BACKGROUND:Neural stem cells, as a hot topic in neuroscience research, have a wide application prospect in the treatment of neurological damage, but how to obtain a large number of terminally differentiated and purified nerve cells with homogeneous features is a difficult problem in this field. The use of intracellular fluorescence reporter system to track the process of neural stem cell differentiation and obtain a single kind of terminally differentiated and purified nerve cells provides a viable option. OBJECTIVE: To explore the value of GFAP promoter-driven fluorescence reporter system in tracing the neural differentiation of neural stem cells (NSCs). METHODS: Cerebral cortex of mouse embryos were primarily dissociated and sent for digesting and pipetting mechanically before suspension culture, followed by immunofluorescence staining of Nestin to identify their biological characteristics. Lentivirus carrying pLV/Final-neo-GFAP(promoter)-dTomato vector was employed to infect above-mentioned NSCs, and Geneticin (G418) was used to obtain purified NSCs at 14 days. Subsequently the purified cells were induced to differentiate into astrocyte-like cells; meanwhile red fluorescence changes in cells were observed by microscopy. The red fluorescent cells were then subjected to perform immunofluorescence staining at 13 days after induction. RESULTS AND CONCLUSION:The expression of Nestin in the isolated primary cells was strongly positive. Purified NSCs were obtained by lentivirus infection and subsequent G418 resistance selection at 14 days. After induced into astrocyte-like cells, the red fluorescence was observed in the cells under the microscope and furthermore, GFAP staining was also positive. Mouse NSCs carrying neo-GFAP(promoter)-dTomato were successfully obtained. The cells could express dTomato under the control of GFAP promoter, which provides a powerful tool for research on NSC differentiation mechanism, neural transplantation and tissue engineering product development.

Objective:To evaluate the efficacy of Epstein-Barr nuclear antigen 1/immunoglobulin A (EBNA1/IgA), BamH1 Z transactivator/IgA (Zta/IgA), capsid antigen/IgA (VCA/IgA), and Epstein-Barr virus deoxyribonucleic acid (EBV-DNA) in detecting different stages of na-sopharyngeal carcinoma (NPC). The relationship between the EBV markers and stages of NPC was also analyzed. Methods:Blood sam-ples of 152 untreated patients with NPC and 675 healthy subjects were collected.ELISA was used to detect the serum levels of EBNA1/IgA, Zta/IgA, and VCA/IgA. Fluorescence quantitative PCR (FQ-PCR) was used to detect the plasma levels of EBV-DNA. ROC and correla-tion analyses were employed to assess the detection assays for NPC diagnosis. The positive rates of EBV markers in NPC patients in dif-ferent stages were analyzed statistically. Results: The positive rates of EBNA1/IgA, Zta/IgA, VCA/IgA, and EBV-DNA in NPC patients were higher than those in the healthy individuals. The expression of EBNA1/IgA was relatively high in early NPC. The sensitivity of EB-NA1/IgA was 77.8%. In advanced NPC, the level of EBV-DNA was high, and the sensitivity of EBV-DNA was 88.8%. The specificity of EBV-DNA and EBNA1/IgA could reach more than 96%. The combination of EBV-DNA and EBNA1/IgA showed the best diagnostic value, with a sensitivity of 92.1%(early stage 82.5%, advanced stage 98.9%) and a specificity of 96.9%. The positive rates of EBV-DNA were positively associated with the NPC clinic stage and N stage. The positives rates of Zta/IgA were positively associated with the NPC N stage. Conclusion:The best single index for NPC screening in an asymptomatic population is EBNA1/IgA. EBV–DNA is an ideal index for auxiliary diagnostics of advanced NPC. The combination of EBV-DNA and EBNA1/IgA shows the best diagnostic value. EBV-DNA is an important index in the stage and illness monitoring of NPC. Zta/IgA can indirectly reflect the character of lymph node metastasis, and it may be useful in assessment of NPC surveillance.

Objective To study the effects of 3,5-diiodotyrosine (DIT) and potassium iodide (KI) on myocardial ATPase activity in hyperthyroidism Wistar rats induced by thyroid tablets.Methods Seventy-two Wistar rats were divided into 8 groups according to body weight by the random number table method (9 rats in each group),respectively,which were control group,hyperthyroidism model group,low,medium and high doses groups (both DIT and KI contents were 25.0,166.7,500.1 μg/kg).Physiological saline was intragastrically administrated to the control group;the hyperthyroidism model group was given thyroid tablet suspension (200.0 mg/kg);DIT and KI groups were given thyroid tablet suspension with corresponding doses of iodine simultaneously.The medicine was given once a day for a mouth,all the rats were sacrificed and heart tissue was collected.The colorimetric method was used to examine the activity of ATPases (Na+-K+-ATPase,Mg2+-ATPase,Ca2+-ATPase).Results The activities of Na+-K+-ATPase,Mg2+-ATPase,Ca2+-ATPase were significantly different statistically between groups (F =2.99,3.03,6.18,all P ＜ 0.01).Compared with the control group [(4.01 ± 0.22),(4.28 ± 0.28),(4.46 ± 0.35) μmol/mg·h],the activities of ATPases (Na+-K+-ATPase,Mg2+-ATPase,Ca2+-ATPase included) were reduced significantly in hyperthyroidism model group [(3.60 ± 0.25),(3.42 ± 0.31),(3.85 ± 0.17)μ mol/mg·h,all P ＜ 0.01];the activities of Mg2+-ATPase in DIT medium dose group [(3.89 ± 0.35)μmol/mg ·h],Ca2+-ATPase in DIT medium and high doses groups [(4.12 ± 0.20),(4.09 ± 0.21)μ mol/mg·h] were reduced significantly (all P ＜ 0.05);the activities of Na+-K+-ATPases,Ca2+-ATPase were decreased significantly in three KI groups [(3.64 ± 0.32),(3.60 ± 0.32),(3.53 ± 0.33),(3.93 ± 0.22),(3.90 ± 0.23),(3.85 ± 0.26)μmol/mg·h],Mg2+-ATPase in KI high dose group [(3.65 ± 0.49)μmol/mg·h] was decreased significantly (P ＜ 0.05or ＜ 0.01).Compared with the hyperthyroidism model group,the activities of ATPase were increased in most of the DIT groups [Mg2+-ATPase in low,medium doses groups:(4.06 ± 0.51),(3.89 ± 0.35)μmol/mg·h;Ca2+-ATPase in low,medium,high doses groups (4.15 ± 0.26),(4.12 ± 0.20),(4.09 ± 0.21)μmol/mg·h,all P ＜ 0.05].Conclusion Supplementation of thyroid tablets in the process of hyperthyroidism formation in Wistar rats will reduce myocardial damage by DTT compared with the same dose of KI.

AIM: To develop a method to determine the content of emodin and chrysophanol in Fuyanling Effervescent Tablets(Radix et Rhizoma Rhei, Flos Lonicerae, Radix Sophorae Tonkinensis, etc.). METHODS: HPLC was used to determine the content of emodin and chrysophanol in Fuyanling Effervescent Tablets. The separation was performed on YWG ODS column with methanol (0.1%) phosphoric acid(85∶15) mixture as a mobile phase and the wavelength of UV detector was at 254nm. RESULTS: The resolution and the linearity of this method was good. in the range of emodin was 50.2～401.6ng( r =0.9999); chrysophanol was 49.9～ 399.2 ng( r =0.9994); and with the average recovery of emodin: 99.4%( RSD =2.2%); chrysophanol:100%( RSD =1.1%). CONCLUSION: The method is simple, rapid and satisfactory and suitable for quality control of Fuyanling Effervescent Tablets.

Objective To Investigation the quality of marrow smear purchased by CDUTCM.Methods The quality and the typ-icality of marrow smears purchased during 2015 -2016 were collectively examined,and then decided whether these smears fit the blood cell morphology experimental teaching requirement.Results Of all the 960 marrow smears purchased these two years, 49.7% failed in smear made or stained,and 16.0% failed to meet the teaching requirements in the typicality of marrow cells.Con-clusion Teaching marrow smears,being different from clinic ones in their preparation and morphological diagnosis,must be of great quality in sustaining and of better typicality in their cell features.

Objective:To study the efficacy and safety of early abdominal puncture drainage (APD) in severe acute pancreatitis (SAP).Methods:A retrospective study was conducted on 189 patients with SAP who were managed at the Department of Intensive Medicine of the Second Affiliated Hospital of Anhui Medical University from January 2013 to May 2020. According to whether ultrasound-guided APD was performed within one week after admission to ICU, these patients were divided into 2 groups: patients treated with APD (the APD group) and patients treated without APD (the non-APD group). Clinical data, including the acute physiological and chronic health status (APACHE) Ⅱ score, modified Marshall score, sequential organ failure evaluation (SOFA) score, and prognostic indicators including the retroperitoneal percutaneous drainage (PCD) rate and length of hospital stay, were compared between the two groups before and 1 week after surgery.Results:Of the 189 SAP patients in this study, there were 110 males and 79 females, aged (52.5±17.4) years old. On admission to ICU, the blood amylase, C-reactive protein, procalcalonin, interleukin-6, APACHE II score, modified Marshall score and SOFA score in the APD group were significantly higher than those in the non-APD group. After 1 week of treatment, most clinical indicators in the 2 groups were significantly improved, and there were no significant differences between these indicators (all P>0.05). There were no significant differences in the abdominal infection, retroperitoneal PCD and mortality rates between the APD group and the non-APD group ( P>0.05). The length of hospital stay [29 (18, 45) vs 21 (15, 32) d] and ICU stay [5 (3, 11) vs. 7 (5, 17) d] in the APD group were significantly higher than those in the non-APD group ( P<0.05). Conclusion:For patients with SAP with peritoneal effusion, early APD effectively improved the condition and prognosis without increasing the peritoneal infection and mortality rates.

Background and purpose:BRCA1/2 plays an important role in maintaining the genome stability.Whether BRCA1/2 germline mutation could increase the tumor sensitivity to radiotherapy,thereby inducing secondary primary cancer after radiotherapy is unclear.This study aimed to investigate whether postoperative radiotherapy is a risk factor for the development of second primary cancer in triple-negative breast cancer(TNBC)patients with BRCA1/2 germline mutation.Methods:This research was based on a previously reported retrospective cohort,i.e.,the Fudan University Shanghai Cancer Center TNBC cohort.Between January 1,2007 and December 31,2014,a total of 292 female TNBC patients with BRCA1/2 mutation were enrolled.We performed logistic regression analysis in patients without BRCA1/2 germline mutation(n=261)and BRCA1/2 germline mutation patients(n=31),respectively,to assess the risk factors affecting the incidence of second primary cancer.We then performed interactive analysis on the above two analyses to evaluate the interactive effect between BRCA1/2 germline mutation and postoperative radiotherapy.P<0.05 indicates a statistically significant difference.The research was approved by Fudan University Shanghai Cancer Center TNBC Ethics Committee(050432-4-2108),and each patient provided written informed consent.Results:Logistic regression analysis in patients with BRCA1/2 germline mutations showed that postoperative radiotherapy significantly increased the risk of secondary primary disease compared to non-radiotherapy[odds ratio(OR)=2.475,95%confidence interval(CI):1.933-3.167,P<0.001].In patients without BRCA1/2 germline mutation,the effect of radiotherapy on the incidence of second primary tumor was not significant.There was a significant interaction between BRCA1/2 germline mutation and postoperative radiotherapy for the incidence of secondary primary cancer(OR=9.710,95%CI:0.320-295.250,P=0.193).Conclusion:Although statistical analysis results show that patients with BRCA1/2 germline mutations have an increased risk of developing a second primary tumor after postoperative radiotherapy compared to patients who have not received radiotherapy,there is no significant correlation between BRCA1/2 germline mutations and radiotherapy for the development of a second primary tumor.Therefore,patients with BRCA1/2 germline mutations who receive radiotherapy after surgery may not increase the risk of developing a second primary tumor.

OBJECTIVE To explore the detoxification mechanism of Terminalia chebula Retz(TCR)soup-processed Euphorbia fischeriana Steud.(EFS)based on LC-MS and simulation processing.METHODS The changes of chemical composition of TCR soup-processed EFS,and dichloromethane extracts of crude EFS after simulation processing with TCR soup were analyzed by LC-MS.Mice were orally administered with alcoholic extracts of crude EFS,alcoholic extracts of water-processed EFS,alcoholic extracts of TCR soup,alcoholic extracts of TCR soup-processed EFS,dichloromethane extracts of crude EFS,dichloromethane extracts of TCR soup-processed EFS,and dichloromethane extracts of crude EFS after simulation processing with TCR soup,respectively.Toxicity changes in TCR soup-processed EFS and dichloromethane extracts of crude EFS after simulation processing with TCR soup were evalu-ated by fecal water contents,release levels of TNF-α and IL-1β,and intestinal pathology.RESULTS A total of 115 and 53 com-pounds were identified in EFS and TCR soup,respectively.The contents of 58 and 12 compounds significantly decreased and signifi-cantly increased respectively in EFS after processing with TCR soup.Compared to the blank control group,the fecal water contents and the release levels of TNF-α and IL-1β significantly increased in both the crude and water-processed EFS groups(P<0.01),and no-table intestinal injury was observed.Compared to the crude EFS group,the fecal water contents and the release levels of TNF-α and IL-1β significantly decreased in both the TCR soup group and the TCR soup-processed EFS group(P<0.01),and repaired intestinal injury was observed.After simulation processing for the dichloromethane extracts of crude EFS with TCR soup,the ion intensity change rates for diterpenoids and tannin phenolic acid compounds ranged from-6.75%to 8.09%and 66.06%to 100.00%,respectively.The ion intensity change rates of diterpenoids in the dichloromethane extracts of TCR soup-processed EFS ranged from-9.92%to 54.72%with almost no tannin phenolic acid compounds.Compared to the blank control group,the fecal water contents and the release levels of TNF-α and IL-1β significantly increased in both the dichloromethane extracts of crude and TCR soup-processed EFS groups(P<0.01),and severe intestinal injury was observed.Compared to the dichloromethane extracts of crude EFS group,the fecal water contents and the release levels of TNF-α and IL-1β significantly decreased in the group of the dichloromethane extracts of crude EFS after simulation processing with TCR soup(P<0.01),with no apparent intestinal injury.CONCLUSION TCR soup pro-cessing can alleviate the intestinal toxicity of EFS.The detoxification mechanism may involve the introduction of a large number of tan-nin phenolic acid compounds in TCR soup during the processing of EFS,which plays a pharmacological antagonistic role in the animal body.