1.Effects of FAK phosphorylation on adhesion and migration of smooth muscle cells stimulated by fibronectin
Hang YIN ; Lihui WANG ; Xu PENG ; Yong HUO ; Chunfang XIA ; Chaoshu TANG
Chinese Journal of Pathophysiology 1989;0(06):-
AIM: To study the effect of focal adhesion kinase (FAK) phosphorylation on the adhesion and migration of smooth muscle cells (SMCs) stimulated by fibronectin (FN). METHODS: Cultured SMCs were stimulated by different concentrations of FN.FAK expression and phosphorylation were detected by immunoprecipitation and Western blot. To investigate the modulating effect of FAK on tyrosine phosphorylation and SMCs adhesion and migration, FAK antisense oligodeoxynucleotides (ODNs) were transfected into SMCs by actionic lipid method. RESULTS: FAK expression increased when SMCs adhesion and migration were induced by FN at different concentrations used in the experiment. However, FAK phosphorylation was only observed by FN stimulation at concentration of 20 mg/L. FAK antisense ODNs inhibited FAK phosphorylation magnificently. The migration rates of SMCs were reduced by 17.89%-27.67% when FN was used at concentration from 5 mg/L to 60 mg/L. The decreased migrating cell numbers were showed the same patterns. The apoptotic SMCs were 33.57% higher than that control ( P
2.Phenotypic modulation of vascular smooth muscle cells and expression of p38 mitogen-activated protein kinase after intimal injury of rabbit carotid arteries
Xinping ZHANG ; Yong HE ; Yuehua PANG ; Xiaoli XIA ; Hang LU ; Gelan BIE ; Chaoxin ZHANG ; Xuelian ZHANG ; Qing LV
Journal of Chinese Physician 2000;0(12):-
Objective To explore phenotypic modulation of vascular smooth muscle cells(VSMC) and change of p38 mitogen-activated protein kinase(MAPK) expression after intimal injury of rabbit carotid arteries. Methods The model of vascular restenosis established by balloon injury of rabbit carotid common arteries was used.HE staining,immunohistochemistry staining and Western blot were used to detect the change of proliferation cell nuclear antigen(PCNA),smooth muscle ?-actin(SM?-actin),p38 expression and morphology of sham-injured arteries and injured arteries at different time points.Results ⑴The proliferating VSMC was observed on the side of the medium lumen at 1 day and on the surface of vascular lumen at 3 days after injury.The neointima was formed and gradually being thicken at 5~7 days,and the thickening was accelerated at 14~35 days after injury.⑵PCNA was negative in the medium and endothelium of sham-injured arteries.Positive cell rate of PCNA was gradually increased at 1~14 days in the medium and at 5~14 days in the neointima after injury,with the maximum rate at 14 days.However,it declined gradually after 28 days.Positive cell rate of PCNA in the neointima was slightly higher than that in the medium.⑶SM?-actin was positive in the medium,negative in the endothelium of sham-injured arteries.Positive cell area of SM?-actin initially decreased at 1 day in the medium after injury with the minimum rate at 3 days,but it increased gradually after 5 days.Expression of SM?-actin in the neointima was slightly less than that in the medium.⑷p38 expression was very low or negative in the medium of sham-injured arteries.Expression of p38 was sustained and increased at 1~35 days after injury with the most remarkable elevation at 3~14 days.Expression of p38 in the neointima was higher than that in the medium.There was positive relationship between the p38 expression and PCNA expression in the vascular wall at different time points after injury.Elevation of p38 was earlier than decrease of SM?-actin.Conclusion There was a close relationship between the phenotypic modulation and proliferating ability of VSMC.P38 participated in signal transduction of phenotypic modulation of VSMC after intimal injury.
3.FAK antisense oligodeoxynucleotides inhibit vascular smooth muscle cell migration and adhesion mediated by FAK-ERK1/2 signaling pathway.
Hang YIN ; Li-hui WANG ; Yong HUO ; Xu PENG ; Chun-fang XIA ; Chao-shu TANG
Acta Pharmaceutica Sinica 2002;37(5):334-338
AIMTo study the effects of FAK-ERK1/2 signaling pathway and FAK antisense oligodeoxynucleotides (ODNs) on vascular smooth muscle cell (SMC) migration and adhesion stimulated by fibronectin (FN).
METHODSMigration and adhesion of cultured SMCs were stimulated by different concentrations of FN, FAK, ERK1/2. And their phosphorylation were detected by immunoprecipitation and Western blot. FAK antisense ODNs were transfected into SMCs by cationic lipid to investigate its modulatory effects on tyrosine phosphorylation, SMCs migration and adhesion were also measured by modifing Boyden Chamber and morphological enumeration, respectively.
RESULTSFAK were expressed when SMCs adhesion and migration were successfully simulated by FN (5, 10, 20, 40, 60 micrograms.mL-1), high contents of FAK and ERK1/2 phosphorylation were detected by 20 micrograms.mL-1 FN or more. FAK antisense ODNs were transfected efficiently by cationic lipid. FAK and ERK1/2 phosphorylation were inhibited magnificently after FAK antisense ODNs transfection. Cell migration stimulated by FN 10, 20, 40 and 60 micrograms.mL-1 were reduced by 23.26%, 21.63%, 19.31% and 17.88% respectively (P < 0.05). SMCs adhesive spreading in 5-60 micrograms.mL-1 FN groups were reduced by 17.89%-27.67% (P < 0.05).
CONCLUSIONFAK-ERK1/2 mediated signal transduction play important roles in SMCs migration and adhesion stimulated by extracellular matrix. The process can be inhibited by FAK antisense ODNs effectively.
Animals ; Aorta ; cytology ; Cell Adhesion ; drug effects ; Cell Movement ; drug effects ; Cells, Cultured ; Fibronectins ; pharmacology ; Focal Adhesion Kinase 1 ; Focal Adhesion Protein-Tyrosine Kinases ; Mitogen-Activated Protein Kinase 1 ; biosynthesis ; metabolism ; Mitogen-Activated Protein Kinase 3 ; Mitogen-Activated Protein Kinases ; biosynthesis ; metabolism ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Oligodeoxyribonucleotides, Antisense ; pharmacology ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; genetics ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Transfection
4.Activated focal adhesion kinase involved in adhesion and migration of vascular smooth muscle cells stimulated by fibronectin.
Hang YIN ; Lihui WANG ; Yong HUO ; Xu PENG ; Chunfang XIA ; Chaoshu TANG
Chinese Medical Journal 2002;115(4):494-497
OBJECTIVETo study the effects of focal adhesion kinase (FAK) phosphorylation on smooth muscle cells (SMCs) adhesion and migration stimulated by fibronectin.
METHODSAdhesion and migration of cultured SMCs were stimulated by different concentrations of fibronectin (FN), FAK and its phosphorylation were detected by immunoprecipitation and Western blot. FAK antisense oligodeoxynucleotides (ODNs) were transfected into SMCs by cationic lipid to investigate its modulatory effects on tyrosine phosphorylation. SMCs adhesion and migration were also measured by morphological enumeration and modified Boyden Chambers, respectively.
RESULTSFAK were expressed when SMCs adhesion and migration were successfully simulated by different concentrations of FN. FAK phosphorylation were detected only at 20 microg/ml FN or more. FAK antisense ODNs were transfected efficiently by cationic lipid and FAK phosphorylation was inhibited substantially. The SMCs migration rate in the 5 - 60 microg/ml FN groups was reduced by 17.89% - 27.67%. Cell migration stimulated by FN at 10, 20, 40 and 60 microg/ml were reduced by 23.26%, 21.63%, 19.31% and 17.88%, respectively (P < 0.05).
CONCLUSIONSFAK phosphorylation and FAK-mediated signal transduction play important roles in SMCs adhesion and migration stimulated by ECM. The process can be inhibited effectively by FAK antisense ODNs.
Animals ; Cell Adhesion ; drug effects ; Cell Movement ; drug effects ; Cells, Cultured ; DNA, Antisense ; genetics ; physiology ; Dose-Response Relationship, Drug ; Enzyme Activation ; drug effects ; Fibronectins ; pharmacology ; Focal Adhesion Kinase 1 ; Focal Adhesion Protein-Tyrosine Kinases ; Muscle, Smooth, Vascular ; cytology ; drug effects ; Phosphorylation ; drug effects ; Protein-Tyrosine Kinases ; genetics ; metabolism ; Rats ; Transfection
5.Multi-center study of motherwort injection to prevent postpartum hemorrhage after caesarian section
Jianhua LIN ; Qide LIN ; Xinghui LIU ; Jianying YAN ; Jing HE ; Li LI ; Hang GU ; Lizhou SUN ; Jianping ZHANG ; Song YU ; Yuyan MA ; Jianmin NIU ; Yong XIA ; Sancun ZHAO ; Wang LI ; Huilan WANG ; Bingshun WANG
Chinese Journal of Obstetrics and Gynecology 2009;44(3):175-178
Objective To evaluate the efficacy and safety of motherwort (herbs leonuri/leonurus heterophyllus sweet) injection for preventing postpartum hemorrhage after caesarian section. Methods The prospective study was designed as a randomized and single blind multi-center research matched with positive agent as controls from Apt 2007 to Aug 2007. 440 women underwent caesarian section (CS) indicated by obstetric factors were enrolled from 15 teaching hospitals in China and assigned into three groups: group of motherwort: 147 cases were administered by motherwort 40 rag uterine injection during CS and 20 mg intramuscular injection per 12 hours 3 times after CS; group of motherwort + oxytocin : 144 cases were administered by motherwort 40 mg and oxytocin 10 U uterine injection during CS and motherwort 20 mg intramuscular injection per 12 hours 3 times after CS and group of oxytocin: 149 cases were administered by oxytocin 10 U uterine injection and oxytocin 10 U + 5% glucose 500 nd intravenously injection during operation and oxytocin 10 U intramuscular injection per 12 hours 3 times after CS. The following clinical parameter were collected and analyzed: (1) The amount of blood loss during operation, at 2, 6, 12, 24, 48 hours after operation. (2) The total amount of blood loss in 24 hours after CS and the incidence of postpartum hemorrhage. (3) The change of level of hemoglobin (Hb) and counting of red blood cell ( RBC ) from prepartum to postpartum. (4) Adverse reaction. Results (1) The mean amount of blood loss during operation were (368±258) ml in group of motherwort, (255±114) mi in group of motherwort + oxytocinand (269±141 ) ml in group of oxytocin, which exhibited significant difference among three groups ( P<0.01 ). Meanwhile, no statistical different amount of blood loss among three groups were observed at 2,6,12, 24, 48 hours after CS. (2)The amount of blood loss of postpartum at 24 hours were (480±276)ml ingroup of motherwort, (361±179) ml in group of motherwort + oxytocin, (381±179) nd in group of oxytocin, which showed significant difference among 3 groups(P <0.01 ). (3) The incidence of postpartum hemorrhage were 32.0% (47/147) in group of motherwort, 11.1% (16/144) in group of motherwort + oxytocin, and 18.8% in (28/149) in group of oxytocin. When comparing the lowest rate of postpartum blood loss in group of motherwort + oxytocin and the highest rate in group of motherwort, it displayed statistical difference (P<0.01). (4) The decreased level of RBC and Hb were shown that RBC(0.3±0.5)×10<'12<‘/L and Hb(9±13)g/L in group of motherwort, RBC (0.2±0.4)×10<'12/L and Hb ( 6±10) g/Lin group of motherwort + oxytocin and RBC (0.2±0.4)×10<'12/L and Hb(7±30) g/L in group of oxytocinrespectively. However, the comparison of different value of RBC and lib in group of oxytocin and motherwort +oxytocin showed significant difference (P<0.05 ). (5) Two cases with allery reaction was observed.Conclusion It is safe and efficacious that combined use of motherwort injection and oxytocin was to preventpostpartum hemorrhage during or after caesarian section.
6.Construction of anti-cD20scFv/CD80/CD28/zeta recombinant gene modified T cell and research on its targeting cytotoxicity.
Yong-Xian HU ; Kang YU ; Ying-Xia TAN ; Zhi-Jian SHEN ; Song-Fu JIANG ; Hong-Lan QIAN ; Bin HANG ; Da-Ming SHAN
Chinese Journal of Hematology 2007;28(2):111-114
OBJECTIVETo construct anti-CD20scFv/CD80/CD28/zeta recombinant gene modified T cells, test its effectiveness of eradicating CD20+ lymphoma cells and provide a probably new approach to tumor adoptive immunotherapy.
METHODSCD28-zeta cDNA were amplified from vector pBULLET and inserted into pLNCX vector that contained anti-CD20scFv/CD80 gene. The recombinant vectors were transduced into PA317 cells and high titer retroviruses were obtained to infect human peripheral blood T lymphocytes. Resistant T cells were obtained by G418 selection at one week. Then transduced T lymphocytes and lymphoma cell lines Daudi Raji were cocultured. The cytotoxicity and cytokine production of transduced T cells were determined by non-radio-activation cytotoxicity assay and ELISA respectively.
RESULTSThe recombinant eukaryotic vector was constructed successfully as proved by enzyme digestion analysis and sequencing. These T cells were able to lyse CD20+ target cells and secrete high levels of IL-2 and IFN-gamma in vitro.
CONCLUSIONRecombinant gene modified T cells can be constructed successfully. It can specially kill CD20 positive lymphoma cells in vitro.
Antigens, CD20 ; genetics ; immunology ; B7-1 Antigen ; genetics ; immunology ; CD28 Antigens ; genetics ; immunology ; Cell Line ; Cytotoxicity, Immunologic ; Genetic Vectors ; Humans ; Immunotherapy, Adoptive ; Plasmids ; genetics ; T-Lymphocytes ; immunology ; metabolism ; Transfection
7.Analysis of timing and prognostic factors of early tracheotomy in patients with multiple rib fractures
Bing ZHANG ; Gongke LI ; Yurong WANG ; Fei WU ; Suqin SHI ; Qinling FENG ; Xin HANG ; Runfeng MIAO ; Le XIA ; Cheng DUAN ; Juling LENG ; Yong LI
Chinese Journal of Trauma 2021;37(7):646-652
Objective:To investigate the related factors that affect the timing and prognosis of early tracheostomy in patients with multiple rib fractures.Methods:A retrospective case series study was conducted on medical data of 222 patients with multiple rib fractures who underwent tracheostomy in Affiliated Hospital of Yangzhou University from February 2013 to October 2019,including 160 males and 66 females,with the age of 18 to 85 years [(49.5 ± 16.3)years]. According to the practice management guidelines for tracheostomy timing and the use of propensity score matching technology,there were 118 patients with tracheostomy within 7 days of tracheal intubation (early group) and 104 patients with tracheostomy after 7 days of tracheal intubation (late group) before matching,and there were 87 patients in early group and 87 patients in late group after matching. Data were compared between groups including the gender,age,underlying disease,injury severity score (ISS),Glasgow coma score (GCS),number of fractured ribs,total number of rib fractures (NTRF),first rib fracture,flail chest,traumatic brain injury,combined injuries (spine,maxillofacial,sternum),acute respiratory distress syndrome (ARDS),volume fraction of pulmonary contusion(VPC),blood lactic acid (within 24 hours of admission),hemothorax,pneumothorax,mechanical ventilation time,duration of tracheostomy,time from tracheal intubation to incision,length of hospital stay,length of stay in ICU,closed thoracic drainage,number of fiberoptic bronchoscopy,multi-drug resistant bacteria infection,ventilator-associated pneumonia,antibiotic use time,duration of sedative and analgesic drugs used and 28-day mortality. The multivariate Logistic regression analysis was used to predict independent risk factors for early tracheostomy. The Pearson method was used to compare the relationship between multiple factors. The receiver operating characteristic (ROC) curve was used to predict indicators that affect the prognosis of patients with early tracheostomy,and calculate the best cut-off value. The Kaplan-Meier single factor and COX multivariate survival were used to analyze the relevant factors affecting the 28-day mortality of patients.Results:(1) In early group,the NTRF,ARDS and VPC were higher than those in late group,and the time from tracheal intubation to incision and 28-day mortality rate were lower than those in late group ( P < 0.05),while the two groups showed no significant differences in the gender,age,underlying diseases and ISS ( P > 0.05). (2) The multivariate Logistic regression analysis showed that there was statistical significance in NTRF ( OR = 1.775,95% CI 1.439-2.188),ARDS( OR = 3.740,95% CI 1.441-9.711),VPC ( OR = 1.087,95% CI 1.052-1.124) ( P < 0.05); the Pearson method analysis showed a significant correlation between VPC and NTRF ( r = 0.369, P < 0.05) and a low degree of correlation between ARDS and VPC ( r = 0.179, P < 0.05),but there was no significant correlation between ARDS and NTRF ( r = 0.132, P > 0.05). (3) The ROC curve analysis showed that the area under the curve (AUC) of the VPC and NTRF [AUC = 0.832 (95% CI 0.770-0.893),AUC = 0.804 (95% CI 0.740-0.868)] were significantly higher than those of the number of rib fractures [AUC = 0.437(95% CI 0.352-0.523),GCS [AUC = 0.519 (95% CI 0.432-0.605)] and ISS [AUC = 0.484 (95% CI 0.398-0.571)] ( P < 0.05). After calculating the Yorden index,the best cut-off value for VPC was 23.9,and the best cut-off value for NTRF was 8.5. (4) The Kaplan-Meier single factor and multivariate COX model survival analysis showed that the 28-day survival ratio of patients with early tracheostomy was significantly better than that of late tracheostomy ( P < 0.05). Conclusions:The NTRF,ADRS and VPC are independent risk factors for the timing and prognosis of early tracheostomy. There is a significant correlation between VPC and NTRF. The VPC ≥ 23.9% and or NTRF ≥ 8.5 can be used to predict early tracheostomy in patients with multiple rib fractures. Early tracheostomy may benefit the 28-day survival of patients with multiple rib fractures.
8. Observation on the efficacy of motherwort injection in the treatment of abnormal uterine bleeding-ovulatory dysfunction
Wan-jing LI ; Wan-ting XIA ; Hang ZHOU ; Yong-wen LI ; Jing YUAN ; Xiao-lin XIE ; Fang HE ; Qian ZENG
Chinese Journal of Practical Gynecology and Obstetrics 2019;35(10):1150-1155
OBJECTIVE: To observe the clinical efficacy of motherwort injection in the treatment of abnormal uterine bleeding-ovulatory dysfunction(AUB-O),and provide data support for broadening the clinical application range of motherwort injection.METHODS: A multicenter,randomized,prospective study was used.The patients who were diagnosed with AUB-O in Hospital of Chengdu University of Traditional Chinese Medicine in 2018 were randomly divided into the control group and the experimental group according to random numbers.There were 75 cases in the experimental group,7 cases were lost,and 68 cases were the final cases;another 75 cases were in the control group,4 cases were lost,and there were 71 cases in the end.In the control group,only the tranexamic acid was given.On the basis of this,the experimental group was given intramuscular injection of motherwort injection 2 mL/time,b.i.d.,for 3 consecutive days.The clinical TCM symptom scores,blood loss,blood routine,coagulation function,endometrial thickness and effective rate were compared between the two groups.All data were statistically analyzed using IBM SPSS 21.0.RESULTS: After treatment,the total effective rate of the experimental group was 97.06%,which was significantly better than that(84.51%)of the control group(P<0.05).TCM syndrome scores of Qi stagnation and blood stasis of the experimental group was significantly lower than those in the control group(P<0.05).TCM syndrome scores of damp-heat stagnation of the experimental group was significantly lower than those in the control group(P<0.05).There was no significant difference in the amount of bleeding between the two groups on the first day after treatment(P>0.05),but on the 2nd day and the 3rd day after treatment,the difference was statistically different(P<0.05),and the cumulative amount of bleeding after 3 days of treatment was statistically different(P<0.05).The endometrial thickness of the experimental group was lower than that of the control group(P<0.01).CONCLUSION: Motherwort injection combined with basic therapy is effective in the treatment of AUB-O.
9.Construction of a model based on multipoint full-layer puncture biopsy for predicting pathological complete response after neoadjuvant therapy for locally advanced rectal cancer
Ying JIN ; Zhiwei ZHAI ; Liting SUN ; Pingdian XIA ; Hang HU ; Chongqiang JIANG ; Baocheng ZHAO ; Hao QU ; Qun QIAN ; Yong DAI ; Hongwei YAO ; Zhenjun WANG ; Jiagang HAN
Chinese Journal of Gastrointestinal Surgery 2024;27(4):403-411
Objective:To investigate the value of transanal multipoint full-layer puncture biopsy (TMFP) in predicting pathological complete response (pCR) after neoadjuvant radiotherapy and chemotherapy (nCRT) in patients with locally advanced rectal cancer (LARC) and to establish a predictive model for providing clinical guidance regarding the treatment of LARC.Methods:In this multicenter, prospective, cohort study, we collected data on 110 LARC patients from four hospitals between April 2020 and March 2023: Beijing Chaoyang Hospital of Capital Medical University (50 patients), Beijing Friendship Hospital of Capital Medical University (41 patients), Qilu Hospital of Shandong University (16 patients), and Zhongnan Hospital of Wuhan University (three patients). The patients had all received TMFP after completing standard nCRT. The variables studied included (1) clinicopathological characteristics; (2) clinical complete remission (cCR) and efficacy of TMFP in determining pCR after NCRT in LARC patients; and (3) hospital attended, sex, age, clinical T- and N-stages, distance between the lower margin of the tumor and the anal verge, baseline and post-radiotherapy serum carcinoembryonic antigen (CEA) and carbohydrate antigen (CA)19-9 concentrations, chemotherapy regimen, use of immunosuppressants with or without radiotherapy, radiation therapy dosage, interval between surgery and radiotherapy, surgical procedure, clinical T/N stage after radiotherapy, cCR, pathological results of TMFP, puncture method (endoscopic or percutaneous), and number and timing of punctures. Single-factor and multifactorial logistic regression analysis were used to determine the factors affecting pCR after NCRT in LARC patients. A prediction model was constructed based on the results of multivariat analysis and the performance of this model evaluated by analyzing subject work characteristics (ROC), calibration, and clinical decision-making (DCA) curves. pCR was defined as complete absence of tumor cells on microscopic examination of the surgical specimens of rectal cancer (including lymph node dissection) after NCRT, that is, ypT0+N0. cCR was defined according to the Chinese Neoadjuvant Rectal Cancer Waiting Watch Database Study Collaborative Group criteria after treatment, which specify an absence of ulceration and nodules on endoscopy; negative rectal palpation; no tumor signals on rectal MRI T2 and DWI sequences; normal serum CEA concentrations, and no evidence of recurrence on pelvic computed tomography/magnetic resonance imaging.Results:Of the 110 patients, 45 (40.9%) achieved pCR after nCRT, which was combined with immune checkpoint inhibitors in 34 (30.9%). cCR was diagnosed before puncture in 38 (34.5%) patients, 43 (39.1%) of the punctures being endoscopic. There were no complications of puncture such as enterocutaneous fistulae, vaginal injury, prostatic injury, or presacral bleeding . Only one (2.3%) patient had a small amount of blood in the stools, which was relieved by anal pressure. cCR had a sensitivity of 57.8% (26/45) for determining pCR, specificity of 81.5% (53/65), accuracy of 71.8% (79/110), positive predictive value 68.4% (26/38), and negative predictive value of 73.6% (53/72). In contrast, the sensitivity of TMFP pathology in determining pCR was 100% (45/45), specificity 66.2% (43/65), accuracy 80.0% (88/110), positive predictive value 67.2% (45/67), and negative predictive value 100.0% (43/43). In this study, the sensitivity of TMFP for pCR (100.0% vs. 57.8%, χ 2=24.09, P<0.001) was significantly higher than that for cCR. However, the accuracy of pCR did not differ significantly (80.0% vs. 71.8%, χ 2=2.01, P=0.156). Univariate and multivariate logistic regression analyses showed that a ≥4 cm distance between the lower edge of the tumor and the anal verge (OR=7.84, 95%CI: 1.48-41.45, P=0.015), non-cCR (OR=4.81, 95%CI: 1.39-16.69, P=0.013), and pathological diagnosis by TMFP (OR=114.29, the 95%CI: 11.07-1180.28, P<0.001) were risk factors for pCR after NCRT in LARC patients. Additionally, endoscopic puncture (OR=0.02, 95%CI: 0.05-0.77, P=0.020) was a protective factor for pCR after NCRT in LARC patients. The area under the ROC curve of the established prediction model was 0.934 (95%CI: 0.892-0.977), suggesting that the model has good discrimination. The calibration curve was relatively close to the ideal 45° reference line, indicating that the predicted values of the model were in good agreement with the actual values. A decision-making curve showed that the model had a good net clinical benefit. Conclusion:Our predictive model, which incorporates TMFP, has considerable accuracy in predicting pCR after nCRT in patients with locally advanced rectal cancer. This may provide a basis for more precisely selecting individualized therapy.
10.Construction of a model based on multipoint full-layer puncture biopsy for predicting pathological complete response after neoadjuvant therapy for locally advanced rectal cancer
Ying JIN ; Zhiwei ZHAI ; Liting SUN ; Pingdian XIA ; Hang HU ; Chongqiang JIANG ; Baocheng ZHAO ; Hao QU ; Qun QIAN ; Yong DAI ; Hongwei YAO ; Zhenjun WANG ; Jiagang HAN
Chinese Journal of Gastrointestinal Surgery 2024;27(4):403-411
Objective:To investigate the value of transanal multipoint full-layer puncture biopsy (TMFP) in predicting pathological complete response (pCR) after neoadjuvant radiotherapy and chemotherapy (nCRT) in patients with locally advanced rectal cancer (LARC) and to establish a predictive model for providing clinical guidance regarding the treatment of LARC.Methods:In this multicenter, prospective, cohort study, we collected data on 110 LARC patients from four hospitals between April 2020 and March 2023: Beijing Chaoyang Hospital of Capital Medical University (50 patients), Beijing Friendship Hospital of Capital Medical University (41 patients), Qilu Hospital of Shandong University (16 patients), and Zhongnan Hospital of Wuhan University (three patients). The patients had all received TMFP after completing standard nCRT. The variables studied included (1) clinicopathological characteristics; (2) clinical complete remission (cCR) and efficacy of TMFP in determining pCR after NCRT in LARC patients; and (3) hospital attended, sex, age, clinical T- and N-stages, distance between the lower margin of the tumor and the anal verge, baseline and post-radiotherapy serum carcinoembryonic antigen (CEA) and carbohydrate antigen (CA)19-9 concentrations, chemotherapy regimen, use of immunosuppressants with or without radiotherapy, radiation therapy dosage, interval between surgery and radiotherapy, surgical procedure, clinical T/N stage after radiotherapy, cCR, pathological results of TMFP, puncture method (endoscopic or percutaneous), and number and timing of punctures. Single-factor and multifactorial logistic regression analysis were used to determine the factors affecting pCR after NCRT in LARC patients. A prediction model was constructed based on the results of multivariat analysis and the performance of this model evaluated by analyzing subject work characteristics (ROC), calibration, and clinical decision-making (DCA) curves. pCR was defined as complete absence of tumor cells on microscopic examination of the surgical specimens of rectal cancer (including lymph node dissection) after NCRT, that is, ypT0+N0. cCR was defined according to the Chinese Neoadjuvant Rectal Cancer Waiting Watch Database Study Collaborative Group criteria after treatment, which specify an absence of ulceration and nodules on endoscopy; negative rectal palpation; no tumor signals on rectal MRI T2 and DWI sequences; normal serum CEA concentrations, and no evidence of recurrence on pelvic computed tomography/magnetic resonance imaging.Results:Of the 110 patients, 45 (40.9%) achieved pCR after nCRT, which was combined with immune checkpoint inhibitors in 34 (30.9%). cCR was diagnosed before puncture in 38 (34.5%) patients, 43 (39.1%) of the punctures being endoscopic. There were no complications of puncture such as enterocutaneous fistulae, vaginal injury, prostatic injury, or presacral bleeding . Only one (2.3%) patient had a small amount of blood in the stools, which was relieved by anal pressure. cCR had a sensitivity of 57.8% (26/45) for determining pCR, specificity of 81.5% (53/65), accuracy of 71.8% (79/110), positive predictive value 68.4% (26/38), and negative predictive value of 73.6% (53/72). In contrast, the sensitivity of TMFP pathology in determining pCR was 100% (45/45), specificity 66.2% (43/65), accuracy 80.0% (88/110), positive predictive value 67.2% (45/67), and negative predictive value 100.0% (43/43). In this study, the sensitivity of TMFP for pCR (100.0% vs. 57.8%, χ 2=24.09, P<0.001) was significantly higher than that for cCR. However, the accuracy of pCR did not differ significantly (80.0% vs. 71.8%, χ 2=2.01, P=0.156). Univariate and multivariate logistic regression analyses showed that a ≥4 cm distance between the lower edge of the tumor and the anal verge (OR=7.84, 95%CI: 1.48-41.45, P=0.015), non-cCR (OR=4.81, 95%CI: 1.39-16.69, P=0.013), and pathological diagnosis by TMFP (OR=114.29, the 95%CI: 11.07-1180.28, P<0.001) were risk factors for pCR after NCRT in LARC patients. Additionally, endoscopic puncture (OR=0.02, 95%CI: 0.05-0.77, P=0.020) was a protective factor for pCR after NCRT in LARC patients. The area under the ROC curve of the established prediction model was 0.934 (95%CI: 0.892-0.977), suggesting that the model has good discrimination. The calibration curve was relatively close to the ideal 45° reference line, indicating that the predicted values of the model were in good agreement with the actual values. A decision-making curve showed that the model had a good net clinical benefit. Conclusion:Our predictive model, which incorporates TMFP, has considerable accuracy in predicting pCR after nCRT in patients with locally advanced rectal cancer. This may provide a basis for more precisely selecting individualized therapy.