Objective To study the influencing elements in x-Gal staining method and optimize the reactive conditions so that nonspecific background can be eliminated and grafted exogenous cells carrying LacZ gene can be discerned correctly. Methods C17 2 cells (carrying lzcZ gene) were injected into the right lateral ventricle both in the adult and newborn animals. After one week they were perfused using two methods, then the slices were stained at different pH and incubating time respectively. The X-gal positive cells in hippocampus were counted under light microscope. Results Background staining in this method has close correlation with the species and age of the host animals and it decreases when pH is higher or incubating time is shorter. The results are the best when pH 9 5 and the incubating time is 1*!h.Conclusion The reliability of X-gal staining method depends on optimization of several parameters, including pH, incubating time, perfusion etc. It is necessary to establish the correspondent controls.;

Aim and Methods To investigate the effect of UCN-01(7-hydroxystaurosporine) on cell migration and invasion ability of DU-145, an invasive human prostate cancer cell line.Results It was found that UCN-01 at non-cytotoxic doses (100 nmol· L-1) significantly inhibited prostate cancer DU-145 cell invasion and migration behaviors.Moreover, this anti-invasion and migration activity of UCN-01 was associated with an up-regulation of cell adhesion molecule E-cadherin. Conclusion These results indicate for first time that UCN-01 inhibits the invasion and migration of human prostate cancer cells.Thus, clinical application of UCN-01 may contribute to the potential benefit for suppression of prostate cancer invasion and metastasis.

Objective To study the clinical presentations,radiologic features,prognosis,and possible causes of liver injury associated umbilical venous catheter (UVC ).Methods We reviewed database of our NICU from December 2012 to November 2015 and identified preterm infants with liver injury while UVC in place.The gestational age,birth weight,gender,days of UVC in place prior to liver injury, the depth of UVC (cm),UVC tip position,ultrasound findings of liver injury,laboratory tests,treatment and outcomes were collected.Results During study period,322 infants received UVC insertion.Ten cases (3.1 %)of liver injury associated with UVC were diagnosed.Of the ten infants,nine were diagnosed as extravasation of fluid to liver parenchyma,and one was diagnosed as liver hematoma.The mean birth weight was (1184 ±207)g,mean gestational age was (28.5 ±1.7)weeks.The UVC tip was at thoracic (T) vertebrae T 9 to T 11 level,nine UVC tips were below the right diaphragm level.All UVCs were used for total parenteral nutrition before liver injury was diagnosed.Six infants had clinical manifestations,and four infants were free of clinical manifestations.The most prominent clinical signs were abdominal distension (n =5),weak bowel sounds (n =5)and hepatomegaly (n =4).One infant who was diagnosed with liver hematoma also had progressive hemoglobin decline (minimum 34 g/L)and shock.Abdominal ultrasound showed well-limited,irregular,hyperechoic rimmed lesions with heterogeneously hypoechoic centers or anechoic liquid dark space.Extravasation of fluid to liver parenchyma will restorate within one week after the UVC was remove.One baby who was diagnosed as liver hematoma passed away at 9 days of life.Liver ultrasound of eight returned to normal in 52 days to 3.5 months,and one had calcified lesions at 9 months of age.Conclusions Liver injury is an unusual complication of UVC insertion and usage.Proper positioning of the UVC tip may help to avoid this complication.Early recognition,prompt diagnosis with liver ultrasound examination and timely treatment can lead to better outcome in newborns with extravasation of fluid to liver parenchyma.

Objective:To investigate the clinicopathological features of gastric adenocarcinoma of fundic gland type (GA-FG).Methods:A total of 12 patients, including 7 cases treated with endoscopic submucosal dissection (ESD), were diagnosed as having GA-FG in Nanjing Drum Tower Hospital from January 2018 to August 2019. Morphological changes were analyzed by reviewing endoscopic and pathological results. Patients were followed up after definitive diagnosis.Results:The clinical symptoms of patients with GA-FG were nonspecific. No Helicobacter pylori infection was identified. The lesions were found in the non-atrophic gastric mucosa of the upper 1/3 portion in 10 cases and middle 1/3 portion in 2 cases. Endoscopically, the most common features were whitish color (9 cases), and all lesions diameter≤1 cm. Their macroscopic types were classified as 0-Ⅰ (2 cases), 0-Ⅱa (9 cases) and 0-Ⅱc (1 case) respectively. All lesions had sharp boundary, with branching dilated blood vessels on the surface. Five in 7 cases who were treated with ESD showed submucosal invasion. Immunohistochemically, 9 cases were classified as the chief cell type , 3 as the mixed type, 11 MUC6 positive, 4 MUC5AC positive, 2 MUC2 positive, and 3 CD10 positive. P53 was detected in all 12 cases, and 9 cases had low Ki-67 staining index (<10%). The mean time of follow-up was 11 months, and 11 patients survived. Conclusion:GA-FG should be taken into consideration when the polyps are found in the upper part of the stomach, with whitish color, and branch dilated blood vessels on the surface. Excellent clinical outcomes can be achieved for GA-FG patients with ESD.

Objective:To evaluate the correlation of active inflammatory changes of the symphysis pubis on magnetic resonance imaging (MRI) scans with clinical factors in patients with axial spondyloarthritis.Methods:We retrospectively evaluated 112 patients with axial spondyloarthritis (ax-SpA) in our hospital from February 2014 to November 2020. Patients were divided into 4 groups: symphysis pubis + sacroiliac arthritis, symphysis pubis + non-sacroiliac arthritis, non-symphysis pubis + sacroiliac arthritis, and non symphysis pubis + non-sacroiliac arthritis group. Intra-group correlation coefficient ( ICC) was used to analyze the correlation between MRI active inflammation of the symphysis pubis and the sacroilioarthritis. Age, sex, symptom duration, smoking, body mass index, human leukocyte antigen (HLA)-B27 positive rate, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) among the four groups were compared by one-way analysis of variance (ANOVA) and Chi-square test. Results:The proportion of active inflammation of the symphysis pubis was 35.7% (40/112). There was no correlation between active inflammation of the symphysis pubis and sacroilioarthritis ( r=-0.06, P=0.559). Twenty-four patients of the 69 patients with sacroilioarthritis had active inflammation of the symphysis pubis, 16 patients of the 43 patients without sacroilioarthritis had active inflammation of the symphysis pubis. In patients without active inflammation of the sacroiliac joint, the CRP and ESR of the active inflammation of the symphysis pubis group was (49±60) mg/L, (40±19) mm/1 h, statistically higher than that of the non-active inflammation group (19±22) mg/L, (22±37) mm/1 h ( t=2.36, P=0.023; t=2.88, P=0.006). In patients who had active inflammation of the symphysis pubis, the symptom duration of the non-active inflammation of the sacroiliac joint, was (14±9) years, which was significantly longer than that of the active inflammation group (5±4) years ( t=4.07, P=0.001). Conclusion:There is no correlation between active inflammatory changes of the symphysis pubis and bone marrow edema of the sacroiliac joint. Therefore, in ax-SpA patients with inflammatory low back pain and/or hip/groin pain, and also with high levels of CRP, ESR, but no active inflammatory changes of the sacroiliac joint on MRI scans, active inflammation of the symphysis pubis should be considered.

Objective In patients with non -small cell lung cancer(NSCLC)undergoing radical surgery,there were still many inevitable recurrences and distant metastases,even after systemic postoperative adjuvant chemotherapy.At the same time,many patients were in the stage Ⅳ at the time of initial treatment.The aims of this study were to investigate and compare the first-line chemotherapy(First-line Chemotherapy at Recurrence Post-adjuvant Chemotherapy,FCRPC)in NSCLC patients with distant metas-tasis after adjuvant chemotherapy with initial treatment at the phase Ⅳ of NSCLC patients with first-line chemotherapy(Initial First-line Chemotherapy,IFC).Methods A total of 603 patients with distant metastatic NSCLC were collected in this study.Among them,73 of them were FCRPC and 530 of them for IFC.Statistical methods for propensity score matching were used to balance the clinical features between FCRPC and IFC groups.Chi-square test was used to compare the short-term efficacy between FCRPC and IFC groups.Survival analysis was performed using regression analysis and Kaplan-Meier analysis.Results There was no significant difference in objective response rate(ORR)and disease control rate(DCR)between FCRPC and IFC groups in NSCLC patients with dis-tant metastases(ORR rate:27.46% in the FCRPC group,24.7% in the PFC group,P=0.851 and DCR rate:78.1% in the FCRPC group,65.6% in the PFC group,P=0.140).There was also no significant difference in the median progression-free survival(9.8 months in the FCRPC group and 8.5 months in the PFC group,P=0.337)and median overall survival(20.0 months in the FCRPC group and 14.4 months in the PFC group,P=0.087).Conclusion There is no significant difference in the prognosis of first-line chemotherapy between NSCLC patients with distant metastases and with initial treatment at the stageⅣafter adjuvant chemotherapy.

Objective To assess the safety and feasibility of video electroencephalographic (VEEG) monitoring in preterm infants and critically ill neonates in neonatal intensive care unit (NICU). Method From December 2017 to June 2018, high risk infants were prospectively enrolled and received VEEG monitoring in our NICU. Their basic information, adverse events and disturbances of any procedure during VEEG monitoring were collected by specially-trained nurses. Result A total of 245 times of VEEG were recorded. The average gestational age (GA) was (32.1 ± 3.6) weeks, the birth weight (BW) was (1879 ± 757) g, the corrected GA (cGA) at VEEG monitoring was (33.8±3.3) weeks, and the average weight at VEEG monitoring was (2008±716) g. The earliest cGA at VEEG monitoring was 25+5 weeks, and the lowest weight at VEEG monitoring was 520 g. The average monitoring duration was (4.9±2.4) h, ranging from 3 to 20 hours. During VEEG monitoring, 80 cases (32.7％) received noninvasive ventilation, 43 cases (17.6％) mechanical ventilation. The adverse events during electrodes placement were oxygen desaturation in 8(3.3％) cases. During VEEG monitoring, local skin erythema were found in 4 cases (1.6％), and electrodes displacement in 2(0.8％) cases. The disturbances of any clinical procedures were reported in 18(7.3％) cases. No severe adverse events such as displacement of endotracheal tube nor events requiring cardiopulmonary resuscitation occurred during VEEG monitoring. Conclusion It is feasible and safe for trained NICU nurses to place electrodes for high risk neonates.

BACKGROUND:In the initial stage of multiple sclerosis,central immune cells activate and release a large number of inflammatory factors,causing white matter demyelination and even involving gray matter neurons.The equilibrium of differentiation between different subsets of CD4+ T cells plays an important role in the progression of experimental autoimmune encephalomyelitis.The previous results of the research group showed that the active ingredient astragalus glycoprotein in astragalus can regulate the immune response in experimental autoimmune encephalomyelitis mice,and whether it has a regulatory effect on the differentiation of T cell subsets has not been determined. OBJECTIVE:To explore the therapeutic effects and immune regulatory mechanisms of astragaloside IV on experimental autoimmune encephalomyelitis mice. METHODS:Female C57BL/6 mice were divided into the normal control group,experimental autoimmune encephalomyelitis disease model group,and astragaloside IV treatment group(n=8 per group).Myelin oligodendrocyte glycoprotein peptides 35-55 were used for experimental autoimmune encephalomyelitis model induction in the last two groups.On day 10 to 28 after immunization,the astragaloside IV treatment group was treated with 40 mg/kg per day astragaloside IV intragastrically.Body weight and clinical scores of mice in each group were recorded from the immunization day to the 28th day.On the 28th day after immunization,the mouse spinal cord was taken and made into frozen sections for hematoxylin-eosin staining and Lux fast blue staining to observe pathological changes in the spinal cord.Percentage of splenic T cell subsets was detected using flow cytometry.Western blot assay was used to determine the protein expression of interferon-γ,interleukin-17 and interleukin-6 in the spinal cord.Levels of interferon-γ,interleukin-17,interleukin-6 and interleukin-4 in supernatants of cultured splenocytes were determined by ELISA. RESULTS AND CONCLUSION:(1)Compared with the experimental autoimmune encephalomyelitis disease model group,astragaloside IV could reduce the degree of weight loss in experimental autoimmune encephalomyelitis mice(P<0.05),ameliorate clinical symptoms(P<0.05),inhibit the infiltration of inflammatory cells and alleviate myelin loss(P<0.01,P<0.05).(2)Compared with the experimental autoimmune encephalomyelitis disease model group,astragaloside IV could inhibit the proportion of CD4+T cell subsets expressing interferon-γ(P<0.001)and interleukin-17(P<0.001),but increase percentages of CD4+ interleukin-10+(P<0.001)and CD4+ transforming growth factor-β+(P<0.01)T cell subsets.(3)Astragaloside IV could inhibit the expression of interferon-γ(P<0.05,P<0.01),interleukin-17(P<0.05,P<0.05),and interleukin-6(P<0.05,P<0.05)in the spinal cord and spleen,and up-regulate the expression of interleukin-4(P<0.01)in spleen.(4)These findings confirm that astragaloside IV alleviates clinical symptoms in experimental autoimmune encephalomyelitis mice,which may be related to regulating the splenic T cell subsets,therefore,inhibiting the infiltration of inflammatory cells into the center and reducing the demyelination.

Objective:To evaluate the long-term efficacy of endoscopic transluminal drainage(ETD) for acute pancreatitis complicated with walled-off necrosis (WON) or pancreatic pseudocyst (PPC).Methods:A total of 79 patients who were diagnosed as having WON or PPC by abdominal CT or ultrasound and treated with ETD in Nanjing Drum Tower Hospital were enrolled. Past medical records and follow-up by phone call after discharge were analyzed for long-term outcomes including endocrine and exocrine functions and long-term quality of life.Results:A total of 50 patients were enrolled, including 31 patients with infected WON/PPC and 19 patients with uninfected WON/PPC. Seventeen patients (54.84%) in the infected WON/PPC group and 11 patients (57.89%) in the uninfected WON/PPC group lost 5% or more of their weight. There were no significant differences in the proportion of cases of weight loss of 5% or more ( P=0.833), or the weight loss between the two groups (12.59±8.89 kg VS 10.91±2.47 kg, P=0.522). Only one patient in the infected WON/PPC group had chronic abdominal pain. There was no significant difference in the Izbicki score between the two groups (23.79±6.74 VS 22.03±3.21, P=0.295). None of the patients developed steatorrhea after discharge. Five patients (16.67%, 5/30) in the infected WON/PPC group and 6 patients (40.00%, 6/15) in the uninfected WON/PPC group developed endocrine insufficiency with no significant difference ( P=0.140). Greater risk of secondary diabetes resulted from higher low-density lipoprotein cholesterol ( HR=1.9, 95% CI: 1.0-3.4, P=0.044)and triglycerides ( HR=1.2, 95% CI: 1.0-1.3, P =0.029). Conclusion:ETD is safe and effective for WON and PPC. But there is possibility that patients develop secondary diabetes. Additionally, greater risk of secondary diabetes results from higher low-density lipoprotein cholesterol and triglycerides.

Objective:To investigate the clinical, endoscopic and pathologic characteristics of gastric hyperplastic polyps coexisting with gastric cancers.Methods:A retrospective study was performed involving 18 patients who were pathologically confirmed with gastric hyperplastic polyps coexisting with gastric cancers. The clinical features, endoscopic findings, pathological characteristics and treatment strategy were analyzed.Results:The age of 18 patients was 67. 2±7. 8 years (ranged 45-79), including 6 males and 12 females. The symptoms included abdominal pain, distention, and some patients were asymptomatic. Multiple polyps (13/18) were common. Single lesions were often located in the gastric corpus (7/18). Endoscopy showed various morphological changes. Pedunculated polyp was the most common type (15/18). All polyps were larger than 10 mm in diameter, and the polyps in 9 patients were larger than 20 mm. Fourteen patients had atrophic gastritis in the background mucosa, and 4 patients had Helicobacter pylori ( HP) infection. Conclusion:Gastric hyperplastic polyps coexisting with gastric cancers shows no specific symptoms. For HP (-) atrophic gastritis patients accompanied with multiple gastric polyps, malignant transformation of those larger and pedunculated polyps is of possibility.