Objective To explore the the detection of MMP-2,-7,-9,and-12 enzymatic activity using the CEACAM1-derived fluorescent peptide substrate Site 84,investigating the application of substrate Site 84 to distinguishing between MMP-2 and MMP-9 in the gelatinase spectrum of MMPs.Methods The fluorescent enzymatic method was employed to observe the detection of MMP-2,-7,-9,and-12 enzymatic activity using substrate Site 84;further observations were made on the sensitivity and specificity of substrate Site 84 to enzymatic activity of MMP-2 and MMP-9 within the gelatinase spectrum;the kinetic parameters(Km and Kcat)of the enzymatic reaction between substrate Site 84 and MMP-2 were obtained.Results Using Site 84 as a substrate,enzymic kinetics curves for MMP-12,-7,-2 were obtained,but no enzymatic activity curve for MMP-9 was observed.Furthermore,Site 84 specifically detected the enzymatic activity of MMP-2 within the gelatinase spectrum,capable of detecting low concentration(0.6 μM)of MMP-2 enzymatic activity,but no obvious enzymatic reaction was observed for high concentration(6 μM)of MMP-9;the kinetics parameters for the enzymatic reaction between Site 84 and MMP-2 were Km=315 μM,Kcat/Km=2 565/MS.Conclusion The CEACAM1-derived substrate Site 84 serves as a novel fluorescent peptide substrate,enabling the acquisition of enzymatic activity curves for MMP-12,-7 and-2,and specifically detecting the enzymatic activity of MMP-2 within the MMP gelatinase spectrum.

ObjectiveTo observe the effects of Hedysari Radix polysaccharide on the apoptosis of gastric sinus smooth muscle cells and explore the underlying mechanism via the insulin-like growth factor-1 （IGF-1）/phosphatidylinositol 3-kinase （PI3K）/serine-threonine kinase （Akt） pathway in the rat model of diabetic gastroparesis （DGP）. MethodSixty-two Wistar male rats were randomized into a blank group （n=12） and a modelling group （n=50）. The rat model of DGP was established by small-dose multiple intraperitoneal injections of streptozotocin combined with an irregular high-fat and high-sugar diet for 4 weeks. The modeled rats were randomized into model group， mosapride citrate （1.35 mg·kg^-1）， and high-， medium-， and low-dose （200， 100， and 50 mg·kg^-1， respectively） Hedysari Radix polysaccharide groups. The rats were administrated with corresponding drugs by gavage， and those in the blank and model groups with equal volumes of pure water by gavage once a day for 8 consecutive weeks. The random blood glucose and body mass were measured every 2 weeks， and gastric emptying rate was calculated. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of smooth muscle in gastric antrum， and terminal deoxynucleoitidyl transferase-mediated nick-end labeling （TUNEL） was employed to detect the apoptosis of smooth muscle cells in the gastric antrum. The expression of IGF-1， phosphorylated （p）-PI3K， and p-Akt in the smooth muscle of gastric sinus tissue was detected by immunohistochemistry. Western blot was employed to determine the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in the smooth muscle of the gastric antrum. ResultCompared with the blank group， the model group showed elevated random blood glucose at all time points （P<0.01）， decreased body mass and gastric emptying rate （P<0.01）， increased apoptotic index of smooth muscle cells in the gastric antrum （P<0.01）， down-regulated protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and up-regulated protein level of Bax （P<0.01）. Compared with the model group， the 8 weeks of drug administration lowered the random blood glucose， increased the body mass and gastric emptying rate （P<0.05， P<0.01）， decreased the apoptotic index of smooth muscle cells in the gastric antrum （P<0.05， P<0.01）， up-regulated the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and down-regulated the protein level of Bax （P<0.05， P<0.01）. Compared with the mosapride citrate group，the administration of low-dose Hedysari Radix polysaccharide for 6 and 8 weeks lowered the random blood glucose and decreased the body mass （P<0.05， P<0.01），low and medium-dose Hedysari Radix polysaccharide decreased the gastric emptying rate and the apoptotic index of smooth muscle cells in the astragaloside low-dose group decreased （P<0.05）. The protein levels of IGF-1，p-PI3K/PI3K，p-Akt/Akt and Bcl-2（low dose）were down-regulated and the protein level of Bax was up-regulated by low doses of Hedysari Radix polysaccharide （P<0.05， P<0.01）. Compared with high-dose Hedysari Radix polysaccharide， low-dose Hedysari Radix polysaccharide elevated random blood glucose and reduced body mass after 6 and 8 weeks of administration （P<0.05， P<0.01）， and the low and medium doses decreased the gastric emptying rate， increased the apoptotic index of smooth muscle cells in the gastric antrum （P<0.05， P<0.01）， down-regulated the protein levels of IGF-1， p-PI3K/PI3K， p-Akt/Akt， and Bcl-2， and up-regulated the protein level of Bax （P<0.05， P<0.01）. Compared with the medium-dose group，the low-dose group of Hedysari Radix polysaccharide had lower body mass，lower gastric emptying rate in rats，higher apoptotic index of smooth muscle cells in gastric sinus tissue after 6 and 8 weeks of administration （P<0.05， P<0.01）， and lower protein expression of IGF-1，p-PI3K/PI3K，p-Akt/Akt. ConclusionHedysari Radix polysaccharide protects the smooth muscle cells in gastric antrum against apoptotic injury and promotes gastric motility by activating the IGF-1/PI3K/Akt signaling pathway， as manifested by the up-regulated expression of IGF-1， p-PI3K， p-Akt， and Bcl-2 and down-regulated expression of Bax.

Objective:To discuss the effect of N-myc downstream-regulated gene 1(NDRG1)on the enzalutamide(ENZA)resistance in the castration-resistant prostate cancer(CRPC),and to clarify its mechanism.Methods:The human CRPC C4-2 cells and ENZA-resistant strain C4-2/ENZA cells were cultured in vitro.The expression levels of NDRG1 mRNA in the C4-2/ENZA cells and their parental C4-2 cells were detected by real-time fluorescence quantitative PCR(RT-qPCR)method.The expression levels of NDRG1,androgen receptor(AR),and prostate-specific antigen(PSA)proteins in the cells were detected by Western blotting method to verify the transfection efficiency of the cells.The C4-2/ENZA cells were divided into blank group(normally cultured without treatment),negative control lentivirus(Lv-NC)group(transfected with Lv-NC),Lv-NDRG1 group(transfected with Lv-NDRG1),Lv-NC+ENZA group(transfected with Lv-NC followed by ENZA treatment),Lv-NDRG1+ENZA group(transfected with Lv-NDRG1 followed by ENZA treatment),Lv-NDRG1+epidermal growth factor(EGF)group(transfected with Lv-NDRG1 followed by EGF treatment),and Lv-NDRG1+EGF+ENZA group(transfected with Lv-NDRG1 followed by EGF and ENZA treatment).The half-maximal inhibitory concentration(IC50),resistance index(RI),and proliferation activity of the cells were detected by MTT assay;the apoptotic rates of the cells in various groups were detected by flow cytometry;RT-qPCR method was used to detect the expression levels of NDRG1 mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of NDRG1,AR,phosphorylated AR at serine213(p-ARSer213),phosphorylated AR at serine81(p-ARSer81),and PSA proteins in the cells in various groups.Results:Compared with C4-2 cells,the expression levels of NDRG1 mRNA and protein in the C4-2/ENZA cells were significantly decreased(P<0.01)and the expression levels of AR and PSA proteins were increased(P<0.01),indicating low expression of NDRG1 in the ENZA-resistant C4-2/ENZA strain.Compared with Lv-NC group,the expression levels of NDRG1 mRNA and protein in the cells in Lv-NDRG1 group were significantly increased(P<0.01),indicating the successful construction of an NDRG1 gene over-expression strain of C4-2/ENZA resistant cells.The MTT assay results showed that compared with the C4-2 cells,the IC50 of the C4-2/ENZA cells was increased(P<0.01)and the RI was 17.78;compared with Lv-NC group,the IC50 of the C4-2/ENZA cells in Lv-NDRG1 group was decreased(P<0.01).After 24 h of EGF treatment,compared with Lv-NC group,the IC50 of the C4-2/ENZA cells in Lv-NC+EGF group was significantly increased(P<0.01);compared with Lv-NDRG1 group,the IC50 of the C4-2/ENZA cells in Lv-NDRG1+EGF group was increased(P<0.01).Compared with before ENZA treatment,after 24 h of ENZA treatment,the proliferation activities of C4-2 and C4-2/ENZA cells were gradually decreased(F=223.80,P<0.01;F=81.46,P<0.01).Compared with Lv-NC group,the proliferation activity in the C4-2/ENZA cells in Lv-NDRG1 group after 24 h of ENZA treatment was significantly decreased(P<0.01).After 24 h of EGF treatment,compared with Lv-NC group,the proliferation activity of the C4-2/ENZA cells in Lv-NC+EGF group was significantly increased(P<0.01),while the the proliferation activity of the C4-2/ENZA cells in Lv-NDRG1+EGF group was significantly decreased(P<0.01).The chosen concentration and treatment duration for further testing were 10 000 μmol·L-1 ENZA and the intervention time was 24 h.The flow cytometry results showed that after 24 h of ENZA treatment,compared with Lv-NC group,the apoptotic rate of the cells in Lv-NDRG1 group was significantly increased(P<0.01);compared with Lv-NC+ENZA group,the apoptotic rate of the cells in Lv-NDRG1+ENZA group was significantly increased(P<0.01).After 24 h of EGF treatment,compared with Lv-NDRG1 group,the apoptotic rate of the cells in Lv-NDRG1+EGF group was significantly decreased(P<0.01),while the apoptotic rate of the cells in Lv-NDRG1+ENZA group was significantly increased(P<0.01);compared with Lv-NDRG1+ENZA group,the apoptotic rate of the cells in Lv-NDRG1+EGF+ENZA group was significantly decreased(P<0.01).The Western blotting results showed that after 24 h of ENZA treatment,compared with Lv-NC group,the expression levels of AR and PSA proteins and the ratio of p-ARSer213/AR and p-ARSer81/AR in the cells in Lv-NDRG1 group were significantly decreased(P<0.05 or P<0.01).After 24 h of EGF treatment,compared with Lv-NC group,the expression levels of AR and PSA proteins and the ratio of p-ARSer213/AR and p-ARSer81/AR in the cells in Lv-NC+EGF group were significantly increased(P<0.05 or P<0.01);compared with Lv-NDRG1 group,the expression levels of AR and PSA proteins and the ratio of p-ARSer213/AR and p-ARSer81/AR in the cells in Lv-NDRG1+EGF group were significantly increased(P<0.01).Conclusion:Over-expression of NDRG1 can reduce the resistance of CRPC to ENZA,and its mechanism may be related to the inhibition of AR signaling.

ObjectiveTo explore the clinical efficacy and safety of Qifu Yixin prescription in treating chronic heart failure in the patients with the syndrome of heart Qi deficiency， so as to provide clinical evidence for the treatment of chronic heart failure with this prescription and promote the clinical application and transformation of this prescription. MethodA total of 106 chronic heart failure patients with the syndrome of heart Qi deficiency who met the criteria in Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine （TCM） from September 2022 to May 2023 were selected and randomized into an observation group （53 cases） and a control group （53 cases）. Both groups received routine Western medicine treatment. In addition， the observation group received Qifu Yixin prescription， while the control group received placebo. The treatment course for both groups was 12 weeks. The New York heart association （NYHA） cardiac function grading， N-terminal pro-B-type natriuretic peptide （NT-proBNP）， soluble growth-stimulating expression gene 2 （sST2）， left ventricular ejection fraction （LVEF）， the ratio of early diastolic maximum mitral flow velocity （E） to early diastolic mitral annular motion velocity （e'） （E/e'）， left ventricular end diastolic diameter （LVEDD）， TCM syndrome scores， 6-minute walking test （6MWT）， Minnesota Living with Heart Failure Questionnaire （MLHFQ）， and Kansas City Cardiomyopathy Questionnaire （KCCQ） were determined before and after treatment. ResultFinally， 102 patients were included for analysis， including 51 patients in the observation group and 51 patients in the control group. After treatment， 42 patients in the observation group showed improved cardiac function grading， with a total response rate of 82.35%， and 32 patients in the control group showed improved cardiac function grading， with a total response rate of 62.75%. The total response rate regarding the cardiac function in the observation group was higher than that in the control group （χ²=4.923， P<0.05）. The observation group outperformed the control group in lowering the NT-proBNP level， elevating LVEF， decreasing the E/e' ratio （P<0.05）， reducing LVEDD and sST2 levels， and recovering TCM syndrome score， 6MWT score， MLHFQ score， and KCCQ score （P<0.05）. None of the safety indexes in the two groups showed abnormal values before and after treatment， and no serious adverse reaction was observed. ConclusionQifu Yixin prescription can improve the heart function， exercise tolerance， and quality of life and alleviate the TCM syndrome of the chronic heart failure patients with the syndrome of heart Qi deficiency by inhibiting myocardial fibrosis.

Objective:To investigate the types, incidences, and clinical characteristics of shock in polytrauma patients at different stages after polytrauma.Methods:A retrospective study was conducted on polytrauma patients admitted to multiple trauma centers from June 2020 to December 2021. The inclusion criteria were patients >18 years old and treated due to polytrauma. Exclusion criteria included an admission time of more than 48 h after trauma, a history of malignancy, or metabolic, consumptive, and immunological diseases. The early stage was defined as the period of ≤48 h after polytrauma, and the middle stage was defined as the period between 48 h and 14 days. The patient’s medical history, clinical manifestations, laboratory tests, imaging examination, injury severity score (ISS), and Glasgow coma scale (GCS) were collected. The types, incidences, and clinical characteristics of shock in different stages after polytrauma were analyzed, according to the diagnostic criteria of each type of shock. The differences between the groups were compared by Student’s t test, χ2 test or Mann-Whitney U test. Results:The incidence of the early and middle stage shock after polytrauma were 73.1% and 36.4%, respectively, with statistically significant difference between stages ( P<0.01). There were significant differences in the incidence of hypovolemic shock (83.6% vs. 28.4%), distributed shock (13.7% vs. 80.9%) and cardiogenic shock (3.5% vs. 6.6%) between stages (all P<0.05). The incidence of obstructive shock (8.4% vs. 9.7%, P>0.05) was similar between stages. The incidence of undifferentiated shock was 1.6% and 1.2%, respectively. There were 9.5% patients with multifactorial shock in the early stage and 14.4% in the middle stage. Totally 7 combinations of multifactorial shock were found in different stages after polytrauma. In the early stage, the combination of HS and DS accounted the highest ratio (42.3%) and followed by HS and OS for 28.8%. In the middle stage, the combination of HS and DS was the most common (48.6%) and followed by DS and OS (24.3%). Conclusions:The incidence of shock in polytrauma patients is high. Different types of shock can occur simultaneously or sequentially. Therefore a comprehensive resuscitation strategy is significant to improve the success rate of treatment.

Objective:To investigate the changes of skeletal muscle-derived microparticles (SKMPs) and their scaving proteins lactadherin in peripheral blood of patients with perihip fractures and their effects on blood coagulation. It provides a theoretical basis for clinical evaluation of hypercoagulability in patients with perioperative hip fractures.Methods:A total of 44 patients (72.87±13.9 years old) with perihip fractures from October 2020 to January 2021 were included in the fracture group, and 40 healthy people were included in the group without fracture. Activated clotting time (ACT) in fracture group and control group was measured by coagulation and platelet function analyzer. The level of SKMPs in peripheral venous blood was determined by flow cytometry. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of MPs scaving proteins lactadherin in peripheral blood of fracture group and control group.Results:A total of 44 patients with perihip fractures were included. There were 33 (75%) femoral neck fractures and 11 (25%) intertrochanteric fractures. SKMPs concentration in peripheral blood of perihip fracture group 175.73/μl; postoperative fracture group 124.28/μl; Normal control group 90.69/μl. ACT was (377.5±76.2) s in the perihip fracture group, (498.1±97.1) s in the post-fracture group, and (505.7±44.0) s in the control group. Correlation analysis showed that there was a linear relationship between SKMPs count and ACT value in fracture patients ( r=-0.52, P<0.001). There was no significant correlation between SKMPs concentration and ACT value in peripheral venous blood of the post-fracture group ( P>0.05); Lactadherin protein concentration in perihip fracture group: 2 779.61 pg/ml; Fracture postoperative group 1 985.38 pg/ml; Normal control group 1 651.19 pg/ml. There is a positive correlation between Lactadherin concentration and SKMPs concentration with a correlation coefficient of 0.70. Conclusion:SKMPs plays a role in the hypercoagulability of patients with perihip fractures. Decompensation of lactadherin protein may be one of the causes of perioperative hypercoagulation in patients with hip fractures.

Abstract In order to achieve the goal of reducing the smoking rate among Chinese adults at ages of 15 years and above to below 20% by 2030, smoking cessation services require to be reinforced. The current Chinese smoking cessation service system includes short-term smoking cessation interventions and smoking cessation hotlines at the population level, and smoking cessation clinics at the individual level; however, these smoking cessation services suffer from problems of low awareness, low accessibility and low utilization rate. Based on the publications pertaining to smoking cessation services and released policy documents in China until June 2022, this review analyzes the current status, problems and causes of smoking cessation services, and proposes suggestions for improving the smoking cessation service system construction in China.

Objective:To discuss how to avoid the occurrence of adverse events and provides basis for improving the extracorporeal membrane oxygenation (ECMO) transport safety management to formulate the corresponding preventive measures through analyzing the causes and characteristics of adverse events in transport of ECMO.Methods:By using a self-designed ECMO transport observation table to collect data, with a retrospective study of adverse events in patients with ECMO transport in ECMO center of Beijing Chaoyang Hospital from January 2013 to June 2017, carrying out classification and analysis according to the causes of adverse events and the potential risks of the patients, thus put forward the feasible preventive measures.Results:There were 53 cases of ECMO transport in study period, with 18 cases (33.96%) of adverse events, among which the incidence of adverse events in inner-hospital transport was 34.21% (13/38) and that in inter-hospital transport was 33.33% (5/15). There was no patient died in ECMO transport. In the adverse events of ECMO transport, the main causes were related to transport staff, transport equipment and patient, which accounting for 1/3 of each. Among them, the most prominent was 4 cases (22.22%) of equipment lacking and 3 cases of battery and power supply (16.67%). In classification according to the risk degree of patients, 6 cases (33.33%) of third grade risk were found.Conclusions:It is safe and feasible to carry out ECMO transport in inner-hospital transport and inter-hospital transport based on ECMO transport team and transport process of this hospital. However the unexpected events of high risk or crisis of life is inevitable in ECMO transport. Through standardized training for ECMO team, with full assessment before transport, by the use of ECMO checklist and strict implementation of various transport processes and specifications, the incidence of adverse events in ECMO transport may be reduced.

Objective:To investigate the clinical characteristics of patients complicated with Hashimoto’s thyroiditis (HT) and papillary thyroid micro-carcinoma (PTMC) and risk factors for central lymph node metastasis (CLNM) by analyzing the clinical data.Methods:Clinical data of 770 patients with PTMC admitted to Head and Neck Surgery Center of Sichuan Cancer Hospital from May. 2015 to Nov. 2017 were retrospectively analyzed, including 250 HT-PTMC patients (observation group) and 520 non-HT-PTMC patients (control group) . There were 197 males and 573 females, with a male to female ratio of 1.00:2.91. Into observation indexes included patient’s age, gender, serum thyroid stimulating hormone (TSH) , thyroglobulin antibody (TG-Ab) , thyroid peroxidase antibody (TPO-Ab) , number of foci, diameter of foci, calcification of foci, location of foci (with or without extra-glandular invasion) , number of nodules (no matter benign or malignant) , and lymph node metastasis in central and lateral cervical regions. SPSS 22.0 software was used for statistical analysis. Normally distributed data were expressed ± s. The difference between observation group and control group was compared by chi-square test of single factor analysis. The risk factors of CLNM of the observation group were analyzed with multivariate Logistic regression, the difference was statistically significant if P<0.05. Results:There were statistically significant differences between the observation group and the control group in age ( P=0.006) , gender ( P<0.001) , TSH ( P<0.001) , TG-Ab ( P<0.001) , TPO-Ab ( P<0.001) , number of nodules ( P=0.016) , and central lymph node ( P<0.001) . Compared with non-HT-PTMC group, HT-PTMC group had a higher proportion of women under 55 years old, and both TG-Ab and TPO-Ab had higher positive rates. Patients with HT-PTMC were more likely to show polynodule changes, but their central lymph node metastasis rate was lower than that of the non-HT-PTMC group. Single-factor analysis showed that the number of cancer foci, calcification of cancer foci, and location of cancer foci were significantly correlated with the CLNM of HT-PTMC patients (all P<0.001) . Multivariate Logistic regression analysis showed that multiple cancer foci ( P<0.001) , invasion and capsule ( P<0.001) , and cancer foci with calcification ( P=0.005) were independent risk factors for CLNM. Conclusions:HT-PTMC is more common in women under 55 years of age, and most of them show multiple nodules in bilateral glandular lobes, often accompanied by elevated serum TSH, TG-Ab and TPO-Ab levels. Meanwhile, the lymph node metastasis rate of HT-PTMC is relatively low, and the prognosis may be relatively good. In HT-PTMC, the risk of multiple carcinoma foci, cancer foci with calcification, cancer foci invasion and the occurrence of CLNM is higher. Preventive central lymph node dissection helps to reduce the occurrence of postoperative cervical lymph node recurrence.

Objective: To explore the advantages and clinical efficacy of free chimeric perforator flap based on the descending branch of circumflex femoral artery applied to tongue reconstruction after advanced tongue cancer resection. Methods: From October 2013 to December 2018, 57 cases received tongue and oral base reconstruction surgeries using the descending branch of circumflex femoral artery chimeric perforator flap, including 39 males and 18 females, ranged from 20 to 76 years old. And all cases were with stage T3 and T4 tongue cancers, including 35 cases of squamous cell carcinoma, 7 cases of low differentiation cancer, 5 cases of oncosarcoma, and 10 cases of adenoid cystic carcinoma. The tongue was reconstructed by using perforator flap and muscle flap to fill the dead space at the oral floor. The artery anastomoses with the superior thyroid artery or facial artery in the receiving area, and the vein anastomoses with the internal jugular vein in the receiving area. The shape, function and local complications of the reconstructed tongue were observed after operation. Results: Of 57 cases, only one case had partial necrosis of flap, while other 56 cases with chimeric perforator flap survived. Postoperative gastric tube and tracheal cannula were removed in all patients, no cases with oral fistula. All donor sites were sutured in one stage. Postoperative radiotherapy was performed in 41 of the patients. All patients were followed up for 3 to 60 months (average of 20.7 months), with satisfactory esthetic and functional results in reconstructed tongues. Only linear scars were left in the donor areas of the legs, and no lower limb dysfunction was observed. Conclusions The descending branch of circumflex femoral artery chimeric perforator flap can used for repairing simultaneously the defects of both tongue and oral base. It is helpful to avoid the occurrence of oral fistula and to provide the reconstructed tongue with a good function. It is a good choice to use the descending branch of circumflex femoral artery chimeric perforator flap for tongue reconstruction after resection of advanced tongue cancer resection.