Objective To explore the experiment condition and method for the application of in vitro in vasive Transwell chamber and to observe muscarinicreceptor stimulant and muscarinicreceptor antagonist's influence to cholangiocarcinoma's invasiveness.Methods Two hundred microliter cell suspension of various concentrations(0.5×105/mL,1.0×105/mL,1.5×105/mL and 2.0×105/mL)was added into the upper chamber of the Transwell chamber,and the cells were allowed to penetrate the matrigel for 12,18,24and 48 hours respectively.The numbers was gotten as the invasive cells on the under surface of the membrane.After optimal cell concentration and time were gotten,pilocarpine of various concentrations(0 mmol/L,0.1 mmol/L,0.3 mmol/L and 0.5 mmoL/L)was added into the upper chamber of the Transwell chamber,then the cells on the matrigel were stained and counted.So did the cells when atropine of various concentrations(0.01 mmol/L,0.01 mmol/L,0.05 mmoVL and 0.1 mmol/L)were added into the upper chamher of the Transwell chamber in according to pilocarpine of various concentrations(0 mmol/L,0.3 mmol/L,0.3 mmol/L and 0.3mmol/L).Results With the increase of the time and cell concentrations，the cells couts that penetrated the matrigel increased，while the increase tended to he stable when the culture time exceeded 36 hous and the cell concentration Was over 1.0×105/mL.By adding pilocarpine,there were significant differences between the control and experimental groups(P＜0.05)，but there were no significant differences in experimental groups with various concentrations.There were no significant differences in blank group and experimental groups with atropine added(P＞0.05).When added pilocarpine and atropine,there were significant differences between blank and experimental groups(P＜0.05)，but there were no significant differences in experimental groups with various concentrations.Conclusions Thirty-six hours as invasive time,and one cell concentration 1.0 × 105/mL were optimal to test invasion abilities of cholangiocarcingma cells to different medicines or reagents.There is the possibility that museariniereceptor exists in cholangiocarcinoma cells，and may play an important role in cholangiocarcinoma's invasiveness and metastasis.

BACKGROUND:Previous studies have confirmed that bone marrow mesenchymal stem cells in vitro can promote hepatic stel ate cellapoptosis and inhibit its activity, in which the mechanism of action remains unknown. OBJECTIVE:To screen out apoptosis-related genes during hepatic stel ate cellapoptosis regulated by bone marrow mesenchymal stem cells using gene chip technology. METHODS:Purified human bone marrow mesenchymal stem cells were seeded in 6-wel Transwel plate and cocultured with hepatic stel ate cells. Cultured human bone marrow mesenchymal stem cells alone served as control group, and cultured for 72 hours. The alterations in apoptosis-related genes were analyzed between culture alone group and coculture group using gene chip technology. The genes strongly associated with regulation of hepatic stel ate cells were selected. RESULTS AND CONCLUSION:By the functional classification of second-generation SABiosciences Gene chips, apoptotic gene screening found that after coculture, significantly upregulated genes in bone marrow mesenchymal stem cells contained:AKT1, PIK3R2, DAPK1, DHCR24, NOTCH2 and BDNF. Combined with previous findings, we hypothesized that NOTCH may play a key role in the regulation of hepatic stel ate cells by bone marrow mesenchymal stem cells.

Objective:To evaluate the changes in topological properties of brain functional network after induction of general anesthesia in the patients with glioma.Methods:Twenty-two patients scheduled for elective intracranial glioma resection were selected.Resting-state functional magnetic resonance imaging was performed during wakefulness and general anesthesia with endotracheal intubation in patients with glioma. Large-scale functional brain networks of each patient were constructed based on 123 regions of interest in non-surgical hemisphere. Global properties (local efficiency, clustering parameter, shortest path length, global efficiency, small world) and nodal properties (nodal degree, nodal efficiency, and between centrality) in brain functional networks were then compared between wakefulness and general anesthesia.Results:Eighteen patients were finally enrolled. Compared with the status during wakefulness, the local efficiency and clustering parameter on non-surgical side significantly decreased ( P<0.05), no significant change was found in the shortest path length and global efficiency ( P>0.05), and small world was greater than 1 throughout the entire density range; the nodal degree, nodal efficiency and between centrality of nodes located in the medial/mesal regions, such as the medial prefrontal cortex, posterior cingulate gyrus/precuneus, medial temporal lobe, anterior cingulate gyrus, thalamus, amygdala, were significantly reduced ( P<0.05); however, these node parameters increased significantly in the lateral brain regions ( P<0.05) except for the primary auditory and somatosensory cortex, which also decreased significantly after induction of general anesthesia( P<0.05). Conclusions:The functional segregation of brain functional network is widely inhibited after induction of general anesthesia, but the functional integration is still retained. The lateral brain regions show no anticorrelation with the medial brain region during general anesthesia.

ObjectiveTo investigate the inhibition of ENAH gene silenced by siRNA on the growth of liver cancer cell.MethodsENAH-siRNA and control-siRNA were transfected to human liver cancer cell HCCLM3 using liposome Lipofectamine 2000 and ENAH-siRNA group and control-siRNA group were established. The expression of ENAH protein was detected by Western blot. Tumorigenic ability of the liver cancer cell was observed through nude mice tumorigenicity assay. Tumor volume was recorded, growth curve was drawn and survival analysis was conducted. The experimental data of two groups were compared usingt test, and the survival analysis was conducted using Kaplan-Meier method and Log-rank test.ResultsThe expression of ENAH protein in ENAH-siRNA group was significantly less than that in control-siRNA group. The formation time of solid tumor after inoculation in nude mice in ENAH-siRNA group was (24±3) d, which was significantly longer than (8±2) d in control-siRNA group (t=12.55,P<0.05). The median survival time of the nude micewas 64 (48~81) d in ENAH-siRNA group and was 34 (21~48) d in control-siRNA group. There was significant difference in the overall survival rate between two groups (χ2=14.33,P<0.05).Conclusion ENAH gene silenced by siRNA may obviously weaken the tumorigenic ability of liver cancer cell and inhibit the growth of tumor.

ObjectiveTo investigate the clinical characteristics of primary liver cancer (PLC) complicated with non-Hodgkin’s lymphoma.MethodsClinical data of 2 patients with PLC complicated with non-Hodgkin's lymphoma admitted and treated in the Third Afifliated Hospital of Sun Yat-sen University between January 2006 and July 2015 as well as 18 patients reported by the literature were retrospectively analyzed. The informed consents of all patients were obtained and the local ethical committee approval was received. The incidence, diagnosis and treatment process, therapeutic regimen, curative effect and prognosis were observed. Relevant literature in PubMed database from January 1994 to December 2015 was searched for literature review.ResultsAmong the patients, 17 were males and 3 were females with the onset age ranging from 35 to 80 years old and the median of 64 years old. Nineteen cases were with hepatocellular carcinoma and 1 with mixed type liver cancer. Hepatitis virus infection was found in 90%(18/20) of the patients with 8 cases of hepatitis B virus (HBV) infection and 10 of hepatitis C virus (HCV) infection. All patients were complicated with B-cell non-Hodgkin's lymphoma. The main type was diffuse large B-cell lymphoma, accounting for 60%(12/20). And the other types were follicular lymphoma (n=5), mucosa-associated lymphoid tissue (MALT) lymphoma (n=1), marginal zone lymphoma (n=1) and unspeciifed lymphoma (n=1). The main lesions of non-Hodgkin's lymphoma respectively located in the liver (n=9), spleen (n=3), lymph node (n=3), stomach (n=3), vertebral body (n=1), other non-tissue and visceral organ (n=1). The therapeutic regimens were operation + chemotherapy (n=9), radiofrequency ablation or transcatheter arterial chemoembolization (TACE) + chemotherapy (n=6), liver transplantation (n=1) and palliative treatment (n=4). The median postoperative survival time of the patients was 48(5-105) months, while the survival time of the patients receiving palliative treatment was less than 1 month.ConclusionsMost PLC patients complicated with non-Hodgkin's lymphoma are male, and the pathological type of all the non-Hodgkin's lymphoma is B-cell type. The morbidity is closely associated with HBV infection. There are no speciifc clinical manifestations, and the conifrmed diagnosis depends on the pathological examination. Radical resection is the ifrst choice for treatment.

Objective:To identify the risk factors for delayed gastric emptying in patients scheduled for elective caesarean section.Methods:One hundred and five patients, aged≥18 yr, at≥37 week gestation, of American Society of Anesthesiologists Physical Status classification Ⅱ or Ⅲ, scheduled for elective caesarean section from August 2023 to February 2024, who followed the preoperative fasting guidelines, were collected. Based on the quantitative and qualitative ultrasound assessments of the gastric antrum, patients were divided into 2 groups: delayed gastric emptying group, as defined by Perlas A score of grade 2 or the presence of solid contents or gastric volume/weight >1.5 ml/kg, and non-delayed gastric emptying group, as defined by Perlas A score < grade 2 and gastric volume/weight ≤1.5 ml/kg. The variables of which showed statistically significant differences in the intergroup comparison would enter the logistic regression analysis to stratify the risk factors for delayed gastric emptying before elective caesarean section.Results:There were 41 parturients (39.0%) showed delayed gastric emptying before elective caesarean section. According to the results of multifactorial logistic regression analysis, the gastroesophageal reflux disease( OR=5.313, 95% confidence interval 1.013-27.878, P=0.048) was an independent risk factor for delayed gastric emptying in the patients who underwent elective caesarean section. Conclusions:Gastroesophageal reflux disease is an independent risk factor for delayed gastric emptying in the patients scheduled for elective caesarean section.

Objective:To study the efficiency and difference of the artificial intelligence (AI) system based on fundus-reading in community and hospital scenarios in screening/diagnosing diabetic retinopathy (DR) among aged population, and further evaluate its application value.Methods:A combination of retrospective and prospective study. The clinical data of 1 608 elderly patients with diabetes were continuously treated in Henan Eye Hospital & Henan Eye Institute from July 2018 to March 2021, were collected. Among them, there were 659 males and 949 females; median age was 64 years old. From December 2018 to April 2019, 496 elderly diabetes patients were prospectively recruited in the community. Among them, there were 202 males and 294 female; median age was 62 years old. An ophthalmologist or a trained endocrinologist performed a non-mydriatic fundus color photographic examination in both eyes, and a 45° frontal radiograph was taken with the central fovea as the central posterior pole. The AI system was developed based on the deep learning YOLO source code, AI system based on the deep learning algorithm was applied in final diagnosis reporting by the"AI+manual-check" method. The diagnosis of DR were classified into 0-4 stage. The 2-4 stage patients were classified into referral DR group.Results:A total of 1 989 cases (94.5%, 1 989/2 104) were read by AI, of which 437 (88.1%, 437/496) and 1 552 (96.5%, 1 552/1 608) from the community and hospital, respectively. The reading rate of AI films from community sources was lower than that from hospital sources, and the difference was statistically significant ( χ2=51.612, P<0.001). The main reasons for poor image quality in the community were small pupil (47.1%, 24/51), cataract (19.6%, 10/51), and cataract combined with small pupil (21.6%, 11/51). The total negative rate of DR was 62.4% (1 241/1 989); among them, the community and hospital sources were 84.2% and 56.3%, respectively, and the AI diagnosis negative rate of community source was higher than that of hospital, and the difference was statistically significant ( χ2=113.108, P<0.001). AI diagnosis required referral to DR 20.2% (401/1 989). Among them, community and hospital sources were 6.4% and 24.0%, respectively. The rate of referral for DR for AI diagnosis from community sources was lower than that of hospitals, and the difference was statistically significant ( χ2=65.655, P<0.001). There was a statistically significant difference in the composition ratio of patients with different stages of DR diagnosed by AI from different sources ( χ2=13.435, P=0.001). Among them, community-derived patients were mainly DR without referral (52.2%, 36/69); hospital-derived patients were mainly DR requiring referral (54.9%, 373/679), and the detection rate of treated DR was higher (14.3%). The first rank of the order of the fundus lesions number automatically identified by AI was drusen (68.4%) and intraretinal hemorrhage (48.5%) in the communities and hospitals respectively. Conclusions:It is more suitable for early and negative DR screening for its high non-referral DR detection rate in the community. Whilst referral DR were mainly found in hospital scenario.

Objective:To investigate the perioperative alterations and management of coagulation function in patients of massive blood transfusion during retroperitoneal tumor (RT)resection.Methods:Fourty-seven RT patients at Peking University International Hospital from Jan 2016 to Dec 2021 undergoing resection with massive blood transfusion more than 20 U within 24 h were reviewed for coagulation function before and after surgery.Results:Intraoperative bleeding was 3 000-25 800 ml, 10 patients had blood loss ≥10 000 ml. During the operation, (25.3±9.9) U of red blood cells were transfused, (2 720±1 369) ml plasma transfused, and (2.4±3.3) U platelets were transfused in 6 patients. Fourty-five patients received intraoperative albumin of (79.5±46.5) g; All 47 patients received fibrinogen of (2.3±1.3) g; Prothrombin complex was given in 45 patients (1 205±807) U. Preoperative hemoglobin was statistically different compared to postoperatively and days 1, 3 and 5 ( W=1 790, P<0.001; W=1 672, P<0.001; W=1 704, P<0.001; W=1 486, P=0.004);As with platelets, the difference was also statistically significant compared to postoperative days 1, 3, and 5 ( W=2 153, P<0.001; W=2 092, P<0.001; W=1 732, P<0.001); Preoperative albumin was different compared to postoperative days 1 and 3 ( W=1 568, P<0.001; W=1 578, P<0.001,); Preoperative fibrinogen was different compared to postoperative day 1 ( W=1 964, P<0.001). PT and APTT were prolonged on postoperative days 1 and 3 ( W=628, P<0.001, W=804, P=0.023) ( W=661, P<0.001, W=796, P=0.02). Patient's preoperative fibrin degradation products and D-dimer were above the normal value and were higher on postoperative days 3 and 5 ( W=498, P<0.001, W=345, P<0.001). Conclusions:Coagulation disorders occur perioperatively in patients with massive transfusion while undergoing surgery for RT.The implementation of ratiional transfusion strategy and close postoperative survey and management of coagulation dysfunction help avoid the coagulation related morbidities.

Objective To review the diagnosis and treatment of severe complications after transcatheter arterial chemoembolization (TACE) of hepatocellular carcinoma (HCC). Methods Clinical data of 15 patients with severe complications after TACE of HCC in Department of General Surgery, Lingnan Hospital, the Third Afifliated Hospital of Sun Yat-sen University from June 2011 to May 2013 were analyzed retrospectively. The informed consents of all patients were obtained and the ethics committee approval was received. There were 12 males and 3 females with age ranging from 40 to 78 years old and the median age of 55 years old. Results Manifestations of cholangitis such as abdominal pain, fever, rising white blood cell etc. were observed in all the patients. According to the clinical manifestations, history of receiving TACE and imaging examinations, 10 cases were diagnosed with biloma after TACE of HCC, 4 cases with liver abscess and 1 case with ischemic cholangitis. All the patients received basic treatments of anti-infection, cholagogue and liver protection. Ten cases with biloma were cured after receiving ultrasound-guided percutaneous transcatheter drainage of biloma, percutaneous transhepatic cholangial drainage (PTCD) and endoscopic nasobiliary drainage (ENBD). Four cases with liver abscess were cured after receiving ultrasound-guided percutaneous transcatheter drainage of liver abscess, local douche with antibiotics. One case with ischemic cholangitis received drainage of 3 catheters of PTCD and the serum bilirubin went down from 500μmol/L to 300μmol/L, but gave up treatment ifnally because of complicating severe biliary infection and gastrointestinal hemorrhage. Conclusions The diagnosis of severe complications after TACE of HCC is mainly based on the clinical manifestations, history of receiving TACE and imaging examinations. The therapies include basic treatments of anti-infection, cholagogue and liver protection etc., and symptomatic treatments of ultrasound-guided percutaneous drainage, PTCD, ENBD, etc.

Objective To investigate the impact of exogenous of triiodothyronine (T3) on the liver hyperplasia of mouse. Methods Forty-ifve healthy speciifc pathogen free (SPF) C57BL/6 mice were divided into group A, B and control group using random number table method with 15 mice in each group. Mice in group A, B were respectively injected with 2 ml exogenous T3 solutions 10, 5μg/kg intraperitoneally. Mice in control group were injected with 2 ml normal saline. Three mice of each group were put to death respectively on day 0, 7, 14, 21, 42 after treatment. The total liver weight of the mice was measured after death. The proliferation of liver cells was detected by immunohistochemistry. The experimental data were compared using t test or analysis of variance. Results Compared with control group, the liver weight of mice in group A increased signiifcantly on day 7, 14, 21, 42 after treatment (t=3.298, 6.760, 7.119, 6.128;P<0.05) , and the liver weight of mice in group B increased signiifcantly on day 14, 21, 42 after treatment (t=4.188, 4.570, 2.978;P<0.05). The increased liver weight in group A was signiifcantly more than that in group B on day 7, 14, 21, 42 after treatment (t=4.935, 4.303, 4.033, 4.480;P<0.05). The liver weight in group A, B rose to the top on day 21 after treatment (F=21.480, 11.244;P<0.05). Compared with control group, the liver cell count in group A increased signiifcantly on day 0, 7, 14, 21, 42 after treatment (t=28.383, 23.842, 40.194, 31.059, 15.841;P<0.05), and the same with group B (t=9.097, 7.680, 20.597, 42.192, 14.415;P<0.05). The increased liver cell count in group A was signiifcantly more than that in group B (t=8.016, 4.872, 10.719, 9.514, 7.831;P<0.05). The liver cell count rose to the top in group A on day 14 after treatment (F=169.190, P<0.05) and rose to the top in group B on day 21 after treatment (F=90.460, P<0.05). Extensive proliferation of liver cells was observed both in group A and B after treatment. Conclusions Exogenous T3 can effectively promotes the liver hyperplasia of mouse, and the hyperplasia becomes more signiifcant as the T3 concentration rises.