OBJECTIVE To investigate the inhibitory effects of Bruceine A(BA)on colon cancer and its underlying mechanisms.METHODS Human colon cancer HT-29 and HCT116 cells were treated with various concentrations of BA(0,1,2,5,10,20,40,80 μmol·L-1).Cell viability was assessed using the Cell Counting Kit-8(CCK-8).Flow cytometry,wound healing assays,and Transwell assays were employed to evaluate the effects of BA on cell apoptosis,cell cycle,invasion,and migration.Mo-lecular docking simulations were used to assess the binding of BA to GSK-3β protein,and Western blot analysis was used to examine protein expression related to the cell cycle,epithelial-mesenchymal transition,and the Wnt/β-catenin signaling pathway.An HT-29 cell subcutaneous xenograft mouse model was established.After tumor formation,mice were randomly divided into three groups(six mice per group):a blank group,a low-dose BA group(0.1 mg·kg-1),and a high-dose BA group(0.2 mg·kg-1).Mice were ad-ministered the drug for 19 d,then sacrificed,and tumor tissues were collected.Tumor volume changes over time were observed;Ki67 immunohistochemistry was used to assess cell proliferation in tumor tissues;Western blot analysis of Wnt/β-catenin signaling pathway protein expression was conducted.RESULTS Compared with the blank group,BA could significantly inhibit the proliferation of HT-29 and HCT116 cells,with IC50 values of 10.80 μmol·L-1 and 17.96 μmol·L-1,respectively.Flow cytometry results showed that BA significantly induced apoptosis of HT-29 cells(P<0.01,P<0.001),and arrested the cell cycle at the S phase,accompanied by de-creased expression of cycle-related proteins CDK2 and Cyclin A(P<0.05,P<0.01,P<0.001).BA inhibited cell migration and in-vasion ability(P<0.05,P<0.01,P<0.001),reduced the expression of EMT-related proteins Snail,Vimentin,and N-Cadherin(P<0.01,P<0.001),and upregulated the expression of E-Cadherin protein.In addition,BA inhibited the expression of β-catenin and p-GSK3β proteins.Wnt agonist LiCl could significantly antagonize the anti-colon cancer effect of BA;Wnt inhibitor XAV939 could enhance the anti-colon cancer effect of BA.In the in vivo experiment,compared with the blank group,the tumor volume of the low-dose and high-dose BA groups was significantly reduced(P<0.05,P<0.001).Immunohistochemistry results showed that compared with the blank group,the expression of Ki67 in tumor tissues of the low-dose and high-dose BA groups was significantly reduced(P<0.001).Western blot results further proved that BA inhibited the Wnt/β-catenin signaling pathway.CONCLUSION BA inhibits the viability,invasion,and migration of colon cancer HT-29 cells,induces apoptosis,and causes cell cycle arrest.Additionally,it significantly suppresses the growth of subcutaneous HT-29 cell xenografts in vivo,possibly related to the Wnt/β-catenin signaling pathway.

Objective:To investigate the different expression levels of programmed cell death 1 ligand 1 (PD-L1) in non-small cell carcinoma (NSCLC) of distribution characteristics of TCM constitutions and prognosis.Methods:The clinical data of 355 NSCLC patients who had been treated with immune checkpoint inhibitors (ICIs) from January 2019 to June 2023 in the Cancer Medical Center of the First Affiliated Hospital of Hunan University of Chinese Medicine were retrospectively analyzed, and their TCM constitutions were determined. According to the expression level of PD-L1, they were divided into three groups: low expression group (TPS≤1%), medium expression group (1% < TPS < 49%) and high expression group (TPS≥50%). Overall survival (OS) of patients was followed up, and the median OS were compared. Kaplan-Meier method was used to draw survival curves, and Log-rank test was used to compare the difference of survival curves. The independent risk factors of OS were analyzed by COX regression.Results:The distribution of different TCM constitutions showed statistical significance across the three groups ( P<0.05). The median OS for the medium and high expression groups were 21.082 months and 25.714 months, respectively, both significantly higher than the 14.437 months for the low expression group ( P<0.05). The survival curve of TCM constitutions showed that the constitutions significantly correlated with the prognosis of ICIs treatment were qi deficiency, phlegm dampness, and blood stasis ( P<0.05 or P<0.01). The median OS from high to low was 44.971 months for phlegm-dampness constitution, 23.297 months for qi-deficiency constitution, and 11.763 months for blood-stasis constitution. COX regression analysis indicated that medium PD-L1 expression ( HR=0.622, 95% CI=0.459,0.844, P=0.002), high PD-L1 expression ( HR=0.509, 95% CI=0.361,0.718, P<0.001), phlegm-dampness constitution ( HR=0.556, 95% CI=0.335,0.924, P=0.024), and blood-stasis constitution ( HR=2.952, 95% CI=1.929,4.518, P<0.001) were independent prognostic factors. Conclusions:The higher the expression level of PD-L1 in NSCLC patients, the better the prognosis of ICIs treatment. The prognosis of ICIs treatment is better for people with phlegm-dampness constitution and poor for those with blood stasis constitution.

OBJECTIVE To investigate the inhibitory effects of Bruceine A(BA)on colon cancer and its underlying mechanisms.METHODS Human colon cancer HT-29 and HCT116 cells were treated with various concentrations of BA(0,1,2,5,10,20,40,80 μmol·L-1).Cell viability was assessed using the Cell Counting Kit-8(CCK-8).Flow cytometry,wound healing assays,and Transwell assays were employed to evaluate the effects of BA on cell apoptosis,cell cycle,invasion,and migration.Mo-lecular docking simulations were used to assess the binding of BA to GSK-3β protein,and Western blot analysis was used to examine protein expression related to the cell cycle,epithelial-mesenchymal transition,and the Wnt/β-catenin signaling pathway.An HT-29 cell subcutaneous xenograft mouse model was established.After tumor formation,mice were randomly divided into three groups(six mice per group):a blank group,a low-dose BA group(0.1 mg·kg-1),and a high-dose BA group(0.2 mg·kg-1).Mice were ad-ministered the drug for 19 d,then sacrificed,and tumor tissues were collected.Tumor volume changes over time were observed;Ki67 immunohistochemistry was used to assess cell proliferation in tumor tissues;Western blot analysis of Wnt/β-catenin signaling pathway protein expression was conducted.RESULTS Compared with the blank group,BA could significantly inhibit the proliferation of HT-29 and HCT116 cells,with IC50 values of 10.80 μmol·L-1 and 17.96 μmol·L-1,respectively.Flow cytometry results showed that BA significantly induced apoptosis of HT-29 cells(P<0.01,P<0.001),and arrested the cell cycle at the S phase,accompanied by de-creased expression of cycle-related proteins CDK2 and Cyclin A(P<0.05,P<0.01,P<0.001).BA inhibited cell migration and in-vasion ability(P<0.05,P<0.01,P<0.001),reduced the expression of EMT-related proteins Snail,Vimentin,and N-Cadherin(P<0.01,P<0.001),and upregulated the expression of E-Cadherin protein.In addition,BA inhibited the expression of β-catenin and p-GSK3β proteins.Wnt agonist LiCl could significantly antagonize the anti-colon cancer effect of BA;Wnt inhibitor XAV939 could enhance the anti-colon cancer effect of BA.In the in vivo experiment,compared with the blank group,the tumor volume of the low-dose and high-dose BA groups was significantly reduced(P<0.05,P<0.001).Immunohistochemistry results showed that compared with the blank group,the expression of Ki67 in tumor tissues of the low-dose and high-dose BA groups was significantly reduced(P<0.001).Western blot results further proved that BA inhibited the Wnt/β-catenin signaling pathway.CONCLUSION BA inhibits the viability,invasion,and migration of colon cancer HT-29 cells,induces apoptosis,and causes cell cycle arrest.Additionally,it significantly suppresses the growth of subcutaneous HT-29 cell xenografts in vivo,possibly related to the Wnt/β-catenin signaling pathway.

Objective To examine the short-term prognostic value of procalcitonin ( PCT ) combined with coagulation factors for cirrhotic patients complicated with spontaneous bacterial peritonitis (SBP).Methods Clinical data of 128 cirrhotic patients complicated with SBP admitted in Jinhua Central Hospital from June 2014 to October 2017 were retrospectively analyzed .In 3 months after admission , 83 patients survived ( survival group ) and 45 patients died ( fatal group ) .The factors related to prognosis were analyzed with Logistic regression and the prediction model was constructed with the weights derived from regression coefficients.The ROC curve and the area under the curve (AUC) of combination of PCT with coagulation factors were used to predict the survival of patients .Results Univariate analysis indicated that the level of PCT , total bilirubin ( TBil ) , serum creatinine ( Scr ) , prothrombin time ( PT ) , prothrombin activity ( PTA ) , blood coagulation factor Ⅱ, Ⅴ, Ⅶ, Ⅸ, Ⅹ, Ⅺ and Ⅻ were factors affecting the prognosis of cirrhotic patients complicated with SBP (P<0.01).Multivariate analysis showed that PCT , blood coagulation factors Ⅴ and Ⅸ were independent factors of short-term prognosis of cirrhotic patients complicated with SBP.The constructed predictive model was Logit (P) =1.200+0.099 ×PCT-0.026 × clotting factor Ⅴ-0.038 ×clotting factor Ⅸ.The sensitivity and specificity of the model were 0.822 and 0.675, respectively, and the AUC was 0.829.Compared with the classic MELD score , the difference was not statistically significant (P>0.05).Conclusions The predictive model based on PCT and coagulation factors Ⅴand Ⅸcan effectively predict the short-term survival of cirrhotic patients complicated with SBP . The overall prognostic ability is not different from MELD score , but the model is more simple and easier to apply.

Background: Bone metastases are common in patients with advanced cancer. Bisphosphonates (BPs) could prevent or delay the development of skeleton-related events (SREs). The present study aimed to identify the clinical features of and treatment strategies for Chinese patients with bone metastases. Methods: Consecutive cancer patients who had bone metastases and received BP treatment were enrolled. A ques-tionnaire was developed to collect the patients’clinical data, as well as information on the diagnosis and manage-ment of bone metastases. Physicians’awareness of the guidelines and knowledge of the application of BP were also assessed. Results: A total of 3223 patients with lung cancer (36.5%), breast cancer (30.9%), prostate cancer (8.5%), and gas-trointestinal cancer (5.7%) were included in this study. The sites of bone metastases were the thoracic spine (56.0 %), lumbar spine (47.1%), ribs (32.6%), and pelvis (23.2%). The SRE frequency was the highest in patients with multiple myeloma (36.6%), followed by those with lung cancer (25.9%), breast cancer (20.2%), prostate cancer (18.2%), and gas-trointestinal cancer (17.3%). Irradiation to the bone was the most frequent SRE (58% in lung cancer patients, 45% in breast cancer patients, and 48% in prostate cancer patients). Our survey also showed that 45.5% of patients received BP within 3 months after their diagnosis of bone metastases, whereas the remaining 54.5% of patients did not receive BP treatment until at least 3 months after their diagnosis of bone metastases. The SRE frequency in the former group was significantly lower than that in the latter group (4.0% vs. 42.3%, P < 0.05). In patients with more than 6 months of continuous BP treatment, the mean time to the first SRE was significantly longer than that in patients with less than 6 months of continuous BP treatment (7.2 vs. 3.4 months, P < 0.05). In addition, 12.2% of the physicians were not aware of the efcacy of BP in preventing and delaying SRE. Only half (52.3%) of the physicians agreed that the BP treatment should persist for at least 6 months unless it was intolerable. Conclusions: Our study suggested that prompt and persistent BP treatment was associated with a reduced risk of SREs. However, our survey also revealed that the proper application of BP was not as common as expected in China.

Objective To observe the effect of transforming growth factor-β1 (TGF-β1) vaccine on the degree of hepatic fibrosis in rats ,and to explore the effect of TGF-β1 vaccine on the insulin-like growth factor binding protein (IGFBP)3 and IGFBP7 .Methods The hepatic fibrosis rat model was set up by injecting N-nitrosodimethylamine . Among them , 10 rats were injected with TGF-β1 vaccine , and additional 10 rats were set up as healthy control group .Changes in hepatic pathology were observe and the expressions of IGFBP3 and IGFBP7 were detected by the methods of immunohistochemistry , reverse transcription polymerase chain reaction (RT-PCR) and Western blot in rat fibrosis tissues after 6 weeks . Normality test and analysis of variance were conducted .LSD test was conducted if variances were tested homogeneity .Categorical data were analyzed using Fisher exact test . Results Changes in hepatic histology and serum levels of hyaluronic acid and laminin suggested that TGF-β1 vaccine interventions could reduce the extent of hepatic fibrosis in rats .The expressions of IGFBP3 mRNA in control group ,hepatic fibrosis model group and vaccine intervention group were 1 .735 ± 0 .097 ,1 .165 ± 0 .096 and 1 .491 ± 0 .046 ,respectively (t= 4 .575 ,6 .285 and 8 .489 ,respectively ,all P< 0 .05) .The expressions of IGFBP7 in the above three groups were 0 .497 ± 0 .021 ,1 .250 ± 0 .064 and 0 .885 ± 0 .149 ,respectively (t= 5 .161 ,30 .101 and 7 .250 , respectively ,all P < 0 .05 ) . Immunohistochemistry proved that the expressions of IGFBP7 in fibrosis model group and TGF-β1 vaccine group were all significantly higher than control group ;and the expressions of IGFBP3 in fibrosis model group and TGF-β1 vaccine group were all significantly lower than control group .The expressions of IGFBP3 protein in control group , hepatic fibrosis model group and vaccine intervention group were 7 .508 ± 0 .357 ,5 .200 ± 0 .210 and 5 .751 ± 0 .178 ,respectively (t = 7 .622 ,6 .180 and 29 .156 , respectively ,all P < 0 .05) . The expressions of IGFBP7 were 1 .176 ± 0 .051 ,1 .735 ± 0 .115 and 1 .428 ± 0 .056 ,respectively (t = 7 .188 ,4 .827 and 8 .649 ,respectively ,all P< 0 .05) .Conclusion TGF-β1 vaccine can affect the expressions of IGFBP3 and IGFBP7 ,which plays an important role in the formation and development of hepatic fibrosis .

ObjectiveTo evaluate the efficacy about the different wearing manner with Milwaukee and Boston for the treatment of adolescent idiopathic scoliosis (AIS).MethodsRetrospectively summarization and analysis was performed in 85 adolescent patients with idiopathic scoliosis who were treated from February 2004 to March 2009.The skeletal growth of them had not completed.There were 57 cases who received brace treatment.In them,28 were treated with (CTLSO) Milwaukee brace,and the rest were treated with (TLSO) Boston brace and orthopedic gymnastics.The 30 cases wear regularly used for 21 to 23hours per day,and 27 cases could consist on wearing only 6 to 15 hours per day.The other 28 cases of 85AIS cases were only treated with orthopedic gymnastics instead of orthosis treatment.All patients were periodically observed with lateral side (X)-ray photograph at standing position and photograph,and Cobb angle and Risser sign were measured every 3 to 6 months.Since 2008 all adolescent idiopathic scoliosis patients treated with orthosis were requested to fill with simplified Chinese SRS-22.Results73 cases adolescent idiopathic scoliosis patients were followed up for 2 ～5 years [ mean(26.3 ± 33.7)months ].Milwaukee orthosis group showed the regular wearing group had 91.67 ％ ( 11/12) effective rate and the intermittent wearing group had 56.25％ (9/16) effective rate and the group without wearing orthosis only had 20％ (4/20)effective rate.The group regularly wearing Milwaukee brace had superior effect than the other two groups (P ＜ 0.05 ).Boston orthosis group showed the regular wearing group had 88.89％ (16/18) effective rate and the intermittent wearing group had 54.55％ (6/11 ) effective rate and the group without wearing brace with 25％ (2/8) effective rate.The group regularly wearing Boston brace had better effect than the other two groups ( P ＜ 0.05 ).Due to the different choice of AIS patients and orthosis,the effective rate of the Milwaukee and Boston orthosis was not compared.ConclusionsThe adolescent idiopathic scoliosis patients should insist on regularly wearing brace regardless of the Milwaukee or Boston orthosis ( this article suggest that the wearing time should not less than 21 ～ 23 h/d).The group regularly wearing with the Milwaukee or Boston orthosis had better effect than the intermittent group or the group without wearing brace.It's a good treatment for the AIS patients who have with the indication of orthosis treatment.

ObjectiveTo find out the risk factors causing iatrogenic spinal cord injury (ISCI) so as to provide theoretical support for reducing the spinal cord injury during spinal operation. Methods A retrospective study was done on 120 patients undergone cervical or thoracic spinal( C1-T12 ) surgery at Xiangya Hospital of Central South University from January 2002 to January 2009. The patients were randomly divided into injury group (n = 34) and control group (n = 86) and the univariate analysis was used to analyze 30 factors including clinical factors, iconography factors, operation and pathology factors as well as possible protective factors. Then, the factors with statistical difference were analyzed by using the multi-factor unconditioned Logistic analysis.Results The univariate comparison between the two groups showed statistical difference ( P ＜ 0. 05 ) in nine factors including combined hypertension, combined diabetes mellitus, preoperative ASIA grade, spinal canal stenosis rate, ratio of spinal cord area/efficient area of vertebral canal, spinal cord MRI T2WI high signal, bleeding amount during operation, intraspinal prominence adhesion to dura mate of spinal cord as well as intraoperative use of methylprednisolone. The multi-factor Logistic regression analysis revealed that ASIA grade, value of spinal cord area/efficient area of vertebral canal, spinal cord MRI T2W1 high signal and bleeding amount in operation had positive correlation with ISCI. Use of methylprednisolone during operation had negative correlation with ISCI. ConclusionsCombined diabetes mellitus, ASIA grade, spinal cord MRI T2W1 high signal, ratio of spinal cord/vertebral canal area and bleeding amount in operation are the risk factors for ISCI. Use of large dose methylprednisolone exerts preventive effect on ISCI.