To accurately analyze metabolites in industry-important photosynthetic microbes, LC-MS based metabolomics protocol needs to be optimized specifically for individual species. In this study, an LC-MS based metabolomics method was optimized for cyanobacterium Synechocystis sp. PCC 6803. With the optimized extraction, liquid chromatographic and mass spectral parameters, the method was capable of detecting 24 important metabolites related to central carbohydrate and energy metabolism in Synechocystis sp. PCC 6803. The study laid an important foundation for the metabolomics analysis of cyanobacteria.
Chromatography, Liquid ; Mass Spectrometry ; Metabolome ; Metabolomics ; Photosynthesis ; Synechocystis ; metabolism

BACKGROUND:Toxic cytarabine is often used to prepare highly purified neurons in experimental studies addressing central nervous system diseases. However, the intervention time of cytarabine is little reported. OBJECTIVE:To determine the optimal intervention time of cytarabine(final concentration 10μmol/L) in primary culture of rat cortical neurons. METHODS:Rat primary cortical neurons were cultured in Neurobasal+B27 medium, and 10μmol/L cytarabine were added at 12, 24, 36 and 48 hours after culture, respectively. Half of the medium were changed every 48 hours. The morphology of neurons was observed under inverted microscope at 7 days. The purity and differentiation of neurons maturity were identified by immunocytochemistry method of neuron specific enolization enzyme staining. Morphometric analysis for all neuron-specific enolase positive cells was performed by Multifunction Computer Image Analysis System. RESULTS AND CONCLUSION:After addition of cytarabine at 24 hours of culture, the purity of neurons was more than 90%, well-differentiated cortical neurons accounted for 89.00%, and the area of neuronal body was the largest with the longest synapses. There were more neuron cells with transparent cytoplasm, and large nucleus. The cell body had good refraction and strong stereo sense. The neurons with 3-4 synapses and 3-4 bifurcates formed a good network structure. These results illustrate that although it willbe beneficial for the purity of neurons to add cytarabine early in neuron culture process, it will make obvious effect on neuronal differentiation. The highly purified and well-grown cerebral cortical neurons will be obtained after cultured in neurolbasal medium, which cytarabine is added to at 24 hours of culture.

Objective To determine the toxicity of therapeutical Serratia marcescans vaccine when repeated intracerebral administration into rat brain.Methods SD rats are prepared by intracranial embedding location catheter and were randomly divided into 8 groups:namely normal control,lunar control group(give NS in same dose),low dosage group,middle dosage group and high dosage group of acute stage or restore stage.Three dosage of vaccine S311 were administrated(low 320 million/kg,middle 1600 million/kg,high 8000 million/kg).The embedding catheter rats were fixed point injecting vaccine,once per day for 15 days with microsyringe of microdialysis device.While continuously record the common status,appetite,body weight of animals.25 days later,Animals were killed to observe the morphology of brain.Results The main pathologic changes of high dosage group were inflammatory cell infiltration into the tissues around injecting location,subarachnoid space,and ependyma.The inflammatory cell is mainly gial cell,monocytes,lymphocytes.No degeneration and necrosis of brain tissue were observed.The inflammatory reaction of brain tissues around injecting location was correlated with the dosages.Except the inflammation around injecting location,the other brain tissues were normal and absent of organic pathological changes.After 25 days restoration,the inflammation around injecting location was absorbed.Conclusions The method of intracranial embedding catheter and fixed point injecting is successful.Intracranial administration of therapeutical Serratia marcescans vaccine is mainly effect on location around injecting to elicit localized,reversible,and non-specific inflammatory reaction.

Objective: To determine the value of telomerase activity in urine exfoliated cells as clinical indicator of tumor presence, stage, and recurrence. Methods: The techniques of TRAP-PCR and TRAP-sliver staining were employed to detect telomerase activity in 73 patients with TCC before operation (study group) and 20 benign urothelial cancer patients (control group) and 21 normal individuals (normal group). Cytologic results were obtained simultaneously. The bladder tumor specimens were obtained from 73 patients during operation, and histologically evaluated for tumor content and grade. Results: Positive rate of telomerase activity detection in TCC patients (80.8%,59 of 73) was significantly higher than that of cytological examination (20.5%, 15 of 73). Positive telomerase activity was not found in 17 of 20 in control group and none was found in normal group. Analysis of the distribution of abnormalities with tumor stage revealed greater detection of high pathological stage (T 2-T 4) (89.7%, 26 of 29) compared with low stage (Tis-T 1) (75%, 33 of 44). Conclusion: Detection of telomerase activity in urine exfoliated cells may be a sensitive and effective marker in the diagnosis of TCC.

Objective To determine the value of detection of H ras oncogene mutation in urine exfoliated cells as clinical indicator of tumor presence, recurrence and stage.Methods Point mutation at codon 12 of H ras gene was assayed by polymerase chain reaction followed by analysis of single strand conformation polymorphism in urine exfoliated cells from 48 patients with transitional cell carcinoma before operation and 28 patients with non urothelial cancer or normal individuals. The mutation was further confirmed by dideoxy mediated chain termination method of DNA sequencing. Cytology analysis was carried out simultaneously. Bladder tumor specimens were obtained from 48 patients during operation, and histologically elevated for tumor content and grading.Results 48%(23 of 48) of the patients were detected by their aberrant band in SSCP. All aberrant bands displayed a mutant H ras sequence, where 15% (7 of 48) of the patients displayed, apositive cytological analysis. Analysis of abnormalities with tumor stage revealed that the greater detection of high pathological stage (Ⅲ Ⅳ) compared with low stage (Ⅰ Ⅱ) was related to the recurrence of transitional cell carcinoma.Conclusion Our results suggest that the detection of H ras mutations may be of clinical value in the detection of TCC.

Objective:Comparison of early cholecystectomy (EC) and delayed cholecystectomy (DC) after percutaneous transhepatic gallbladder drainage (PTGD) for over 80-year-old patients with acute cholecystitis.Methods:Clinical data of 297 over-80-year-old patients of with acute cholecystitis undergoing surgery in Shidong Hospital Affiliated to the University of Shanghai for Science and Technology from January 2016 to January 2023 were retrospectively analyzed, including 123 males and 174 females, aged (86.1±5.2) years. There were 176 cases in EC group and 121 in PTGD-DC group. Demographic data and perioperative outcomes were compared between the groups, including gender, age, ASA score, lab test, grades of acute cholecystitis, symptom, time before EC or PTGD, intraoperative blood loss, conversion, respiratory disfunction, gangrenous cholecystitis, abdominal drainage time, postoperative complication, hospital stay, intensive care time.Results:The baseline characteristics were similar between EC and PTGD-DC groups, including demographics, ASA score, grades of acute cholecystitis, white blood cell counting, platelet counting, level of serum procalcitonin, time before EC or PTGD, and percentage of comorbidity. Compared to PTGD-DC group, Patients in DC group experienced more intraoperative blood loss (118±62 vs 32±31ml], longer operative time (135±43 vs 61±31) min], higher incidence of gangrenouscholecystitis [23.2%(41/176) vs 9.9%(12/121)], more respiratory support [16.5%(29/176) vs 11.6%(14/121)], more conversion to open surgery [22.7%(40/176) vs 9.1%(11/121)], longer postoperative abdominal drainage time (9.1±2.6 vs 3.8±2.3d], longer hospitalization (8.2±3.1 vs 6.1±2.2 d], longer intensive care (9.0±0.3 vs 4.6±0.2 h] (all P<0.05). More complications were observed in EC group, such as bile leakage, postoperative bleeding, unscheduled reoperation, bile duct injury (all P<0.05). Conclusion:PTGD and DC could lower the perioperative risk of elderly patients with acute cholecystitis.

With in-depth research and development of dermoscopy, the dermoscopic features including perifollicular pigments, perilesional pigments, pigment network structure, satellite phenomenon and "tapioca sago" appearance, micro-Koebner phenomenon and comet tail-like phenomenon have provided a basis for the evaluation of vitiligo activity. This review summarizes progress in the evaluation of vitiligo activity with dermoscopy in recent years, aiming to promote the application of dermoscopy in the assessment of vitiligo activity.

Objective To explore the expression level of visceral adipose tissue-derived serine protease inhibitor(Vaspin)and secreted frizzled-related protein5(SFRP5)in the serum of children with idiopathic short stature(ISS)and its diagnostic value.Methods 70 children with ISS diagnosed in the First Hospital of Zhangjiakou from December 2021 to February 2023 were selected as the disease group,while 72 healthy volunteer children who underwent physical examination were collected as the control group.Immunoluminescence was applied to detect the expression level of VASPIN,Enzyme-linked immunosorbent assay(ELISA)was applied to detect the expression level of SFRP5 the clinical data of children in two groups were analyzed.Receiver operating characteristic(ROC)curve was applied to analyze the diagnostic value of serum Vaspin and SFRP5 for ISS,multivariate Logistic regression was used to analyze the influencing factors of ISS.Results Compared with the control group,the serum Vaspin level in the disease group was obviously increased(2.89±0.92 ng/ml vs 1.81±0.42 ng/ml),while the SFRP5 level was obviously reduced(10.22±2.84 pg/ml vs 13.21±3.53 pg/ml),the differences were statistically significant(t=9.040,5.552,all P＜0.05).The weight,height,body mass index(BMI)and proportion of sexual development stage II～V of children in the disease group were obviously lower than those in the control group,and the differences were statistically significant(t=7.687,6.330,5.559,7.024,all P＜0.05).The area under ROC curve showed that the AUC of Vaspin and SFRP5 and their combined detection in the diagnosis of ISS were 0.768,0.849 and 0.925,respectively,the combined diagnosis efficacy of Vaspin and SFRP5 was better than that of serum Vaspin and SFRP5 alone(Z =3.829,P＜0.001;Z =2.141,P=0.032).Multivariate Logistic regression analysis showed that BMI(OR=0.508,95%CI:0.260～0.991),Vaspin(OR=3.458,95%CI:1.125～10.631)and SFRP5(OR=0.378,95%CI:0.153～0.935)were the influencing factors for ISS(all P＜0.05).Conclusion The expression level of Vaspin in the serum of children with ISS is obviously increased,while the expression level of SFRP5 is obviously reduced.The two are influencing factors of ISS,and the combined detection of their expression levels has certain value in the diagnosis of ISS.

Objective: To investigate the current status and real performance of the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels in China through interlaboratory comparison. Methods: Peking University People’s Hospital (PKUPH) prepared the samples for comparison. That is, the fresh RUNX1-RUNX1T1 positive (+) bone morrow nucleated cells were serially diluted with RUNX1-RUNX1T1 negative (-) nucleated cells from different patients. Totally 23 sets with 14 different samples per set were prepared. TRIzol reagent was added in each tube and thoroughly mixed with cells for homogenization. Each laboratory simultaneously tested RUNX1-RUNX1T1 and WT1 transcript levels of one set of samples by real-time quantitative PCR method. All transcript levels were reported as the percentage of RUNX1-RUNX1T1 or WT1 transcript copies/ABL copies. Spearman correlation coefficient between the reported transcript levels of each participated laboratory and those of PKUPH was calculated. Results: ①RUNX1-RUNX1T1 comparison: 9 samples were (+) and 5 were (-) , the false negative and positive rates of the 20 participated laboratories were 0 (0/180) and 5% (5/100) , respectively. The reported transcript levels of all 9 positive samples were different among laboratories. The median reported transcript levels of 9 positive samples were from 0.060% to 176.7%, which covered 3.5-log. The ratios of each sample’s highest to the lowest reported transcript levels were from 5.5 to 12.3 (one result which obviously deviated from other laboratories’ results was not included) , 85% (17/20) of the laboratories had correlation coefficient ≥0.98. ②WT1 comparison: The median reported transcript levels of all 14 samples were from 0.17% to 67.6%, which covered 2.6-log. The ratios of each sample’s highest to the lowest reported transcript levels were from 5.3-13.7, 62% (13/21) of the laboratories had correlation coefficient ≥0.98. ③ The relative relationship of the reported RUNX1-RUNX1T1 transcript levels between the participants and PKUPH was not always consistent with that of WT1 transcript levels. Both RUNX1-RUNX1T1 and WT1 transcript levels from 2 and 7 laboratories were individually lower than and higher than those of PKUPH, whereas for the rest 11 laboratories, one transcript level was higher than and the other was lower than that of PKUPH. Conclusion The reported RUNX1-RUNX1T1 and WT1 transcript levels were different among laboratories for the same sample. Most of the participated laboratories reported highly consistent result with that of PKUPH. The relationship between laboratories of the different transcript levels may not be the same.