[Abstract ] Objective Difficulties with the preparation of gastric cancer stem cells (CSCs) have been a main obstacle to the studies of gastric cancer .This article addresses the technology of the serum-free medium suspension cultivation of the MKN-45 gastric cancer cell line and screening of stem cells from the cell line based on the biomarkers of gastric CSCs . Methods MKN-45 cells were cultured in serum-free medium for 8 weeks and those in the logarithmic phase cultivated with hoechst 33342 followed by detection of the side cells by flow cytometry .When the side population cells reached 25%, all the cell microspheres were collected , hatched with CD133 and CD44, and subjected to fluorescence-activated cell sorting.The CDl33 +and CD44 +cells were selected as gastric CSCs . Results About 40%of the MKN-45 gastric CSCs were alive , prolif-erated, and formed floating cell balls .Side population cells constitu-ted 3.4% of the MKN-45 cells and 26.9% of the cell balls.The CDl33 +and CD44 +cells made up 11.2% of the MKN-45 cells and 90.3%of the cell balls. Conclusion Cell balls rich in CSCs can be successfully obtained by serum-free medium suspension culti-vation and CSCs can be screened out with hoechst 33342 and surface markers , which may serve as an experimental ground for the stud-ies of gastric CSCs .

Objective To study whether osteoblast is necessary for IGF-Ⅰ to promote bone resorption by osteoclast.Methods Mouse MC3T3 osteoblast cells and mature osteoclasts induced by RANKL were cultured in vitro.These osteoblasts and osteoclasts were subjected to treatment with recombinant human insulin-like growth factor-1 (rhIGF-Ⅰ),and the activation of IGF-Ⅰ receptor was verified by Western blotting.Thereafter,osteoclasts were cultured individually or co-cultured with osteoblast,in the absence or presence of rhIGF-Ⅰ.Osteoclast proliferation and apoptosis were observed by MTT colorimetric assay and flow cytometry.Cathepsin K gene expression was detected by real-time PCR; bone adsorption activity of osteoclast was determined by resorption pits formation on calf cortex slice with toluidine blue staining.Results Western blotting result confirmed that rhIGF-Ⅰ could effectively activate IGF-Ⅰ receptors either in osteoblast or osteoclast.In co-cultured group,in the presence of rhIGF-Ⅰ osteoclast showed inhibited apoptosis,enhanced proliferation and up-regulated cathepsin K expression (P ＜ 0.05).The functional experiment revealed that osteoclasts collected from IGF-Ⅰ treated co-cultured group resulted in more resorption pits formation (P ＜ 0.05); rhIGF-Ⅰ did not show any significant effect on the individually cultured osteoclasts.Conclusion Osteoblast is necessary for osteoclast induced bone resorption resulting from IGF-Ⅰ treatment.

Objective:To observe the efficacy of apatinib combined with capecitabine in the treatment of advanced esophageal cancer.Methods:A total of 101 patients with advanced esophageal cancer in Taixing People's Hospital of Jiangsu Province from June 2017 to February 2018 were enrolled, and all the patients were divided into the control group (50 cases) and the observation group (51 cases) according to the random number table. The control group was treated with capecitabine combined with radiotherapy, and the observation group was treated with apatinib on the basis of the control group. The therapeutic effects, adverse reactions and progression-free survival (PFS) time of the two groups were compared.Results:The overall response rate in the observation group was higher than that in the control group [90.2% (46/51) vs. 72.0% (36/50)], and the difference was statistically significant ( χ2 = 5.473, P = 0.019). There were no significant differences in leukopenia, neutropenia, thrombocytopenia, anemia, proteinuria and hypertension between the two groups (all P > 0.05). The median PFS time in the observation group was 18.49 months (95% CI 15.35-25.03 months), and that in the control group was 13.33 months (95% CI 10.36-18.24 months), and the difference between the two groups was statistically significant ( χ2 = 5.995, P < 0.01). Conclusions:The therapeutic effect of apatinib combined with capecitabine in the treatment of advanced esophageal cancer is accurate. No obvious adverse reaction occurs, and the PFS time is prolonged.

Objective To investigate the effects of disulfiram combined with copper (DSF /Cu) on proliferation and migration of brain metastases from lung cancer. Methods The cell were divided into blank control group, valproate group (1.0 mmol·L-1 of valproate) , DSF /Cu (1,2,3,4,5 mg /0.17 mg) . Brain metastases from lung adenocarcinoma were incubated 72 hours.The changes of proliferative ability of cancer cells were detected by CCK-8 method, the effect of DSF /Cu on the migration ability of cancer cells was detected by Transwell chamber, and the degree of apoptosis of cancer cells was detected by Annexus V-PI double staining method. Results The ability of proliferation was significantly inhibited, the ability of migration was significantly reduced,and the rate of cell apoptosis was significantly increased in each dose group of brain metastases from lung adenocarcinoma compared with the blank control group. With the increase of DSF /Cu dose, its ability to inhibit the proliferation and migration of metastases cancer cells and induce apoptosis of metastases cancer cells has increased gradually. Conclusion DSF /Cu can inhibit the proliferation and migration of brain metastases from lung adenocarcinoma and induce apoptosis. DSF is expected to be a new method of the combined treatment of brain metastases cancer.

Objective To explore the relationship between different blood glucose levels and prognosis in patients with acute severe craniocerebral injury.Methods Clinical data of 120 patients with acute severe craniocerebral injury from Jun.2014 to Jun.2017 in Department of Neurosurgery,Tongren Hospital of Wuhan University were analyzed retrospectively.The patients were divided into the group of acute severe craniocerebral injury and the group of acute extra-severe craniocerebral injury according to the Glasgow coma score (GCS).Blood glucose levels were detected at the time of admission,the 1st day,the 3rd day,the 7th day and the 14th day after operation.According to the blood glucose levels on admission,the patients were divided into the normal blood glucose group,the blood glucose value of 6.1-10.0 mmol/L group,the blood glucose value ＞ 10.0 mmol/L group.The prognosis was evaluated by Glasgow Outcome Score (GOS) 3 months after discharge.The relationship between the blood glucose levels and GOS score in different groups was analyzed.Results The blood glucose levels in the group of acute extra-severe craniocerebral injury were higher than those in the group of acute severe craniocerebral injury,and the difference was statistically significant (P＜0.05).Of the patients with acute severe craniocerebral injury,54 had good prognosis,and 16 had poor prognosis.Of the patients with acute extra-severe craniocerebral injury,28 had good prognosis,and 22 had poor prognosis.The rate of good prognosis were 77.1％ and 56％ respectively,and the difference had statistical significance (P=0.014,x2=6.025).The good prognosis of patients with normal blood glucose group was 23 and the poor prognosis was 4,with a good prognosis rate of 85.2％ (23/27).Among patients with blood glucose of 6.1-10.0 mmol/L,30 had good prognosis and 8 had poor prognosis,with a good prognosis rate of 78.9％(30/38).Among patients with blood glucose ＞10.0 mmol/L,31 had good prognosis and 24 had poor prognosis,with a good prognosis rate of 56.4％ (31/55).With the increase of blood glucose,the good prognosis rate gradually decreased and the poor prognosis rate gradually increased,and the difference was statistically significant(P=0.010,x2=9.283).Conclusion Blood glucose level has influence on the prognosis of patients with acute severe craniocerebral injury,and the good prognosis of patients with acute severe craniocerebral injury is severely affected by hyperglycemia.

Objective: To investigate the clinicopathologic features of Rasmussen syndrome (RS) and to raise awareness of this rare disease. Methods: Clinicopathologic data of 4 cases of RS were retrospectively analyzed at Beijing Haidian Hospital from 2008 to 2016. Results: The clinical manifestations included epilepsia partialis continua and progressive neurologic deficits in all patients.MRI demonstrated unihemispheric focal cortical atrophy in all cases. The histopathologic changes included variable degrees of lymphocytic infiltrate within the cortex, subarachnoid space and perivascular cuffing.Microglial nodules and neuronophagia were seen. Mild to severe neuronal loss was noted with variable degrees of reactive gliosis. Spongy edema and cavitation were observed in focal cortex. Inflammation involving hippocampus was seen in one case. Three cases were accompanied by focal cortical dysplasia (FCD) Ⅲd. Immunohistochemical staining showed that the infiltrative lymphocytes were positive for CD3, CD8, granzyme B and TIA1 and the proliferating microglial cells were positive for CD68. NeuN positive neurons decreased significantly and reactive astrocytes were GFAP positive. Conclusions Pathologic changes of RS are similar to viral encephalitis and the inflammation is progressive and multifocal involving the hemisphere. The diagnosis of RS relies on pathologic features combined with clinical findings and neuroradiological examinations.

The present artificial anal sphincter fails to distinguish the different states of intestinal contents. In this study, we tried to distinguish the intestinal content by using a reflection type ultrasonic system, so as to lay the foundation for the development of artificial anal sphincter with sensing function. We measured the numbers of reflected waves when there were solid, liquid, gaseous or no contents in the recta of 30 Bama Miniature pigs. The results showed significant difference among the numbers of reflected waves of the 4 conditions ( = 1 088, < 0.05). Our research suggested the reflection-type ultrasonic system could be utilized to distinguish various contents inside the intestinal lumen, thus it might offer a new effective method for the development of artificial anal sphincter with sensing function.

Objective:To study the current status of longitudinal extrauterine growth restriction (EUGR) in extremely preterm infants (EPIs) and to develop a prediction model based on clinical data from multiple NICUs.Methods:From January 2017 to December 2018, EPIs admitted to 32 NICUs in North China were retrospectively studied. Their general conditions, nutritional support, complications during hospitalization and weight changes were reviewed. Weight loss between birth and discharge > 1SD was defined as longitudinal EUGR. The EPIs were assigned into longitudinal EUGR group and non-EUGR group and their nutritional support and weight changes were compared. The EPIs were randomly assigned into the training dataset and the validation dataset with a ratio of 7∶3. Univariate Cox regression analysis and multiple regression analysis were used in the training dataset to select the independent predictive factors. The best-fitting Nomogram model predicting longitudinal EUGR was established based on Akaike Information Criterion. The model was evaluated for discrimination efficacy, calibration and clinical decision curve analysis.Results:A total of 436 EPIs were included in this study, with a mean gestational age of (26.9±0.9) weeks and a birth weight of (989±171) g. The incidence of longitudinal EUGR was 82.3%(359/436). Seven variables (birth weight Z-score, weight loss, weight growth velocity, the proportion of breast milk ≥75% within 3 d before discharge, invasive mechanical ventilation ≥7 d, maternal antenatal corticosteroids use and bronchopulmonary dysplasia) were selected to establish the prediction model. The area under the receiver operating characteristic curve of the training dataset and the validation dataset were 0.870 (95% CI 0.820-0.920) and 0.879 (95% CI 0.815-0.942), suggesting good discrimination efficacy. The calibration curve indicated a good fit of the model ( P>0.05). The decision curve analysis showed positive net benefits at all thresholds. Conclusions:Currently, EPIs have a high incidence of longitudinal EUGR. The prediction model is helpful for early identification and intervention for EPIs with higher risks of longitudinal EUGR. It is necessary to expand the sample size and conduct prospective studies to optimize and validate the prediction model in the future.

Novel coronavirus pneumonia, which has emerged in Wuhan since the end of 2019, has posed a huge challenge for medical institutions in the city. Rapid completion of a number of cabin hospitals plays a vital role in preventing further spreading of the epidemic, by means of collecting and treating mild patients of the disease. This paper presents the key process of Tongji Hospital in its rapid informatization since it took over a cabin hospital. Based on the network architecture of the Tongji cloud platform, the shared service center is used to share data and integrate services between the cabin hospital and Tongji hospital. This practice can prevent cross-infection and improve service efficiency as well, hence offering a reference for future information infrastructure development of cabin hospitals.