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 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 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 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 To reduce the incidence of falls in elderly patients and improve the quality of nursing safety using tracing method. Methods The tracking methodology was used to formulate fall prevention system and implement tracking path. The tracking centered on patients with high risk of fall and included three aspects: mastery degree of nurses on fall prevention system, participation degree of patients and the implementation of first class quality control. The comparison of tracking result analysis of two quarters in 2016 helped to find out the main reason of the occurrence of falls in elderly patients, make targeted improvement measures and evaluate the effect. Results After the continuous improvement of using tracking methodology, the scores of inspection results and the mastery degree of all nurses on fall core competence in the second quarter were significantly improved when compared to the first quarter (t=23.18, 17.25;P< 0.01); the incidence of falls decreased from 13 cases (6.5%) in the first quarter to 4 cases (2.0%) in the second quarter, while patients' satisfaction degree increased from 85.5% in the first quarter to 98.5% in the second quarter with statistically significant differences between two quarters (χ2=6.38, 14.72; P< 0.01). Conclusions The application of tracing method can effectively reduce the incidence of falls in elderly patients. The method is beneficial to the long-term management of nursing quality, simple, feasible, easy to use, and worthy promoting.

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.

The family readiness for discharge of premature infants in the Neonatal Intensive Care Unit (NICU) is an important index to evaluate the safe discharge of premature infants, and a good family discharge readiness is the basic guarantee for the smooth recovery and healthy growth of premature infants. This article summarizes the concept, influencing factors, and intervention strategies of family discharge readiness for premature infants in NICU, in order to provide reference for the formulation and improvement of discharge readiness measures for premature infants in NICU.

Objective:To evaluate the effect of intermittent fasting on postoperative cognitive function in aged rats and the role of Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway in it.Methods:Eighty SPF healthy male Sprague-Dawley rats, aged 20 months, weighing 600-650 g, were divided into 4 groups ( n=20 each) using a random number table method: control group (group C), postoperative cognitive dysfuction (POCD) group (group P), intermittent fasting + POCD group (group IF+ P), and intermittent fasting+ JAK2/STAT3 signaling pathway agonist C-A1+ POCD group (group IF+ A+ P). IF+ P group and IF+ A+ P group underwent a 4-week intermittent fasting procedure: fasting for 24 h followed by free eating for 24 h, without limiting water intake throughout the entire process, in addition C-A1 100 μg/kg was intraperitoneally injected daily during intermittent fasting in IF+ A+ P group. Rats underwent exploratory laparotomy under sevoflurane anesthesia to prepare POCD models in P group, IF+ P group and IF+ A+ P group. Three days after surgery, open field tests were conducted to evaluate the autonomous motor function of rats, and Morris water maze tests were conducted to evaluate the cognitive function of rats on 3-7 days after surgery. After the Morris water maze tests, the rats were sacrificed and the hippocampal CA1 area was removed for determination of the expression of JAK2, phosphorylated JAK2 (p-JAK2), STAT3, and phosphorylated STAT3 (p-STAT3) (by Western blot) and contents of interleukin-6 (IL-6), IL-1β and tumor necrosis factor-alpha (TNF-α) (by enzyme-linked immunosorbent assay) and for microscopic examination of pathological changes of hippocampal CA1 region (using HE staining). Results:There was no statistically significant difference in the speed, distance and duration of stay at the center of the open field test among the four groups ( P>0.05). Compared with group C, the escape latency was significantly prolonged, the number of crossing the original platform was reduced, and the p-JAK2/JAK2 ratio, p-STAT3/STAT3 ratio and contents of IL-6, IL-1β and TNF-α in the hippocampal CA1 region were increased ( P<0.05), and the pathological damage occurred in the hippocampal CA1 region in group P. Compared with P group, the escape latency was significantly shortened, the number of crossing the original platform was increased, and the p-JAK2/JAK2 ratio, p-STAT3/STAT3 ratio and contents of IL-6, IL-1β and TNF-α in the hippocampal CA1 region were decreased ( P<0.05), and the pathological damage in the hippocampal CA1 area was significantly alleviated in IF+ P group. Compared with IF+ P group, the escape latency was significantly prolonged, the number of crossing the original platform was reduced, and the p-JAK2/JAK2 ratio, p-STAT3/STAT3 ratio and contents of IL-6, IL-1β and TNF-α in the hippocampal CA1 region were increased ( P<0.05), and the pathological damage in the hippocampal CA1 area was aggravated in group IF+ A+ P. Conclusions:Intermittent fasting can improve postoperative cognitive function in aged rats, and the mechanism may be related to inhibiting the JAK2/STAT3 signaling pathway and reducing the neuroinflammatory responses in the hippocampal CA1 region.

Objective:To investigate the application of patient-based real-time quality control (PBRTQC) using exponentially weighted moving average (EWMA) method in internal quality control (IQC) procedures of thyroid function tests.Methods:The serum thyroid function test results of outpatients and inpatients in the Second Hospital of Shandong University from December 1, 2022 to April 30, 2023 were collected. Based on the PBRTQC professional intelligent software system, normality correction, parameter setting, program preparation and real-time operation of test data were carried out. The results of all patients who underwent thyroid function testing between May 1, 2023 and August 31, 2023 were used as the validation dataset. The estimated EWMA value of thyroid function test results and the cumulative coefficient of variation ( CV) over 4 months were calculated. The cumulative CV was compared with the criteria of precision quality standard (1/3TEa) and the CV of IQC. Westgard 2-2s and 1-3s rules were used for alarm setting. The early warning information of the EWMA quality control program were recorded and the potential causes of performance changes were analyzed. DxLab Mind software was used to conduct normal distribution statistics for all data, and the Kolmogorov-Smirnov test was performed on the test results. Results:The items related to serum thyroid function of the patients were all positively skewed. After data correction by Box-Cox method, the PBRTQC data of free triiodothyronine (FT3) and free thyroxine (FT4) were normally distributed, and their cumulative precisions ( CV) of EWMA within 4 months were 6.26% and 2.86%, respectively, both of which were lower than the precision quality target of 8.33%. However, the data of thyroid-stimulating hormone (TSH), thyroid peroxidase antibodies (TPOAb) and thyroglobulin antibody (TgAb) were still positive skewed after modification. The EWMA cumulative CV of TSH, TPOAb and TgAb were 13.16%, 15.31% and 16.77%, which were higher than the precision quality targets of 8.33%, 10% and 10%, respectively. The EWMA QC program can detect different out-of-control alarms, including FT4 false alarms due to sample source concentration and TSH result bias caused by changes in reagent performance. In addition, the EWMA QC program can also detect differences in FT3 results between different DXI800 fully automated chemiluminescence instrument instruments. Conclusions:The EWMA program based on PBRTQC professional intelligent software tools can monitor the patient data of the detection system in real time and continuously, dynamically identify and monitor the errors generated during the analysis process and give early warning. It can be used as a useful supplement for the daily IQC of thyroid function items, especially FT3 and FT4, and has good clinical application value.