Objectives:Aimed to establish a rapid, high-throughput, automated method for determining the creatine kinase (CK-MM) isoform levels.Methods:Magnetic beads labeled with anti-CK-MM antibodies were combined with alkaline phosphatase-based chemiluminescence detection. Clinical and diagnostic performance validation of the assay was determined by analysis of 998 and 75 dried blood spot samples from healthy newborns and Duchenne muscular dystrophy (DMD) patients, respectively, and the CK activity was also determined. The blank and detection limits, cross-reactivity, recovery rate of the method, intra-and inter-assay coefficient, and the hook effect were evaluated.Results:Blank and detection limits were 17.4 and 39.3 ng/ml, respectively. Cross-reactivity toward CK-MB and CK-BB isoforms was 0.2% and 0.02%, respectively. Intra-and inter-assay coefficients of variation were<1%. Mean recovery was 100.32%, with no hook effect in CK-MM levels<50 000 ng/ml. Overall, the mean CK-MM concentrations in newborns and DMD patients were (27.05±0.97) and (3 720±300.5) ng/ml, respectively. A significant positive correlation between the dried blood spot detected CK-MM levels and total CK enzyme activity, evaluated in corresponding serum samples from the 75 DMD patients, was observed ( r=0.91, P<0.001), ?which is in good agreement with the clinical. Conclusions:An assay for rapid quantitative determination of CK-MM that meets clinical newborn screening requirements was established. It had a good value for application.

ObjectiveTo study the differences in volatile oil content of bran-processed Atractylodes lancea and its standard decoction concentrate and freeze-dried powder， as well as the differences in the types and contents of chemical components in volatile oil， and to clarify the quality value transmitting. MethodTen batches of A. lancea rhizoma were collected and prepared into raw products and bran-processed products of A. lancea， standard decoction concentrate and freeze-dried powder of bran-processed A. lancea in order to extract the volatile oil， and the transfer rate of volatile oil in each sample was calculated. Quantitative analysis of the main chemical components（β-eudesmol， atractylon， atractylodin） in each volatile oil was performed by gas chromatography（GC） on the HP-5 quartz capillary column（0.32 mm×30 m， 0.25 μm） with a flame ionization detector（FID）， a split ratio of 10∶1 and a temperature program（initial temperature at 80 ℃， hold for 1 min， rise to 150 ℃ at 10 ℃·min^-1， hold for 10 min， rise to 155 ℃ at 0.5 ℃·min^-1， hold for 5 min， rise to 240 ℃ at 8.5 ℃·min^-1， hold for 8 min）. Cluster analysis and principal component analysis（PCA） were used to explore the overall differences in types and contents of chemical components between the standard decoction concentrate and freeze-dried powder. ResultThe transfer rates of volatile oil in the bran-processed products， standard decoction concentrate and freeze-dried powder were 70.51%， 1.57% and 40.90%， respectively. The average transfer rates of β-eudesmol， atractylon and atractylodin in the volatile oil of bran-processed A. lancea were 58.45%， 48.49% and 55.64%， respectively. In the standard decoction concentrate， only β-eudesmol and atractylodin were detected， and their average transfer rates were 0.22% and 0.10%， respectively. And only β-eudesmol was detected in the freeze-dried powder with the average transfer rate of 8.37%. The results of cluster analysis and PCA showed that there are obvious differences in the types and contents of chemical components between the standard decoction concentrate and freeze-dried powder. ConclusionThe quality value transmitting between bran-processed A. lancea and its standard decoction concentrate and freeze-dried powder is stable， and if the freeze-dried powder is selected as the reference material of dispensing granules， appropriate amount of volatile oil should be added back to make it consistent with the quality of the standard decoction concentrate.

Objective:To develop a novel, flexible, dual-arm, master-slave digestive endoscopic minimally invasive surgical robot system named dual-arm robotic endoscopic assistant for minimally invasive surgery (DREAMS) and to evaluate its feasibility for endoscopic submucosal dissection (ESD) by using ex vivo porcine stomachs.Methods:A novel endoscopic robot (DREAMS) system was developed which was composed of a flexible two-channel endoscope, two flexible robotic manipulators, a master controller, a robotic arm, and a control system. A total of 10 artificial round-like lesions with diameters ranging from 15 to 25 mm were created (5 in gastric antrum and 5 in gastric body) by using fresh peeled stomach of healthy pigs as the model. Submucosal dissection was performed with the assistance of the DREAMS system by two operators. The main outcome was submucosal dissection speed, and the secondary outcomes included muscular injury rate, perforation rate, and grasping efficiency of the robot.Results:All 10 lesions were successfully dissected en bloc by using the DREAMS system. The diameter of the artificial lesions was 22.34±2.39 mm, dissection time was 15.00±8.90 min, submucosal dissection speed was 141.79±79.12 mm 2/min, and the number of tractions required by each ESD was 4.2 times. Muscular injury occurred in 4/10 cases of ESD. No perforation occurred. Conclusion:The initial animal experiment shows the DREAMS system is safe and effective.

Objective:To investigate the influence of DTAS gene mutations (DNMT3A, TET2, ASXL1, SRSF2) on survival of newly diagnosed acute myeloid leukemia (AML) patients.Methods:A retrospective analysis of 163 patients with next-generation sequencing(NGS)data in the hematology Department of Affiliated Hospital of Xuzhou Medical University from January 1, 2018 to October 31, 2021 was performed. Clinical data of patients were collected and analyzed including patient′s height, weight, gender, age, bone marrowblast ratio, induction chemotherapy regimen, transplantation or not, complete blood count, etc. There were 88 males and 75 females with a median age of 48 (4-81) years. According to the next-generation sequencing data, patients with any mutation of the above four genes were divided into the DTAS gene mutation group, and vice versa, they were divides into non-DTAS gene mutation group.The Kaplan-Meier method and Cox proportional risk model were used to analyze survival and prognosis.Results:Among 163 patients, DTAS gene mutation was detected in 50 patients (30.7%), and not in 113patients(69.3%). Among the 50 patients with DTAS genemutations, 8 cases(16%) had≥2 mutations and 42 cases(84%) had any gene mutation in DTAS. In the DTAS gene mutation group, the patients were older, the stratification of the European leukemia network(ELN) was poor, the duration of remission was shorter, the proportion of sever myelosuppression degree was higher (61.8% vs 34.8%) at day28 after induction chemotherapy, and the lymphocyte-monocyte ratio was lower than that of the healthy control group when CR was reached after treatment.The results of K-M survival analysis showed that the overall survival(OS)time ( P=0.003) and event-free survival(EFS) ( P=0.008) time of the DTAS gene mutant were significantly shorter than those of the non-DTAS gene mutated group.The medianOS timein theh DTAS gene mutations was significantly shorter than that in the non-DTAS gene mutated group ( P=0.003, HR=2.041) [21(95% CI 16.63-25.37) months vs 43 (95% CI 33.01-52.99) months].The results of multivariate COX analysis revealed that DTAS gene mutations was an independent risk facror forOS time(HR=2.041, 95% CI: 1.285-3.244, P=0.003) in AML patients. Conclusion:DTAS gene mutation does not affect the hematopoietic recovery time after induction chemotherapy, but the duration of remission is shorter in the DTAS gene mutations group. DTAS gene mutations indicate a poor prognosis, which is an independent risk factor for OS.

Objective:To construct the core indicators for entrustable professional activities in specialists in obstetrics and gynecology.Methods:A study group was formed by the specialists in obstetrics and gynecology and the experts in medical education. The core indicators for entrustable professional activities were constructed for the specialists in obstetrics and gynecology based on literature review and clinical practice, and then the Delphi method was used to conduct two rounds of expert letter consultation for screening and optimization from March 2021 to January 2023 to further identify the core indications.Results:The expert positive coefficient was 100% for the two rounds of consultation, with an expert authority coefficient of 0.82, and the Kendall's coefficient of concordance was 0.221 and 0.213, respectively (both P<0.01). Ten core indicators and their content descriptions were constructed for entrustable professional activities in obstetrics and gynecology specialists, and the experts had a degree of recognition of more than 80% for the importance of these ten entrustable professional activities, with a coefficient of variation of <0.25. This study determined the expected entrustable level of each indicator for specialists at the completion of the course, which ranged from grade 3 to 5; the highest level of 4.48 was observed for the diagnosis and treatment of outpatients, which was between the levels of mastery and expert; the lowest level of 3.52 was observed for laparoscopic hysterectomy, which was between the levels of competency and mastery. Conclusion:This study preliminarily constructs the core indicators for entrustable professional activities in specialists in obstetrics and gynecology, which provides a new exploration for the standardized training of specialists.

Objective:To analyze the change trend of survival rate after hip fragility fracture in Tianjin Hospital from 2015 to 2021, and the influence of gender, marital status, age and number of complications on survival rate.Methods:From January 1, 2015 to December 31, 2021, a total of 12,570 patients with fragility fracture of hip were retrieved, including 3,934 males and 8,636 females. The age at admission was 74.11±9.50 years and 74.62±8.99 years respectively. By comparing the ID number with the Tianjin population information database, 2,054 cases died, including 804 males and 1,250 females, aged 81.34±7.88 years and 81.92±7.42 years respectively at the time of death. Acquire the patient's survival status, calculate the cumulative survival rate at 3 month intervals, study the change rule of the cumulative survival rate over time, and use Kaplan-Meier method to calculate the cumulative survival rate of the whole population and the impact of gender, marital status, age, and number of complications on the survival rate.Results:The median survival time of all the dead people after fracture was 13(3, 31) months, including 11(2, 27) months for males and 15(4, 32) months for females. Joinpoint regression showed that 9 months after the hip fragility fracture was the break point of the survival rate. The mortality rate changed significantly within 9 months after fracture (the annual change rate was 47%), and slowed down 9 months later (the annual change rate was 1%). There was a statistically significant difference in trend detection before and after the break point ( P<0.05). The age at admission was 80.11±7.71 years for the dead and 73.36±9.01 years for the non dead, with a statistically significant difference ( t=31.80, P<0.001). After normalization, the number of complications was 0.20±0.93 among the dead and 0.00±0.87 among the non dead, with a statistically significant difference ( t=88.81, P<0.001). The survival rate of men after hip fracture is lower than that of women, the number of people without spouse is lower than that of people with spouse, the number of people with more than 70 years old is lower than that of people with less than 70 years old, and the number of complications ≥2 people is lower than that of people with less than 1 complication, all of which are statistically significant ( P<0.05). Conclusion:The survival rate within 9 months after the occurrence of hip fragility fracture decreased significantly, and it needs to be tracked and managed for at least 9 months to effectively reduce the risk of death; Male, no spouse, age>70 years old, number of complications ≥2 will increase the risk of death after hip fragility fracture, leading to reduced survival rate of patients.

Objective:To automatically synthesize Al 18F-1, 4, 7-triazacyclononane-1, 4, 7-triacetic acid (NOTA)-fibroblast activation protein inhibitor (FAPI)-04, perform PET/CT imaging in vivo, and evaluate its diagnostic efficacy on tumors. Methods:Al 18F-NOTA-FAPI-04 was produced in All-in-one automatic synthesis module and its quality control was conducted by high performance liquid chromatography (HPLC) equipped with a radioactive detector. Al 18F-NOTA-FAPI-04 PET/CT imaging was performed in normal BALB/c mice ( n=3) and 4T1 breast cancer models ( n=3) to determine its biodistribution. Then Al 18F-NOTA-FAPI-04 and 18F-FDG PET/CT imaging were performed in a hepatocellular carcinoma patient (male, 51 years old). Results:The synthesis time of Al 18F-NOTA-FAPI-04 was about 35 min, and the radiochemical yield was (25.2±1.9)% (attenuation correction, n=3). The product was colorless transparent solution with pH value of 7.0-7.5, and the specific activity was (46.11±3.07) GBq/μmol (attenuation correction, n=3). The radiochemical purity was above 99.0% and was still above 98.0% at room temperature after 6 h. PET/CT imaging in mice showed that physiological uptake of Al 18F-NOTA-FAPI-04 was mainly in biliary system and bladder, and Al 18F-NOTA-FAPI-04 highly concentrated in tumor xenografts. PET/CT imaging in the patient showed that Al 18F-NOTA-FAPI-04 obtained high tumor background ratio (TBR) values of 4.1, 8.9, 5.4, 4.8, 2.2 in parasternal lymph nodes, anterior diaphragmatic lymph nodes, hilar lymph nodes, pancreaticoduodenal ligament lymph nodes, abdominal aortic lymph nodes, respectively, while TBR values were 1.0, 2.8, 5.0, 2.1, 1.1 by 18F-FDG. Conclusions:Al 18F-NOTA-FAPI-04 can be synthesized with short time, high radiochemical yield and good stability using All-in-one module. Al 18F-NOTA-FAPI-04 PET/CT imaging has high contrast and excellent diagnostic efficacy on tumors.

Objective : To explore the effect of liver-specific knockout of transcription factor EB (Tfeb) on high-fat diet ( HFD ) -induced hepatic steatosis in mice． Methods : Wild-type C57BL /6J mice and liver-specific Tfeb knockout C57BL /6J mice were fed with HFD or normal chow for 12 weeks，respectively，and then the serum tri- glyceride (TG) ，total cholesterol ( TC) ，monocyte chemoattractant protein-1 ( MCP-1 ) ，tumor necrosis factor-α (TNF-α) ，aspartate aminotransferase ( AST) and alanine transaminase ( ALT) were measured in each group of mice ; Western blot was used to detect Tfeb protein expression levels in liver tissues of mice in each group，HE stai- ning was used to monitor histopathological changes in liver tissues of mice in each group，oil red O staining was used to monitor lipid deposition in liver tissues of mice in each group，and F4 /80 fluorescence staining was used to monitor macrophage infiltration in liver tissues of mice in each group. Results : There was no expression of Tfeb gene in liver of liver-specific knockout Tfeb mice，suggesting that the effect of Tfeb gene knockout in liver tissue was better．Compared with the normal control group，the expression of Tfeb protein in the liver tissue of the model control group was down-regulated．Compared with the normal control group，both the HE staining results and the oil red O staining results showed that the liver specific Tfeb caused lipid deposition and liver lobule disorder in mice， which was similar to the liver changes in the model control mice，however，liver-specific knockout of Tfeb mice at 12 weeks of HFD had more severe liver lipid deposition and hepatic lobular structural disorder．The results of F4 / 80 fluorescence staining indicated that the specific knockout of liver Tfeb could aggravate the infiltration of macro- phages in the liver of mice induced by high-fat feeding．At the same time，the serological test results indicated that compared with the normal control group，the serum levels of TC，TG，TNF-α , MCP-1，AST and ALT in the liver- specific knockout Tfeb group and the model control group increased ，and these changes were further elevated in Tfeb knockout mice after HFD feeding． Conclusion Liver-specific knockout of Tfeb aggravates HFD-induced he- patic steatosis in mice.

Objective:To observe the effect of combining scalp acupuncture with hyperbaric oxygen on serum homocysteine and highly-sensitive c-reactive protein levels and functional recovery after cerebral infarction.Methods:A total of 101 survivors of cerebral infarction were divided randomly into a scalp acupuncture group ( n=33), a hyperbaric oxygen group ( n=34) and a combined treatment group ( n=34). All received routine treatment plus the appropriate supplementary treatment once a day for 10 days. The subjects were evaluated before the experiment as well as 10 and 90 days afterward. The National Institutes of Health′s stroke scale (NIHSS) was used to quantify neurological deficits and the Barthel Index quantified ability in the activities of daily living. Ninety days after the treatment, modified Rankin scale scores were also assigned. The levels of serum homocysteine (Hcy) and highly-sensitive c-reactive protein (hs-CRP) before and after 10 days of treatment were also compared among the 3 groups. Results:The average NIHSS and Barthel Index scores of all three groups had improved significantly after 10 days of treatment and the improvements persisted at the follow-up 3 months later. Both results were significantly better in the combined treatment group than in the scalp acupuncture group at the 90-day follow-up evaluation. The average Rankin score of the combined treatment group was lower at the last follow-up. Compared with before the intervention, the average Hcy of the scalp acupuncture group, the average hs-CRP of the hyperbaric oxygen group, as well as the average Hcy and hs-CRP of the combined treatment group were significantly lower after 10 days of treatment. Compared with the scalp acupuncture group, the average Hcy [(11.68±2.6) μmol/L] of the combined treatment group was significantly lower after the 10 days of treatment.Conclusions:Supplementing scalp acupuncture with hyperbaric oxygen therapy improves the long-term outcomes of cerebral infarction, reducing the level of serum Hcy.

AIM: To evaluate the effect of TDF withdrawal time on changes of serum HBV-M, HBV DNA and ALT level in the mother-to-child blocking of the maternal population. METHODS: A prospective, randomized and controlled study was conducted. The 120 pregnant women with HBV who took TDF during 24 to 28 weeks of gestation were randomly divided into group A (withdrawal at delivery) and group B (withdrawal at 4 weeks postpartum), levels of HBV-M, HBV DNA, and ALT at different times were detected. The results were statistically analyzed by Wilcoxon Rank-sum test and χ