 72 h showed a dose-and time-dependent inhibition of cell viability.The EC50 of the anti-Reo3 monoclonal antibody was found to be 1∶34.Even in the presence of antibodies at various concentrations(1∶34,1∶300 and 1∶600),hUMSCs were capable of transporting Reo3 to inhibit K562 cell viability and induce apoptosis in vitro.Compared with control group,significant down-regulation of Bcl-2 and survivin expression levels in K562 cells was noted after 48 h of co-culture with hUMSCs-Reo3(P<0.05),while Bax and cleaved caspase-3 expression levels were significantly up-regulated(P<0.05 or P<0.01).In the BALB/c nude mouse tumor-bearing model,determination of tumor volume changes,pathological examination of tumor tissue and major organs,and assessment of cathepsin B/L activity using a small animal live imaging system confirmed the oncolytic effect of hUMSCs-Reo3 on K562 cells in vivo without adverse effects on normal tissues.CONCLUSION:The hUMSCs are effective in transporting Reo3,and this delivery system is capable of releasing suffi-cient quantities of Reo3 in both in vivo and in vitro settings to inhibit the malignant proliferation of K562 cells and promote apoptosis,thereby exerting an oncolytic effect.

AIM:To explore the synergistic sensitization effect of human umbilical cord mesenchymal stem cell culture supernatant(hUMSC-CM)combined with temozolomide(TMZ)on various glioma cell lines,and to elucidate the underlying mechanisms.METHODS:The hUMSC-CM was harvested using two different serum deprivation tech-niques at 24 and 48 h,and was converted into freeze-dried powder,which was then given to rat malignant glioma cell line RG-2,human astrocytoma cell line U251 and human glioblastoma cell line LN-428 at 5 concentrations(0,1,3,6 and 9 g/L).The effectiveness and sensitivity of hUMSC-CM for inhibiting growth of glioma cells at 24,48 and 72 h were as-sessed using CCK-8 assay.Hematoxylin-eosin(HE)staining combined with CCK-8 assay was employed to evaluate the chemotherapy sensitivity of glioma cells after 48 h of treatment with TMZ at 6 concentrations(0,25,50,100,200 and 400 μmol/L).Two concentrations(3 and 9 g/L)of hUMSC-CM and 3 concentrations(50,100 and 200 μmol/L)of TMZ were chosen for concurrent treatment of glioma cells to assess the proliferation and pathological alterations.TUNEL staining was utilized to detect apoptosis.Flow cytometry was utilized to analyze cell cycle modifications.The expression alterations of apoptosis-inducing proteins,cleaved caspase-3,cleaved caspase-8 and cleaved PARP1,as well as autophagy-inducing proteins beclin-1 and LC3,were examined using Western blot to investigate the synergistic sensitization mechanism of hUMSC-CM combined with TMZ in vitro.RESULTS:The susceptibility of glioma cell lines to hUMSC-CM and TMZ varied,with RG-2 showing the highest sensitivity,followed by U251,and then LN-428.The inhibitory effect of hUMSC-CM(3 and 9 g/L)and TMZ(50,100 and 200 μmol/L)combined treatment on glioma cells was significantly greater than that that of single-agent treatments(P＜0.05),demonstrating a dose-and concentration-dependent enhancement.Notably,the combination of 9 g/L hUMSC-CM(C9)with 50 μmol/L TMZ(T50)effectively suppressed glioma cell growth.CCK-8 as-say indicated a significant reduction of cell viability in C9+T50 group compared with either C9 or T50 alone(P＜0.05).HE staining and TUNEL staining revealed pronounced morphological changes and significant apoptotic features in glioma cells treated with C9+T50.Flow cytometric analysis confirmed that C9+T50 induced cell cycle arrest in glioma cells.Fur-thermore,compared with control group,the levels of cleaved caspase-3,cleaved caspase-8,cleaved PARP1,beclin-1,and LC3-Ⅱ/LC3-Ⅰ were significantly elevated in the C9+T50-treated glioma cells(P＜0.01).CONCLUSION:(1)The concomitant administration of hUMSC-CM and TMZ exerts a broad inhibitory effect on glioma cells,with a synergistic sen-sitization observed across different cell lines.(2)The enhancement of glioma cell sensitivity to TMZ by hUMSC-CM may be attributed to the modulation of caspase-8/caspase-3/PARP1 signaling pathway and the induction of both apoptosis and autophagy in glioma cells.

OBJECTIVE: To explore the pathological characteristics and significance of RET proto-oncogene screening in multiple endocrine neoplasia type 2A (MEN2A) with cutaneous lichen amyloidosis (CLA). METHODS: Clinical data of 51 members from 7 unrelated pedigrees of MEN2A-CLA were collected. Systemic clinical investigations including biochemical testing, imaging examination, germline RET variant screening and histopathological examination were carried out. RESULTS: RET gene variants were detected in 28 patients with MEN2A (C634G/F/R/S/W and C611Y) including 12 males and 16 females, with the mean age of diagnosis being (41.1 ± 18.3) years old, which were consistent with their clinical manifestations. The incidence of medullary thyroid carcinoma (MTC), pheochromocytoma (PHEO), hyperparathyroidism (HPTH) and CLA among 28 MEN2A patients were 89.3%, 28.6%, 7.1% and 28.6%, respectively. Comparison of the incidence of MTC/PHEO/HPTH and CLA between C611Y and C634G/F/R/S/W, only PHEO and CLA in C611Y were lower than those in C634G/F/R/S/W (P < 0.05; P < 0.05). Among 8 patients with CLA, the male to female ratio was 2 : 6. The clinical features included pruritus in the interscapular region and presence of dry, thickened, scaly, brown pigment, clustered or desquamate-like plaques. The mean onset age of CLA [(18.4 ± 4.6) years] versus the mean age at diagnosis of CLA or MEN2A were significantly different (P < 0.001; P < 0.001). CONCLUSION MEN2A-CLA may be the early clinical manifestation of MEN2A and most frequently occurred along with RET-C634 variant. To facilitate the recognition of MEN2A-CLA, to combine family investigation and screening of RET variant are helpful for early diagnosis and standardized treatment, which can improve the long-term outcome of MEN2A-specific tumors.
Adolescent ; Adrenal Gland Neoplasms ; Adult ; Amyloidosis, Familial ; Carcinoma, Neuroendocrine ; China ; Female ; Humans ; Lichens ; Male ; Middle Aged ; Multiple Endocrine Neoplasia Type 2a/genetics* ; Pheochromocytoma ; Proto-Oncogene Proteins c-ret/genetics* ; Skin Diseases, Genetic ; Thyroid Neoplasms/genetics* ; Young Adult

Objective To indentify the cognitive status of Chinese patients to acne and the influencing factors to theirs' cognitive status,so as to provide solid evidences for the prevention and treatment of acne.Methods A self-designed questionnaire was made to conduct this survey of 16,156 acne patients,who seeked to the treatment in the dermatological departments from 112 hospitals in China.The survey consisted of several parts,including the general status of patients,the patients' cognition of occurrence,development and risk factors of acne,whether the first choice was seeking treatment at the hospital when the patients had acne and the condition of selection of skin care products.The factors were analyzed,which could impact the cognition of the patients' behavior of treatment,how did the patients' cognition to influence their medical behavior and skin care as well as the consistency of assessment of the severity of acne by doctors and patients themselves.Results The acne patients studied had the best knowledge of "acne is a skin disease","it not only occurs in the period of adolescence" and "the disease can be prevented and cured",which accordingly accounted for 80.65％,69.16％ and 65.49％ of the total patients respectively.However,the awareness of acne patients to heredity,high sugar and dairy products as risk factors for acne was insufficient,which accounted for 48.72％,42.40％ and 18.25％ of the total patients,respectively.Gender,age,educational level,occupation and health knowledge were the main factors affecting the cognitive level of patients;the survey also found that men,patient with educational level of junior high or even lower educational condition,occupation of labor workers or farmers and patients were lack of health education with poor knowledge of the genetics and dietary were risk factors for acne;patients with age over 36 years or with mild illness had poor knowledge of dietary risk factors for acne;the difference was statistically significant (P＜0.05).The analysis of the influence of cognitive status on medical treatment behavior and skin care showed that the better the cognition,the higher the probability of patients would choose medical treatment as the first choice as well as choosing functional skin care products;the difference was statistically significant (P＜0.05).The consistency of assessment of the severity of acne by doctors and patients was poor (Kappa value ＜0.4),and the assessment of severity of acne by patients was more serious than doctors' assessment.Conclusions Patient's cognitive status will affect their medical behavior and skin care,and there is also a phenomenon that patients have a more serious assessment of their acne condition.It is suggested that health education for acne patients should be strengthened in clinical medicine so as to improve their knowledge of acne as well as preventing from acne effectively.

Objective Among the factors affecting embryo implantation, oxidative stress is one that receives much medical attention. The purpose of the study was to explore the effect of H2 O2 on the endometrial stromal cells ( ESCs) in the decidual mouse model of oxidative stress of ESCs. Methods Primary ESCs cultured by enzymatic digestion with a mesh filter were divided into a control, an experimental, and a model group. Decidualization was induced in the mice of the experimental and model groups by inter-vention with 10 nM E2 and 1μmol/L P4 for 72 h in vitro. Then, the animals of the model group were treated with different concentra-tions of H2 O2 for 4 hours. The primary ESCs were identified by immunohistochemistry, the expression of dPRP mRNA determined byRT-PCR, the proliferation of the decidual ESCs treated with H2 O2 analyzed by CCK-8, and the level of ROS detected by flow cytometry. Results Primary mouse ESCs were successfully isolated, cultured, and identified, which were shaped like spindles and polygons, radial-ly aligned under the microscope. Immunofluorescence analysis showed positive expression of vimentin and negative expression of cytokeratin. The purity of the primary mouse ESCs was ( 96. 3 ± 0. 49 )%. Theexpression of dPRP mRNA was significantly higher in the ESCs treated with E2 and P4 than in the control (0.0002±0.0000 vs 1.0010±0.0011, P<0.01). H2O2 at ≥150μmol/L suppressed the proliferation of the decidual ESCs by 6.9% (P<0.05), and in a concen-tration-dependent manner, reaching the maximum inhibition rate of 70.6% at the concentration of 300μmol/L. The level of intracellular ROS was markedly increased in the ESCs treated with H2O2 at 50μmol/L (27.77±4.20) and 100μmol/L (43.57±6.58), with statisti-cally significant difference from that in the control group (17.47±0.61) (P=0.001). Conclusion H2O2 at 100μmol/L can signifi-cantly elevate the intracellar ROS level without affecting the proliferation of decidual primary ESCs, and therefore can be used for estab-lishing the model of oxidative stress of ESCs in decidual mice.

Objective To understand the abnormal status of alanine aminotransferase (ALT),fasting blood glucose (GLU),to-tal cholesterol (TC)and triglyceride (TG)in urban and rural residents,to provide the basis for making the effective prevention measures,health education and health promotion to community work more targeted and effective.Methods The data of physical ex-amination results of 4 042 residents were analyzed retrospectively,the levels and abnormal rates of ALT,TG,TC,GLU were com-pared between different gender and age.Results Among 4 042 residents,the abnormal rate of male was significant higher than that of female,but the abnormal rates of TG,TC of female were significant higher than those of male(P <0.05).The highest rates of ALT,TG,TC,GLU in male were in 20-<40 years old,40-<60 years old,60-<80 years old,≥80 years old.The highest rates of ALT,TG,TC,GLU in female were in 0-<20 years old,60 -<80 years old,≥ 80 years old,≥80years old.Conclusion The health status of urban and rural residents is not optimistic,the community intervention,health guidance should be strengthened to prevent and control the occurrence and development of diseases and improve the health level of urban and rural residents.