A derivatizaton method combined with gas chromatography-mass spectrometry(GC-MS)was established for detection of isobutyl chloroformate(IBCF)residue in active pharmaceutical ingredient of agatroban.The extraction and derivatization reagents,derivatization time,qualitative and quantitative ions were selected and optimized,respectively.The possible mechanism of derivatization and characteristic fragment ions fragmentation were speculated.The agatroban samples were dissolved and extracted by methanol,and the residual IBCF was derived with methanol to generate methyl isobutyl carbonate(MIBCB).After 24 h static derivatization at room temperature,IBCF was completely transformed into MIBCB,which could be used to indirectly detect IBCF accurately.The results showed that the linearity of this method was good in the range of 25-500 ng/mL(R2=0.9999).The limit of detection(LOD,S/N=3)was 0.75 μg/g,and the limit of quantification(LOQ,S/N=10)was 2.50 μg/g.Good recoveries(95.2%-97.8%)and relative standard deviations(RSDs)less than 3.1%(n=6)were obtained from agatroban samples at three spiked levels of IBCF(2.50,25.00,50.00 μg/g),which showed good accuracy of this method.Good precision of detection results was obtained by different laboratory technicians at different times,the mean value of spiked sample solution(25.00 μg/g)was 24.28 μg/g,and the RSD was 2.1%(n=12).The durability was good,minor changes of detection conditions had little effect on the results.Under the original condition and conditions with initial column temperature±5℃,heating rate±2℃/min,column flow rate±0.1 mL/min,the IBCF content of spiked sample solution(25.00 μg/g)was detected,the mean value of detection results was 24.16 μg/g,and the RSD was 2.2%(n=7).Eight batches of agatroban samples from two manufacturers were detected using the established method,and the results showed that no IBCF residue was detected in any of these samples.The agatroban samples could be dissolved by methanol,and then the IBCF residue could be simultaneously extracted and derived with methanol as well.This detection method had the advantages of simple operation,high sensitivity,low matrix effect and accurate quantification,which provided a new effective method for detection of IBCF residue in agatroban.

Objective:To study the clinical characteristics and risk factors of nephrocalcinosis in preterm infants.Methods:From March 2021 to August 2021, all preterm infants admitted to NICU of our hospital were retrospectively analyzed. The infants were assigned into nephrocalcinosis group and non-nephrocalcinosis group according to urinary tract ultrasound. Clinical data including gestational age, birth weight(BW), nutritional support strategy and complications were reviewed.Results:A total of 40 preterm infants (<34 weeks) were enrolled. 9 cases were in the nephrocalcinosis group and 31 cases in the non-nephrocalcinosis group. The nephrocalcinosis group had lower BW[(1 167±214) g vs.(1 586±215) g], higher calcium [6.9 (5.1, 8.7) g vs.3.3 (2.1, 6.8) g] and vitamin D intake [3.2(2.5, 4.2)×10 4U vs.1.7(1.1, 3.2)×10 4U] during hospitalization. No significant differences existed between the two groups on the following items:blood calcium and phosphate, 25-hydroxyvitamin D, feeding strategy, time to reach full enteral feeding(TFF), furosemide dosage and respiratory support duration ( P>0.05). In the nephrocalcinosis group, the median age of diagnosing nephrocalcinosis was 40.0(30.0, 52.5)d after birth. 5 cases showed bilateral nephrocalcinosis. 5 cases in the nephrocalcinosis group received renal tubule function examination,4 cases had increased urine β2 microglobulin and 2 cases had increased urine α1 microglobulin. 7 cases had elevated urine calcium in the nephrocalcinosis group. Follow-up showed that nephrocalcinosis disappeared 3-9 months after birth. Conclusions:BW, total calcium and vitamin D intake are risk factors for nephrocalcinosis in preterm infants. Increased urine β2 microglobulin and calcium levels are common co-morbidities in preterm infants with nephrocalcinosis.

Using bioinformatics methods to predict the physical and chemical properties,spatial structure,and T and B cell epitopes of Brucella Periplasmic Protein(EipB),and to explore the immune effects of T and B dominant epitope peptides,for Brucella Periplasmic Protein(EipB)Lay the foundation for the screening of molecular peptide vaccine against bacillosis.Online websites and bioinformatics-related databases were used to analyse the physicochemical properties,transmembrane structural domains,signal peptide sequences,secondary and tertiary structures,hydrophilicity/hydrophobicity and T and B antigenic epitopes of the EipB protein.And select the B and T epitopes with the highest antigenicity(located at 121-137 and 95-110 respectively)to connect with hemocyanin,and after immunizing the mice,detect the production of IgG,IgM,IgA and IgE an-tibodies by ELISA;Flow cytometry was used to detect the secretion of cytokines in T cells;The brucellosis EipB protein is composed of 279 amino acids,is weakly basic,has a molecular mass of 31 kDa and is relatively stable;contains a signal peptide sequence and a transmembrane structural domain;has a secondary structure with a-helix,β-turn,extended linkage and irregular coi-ling;has a distinct hydrophilic region,and the protein contains 21 dominant B-cell epitope peptides in amino acid regions,the amino acid regions in which these dominant B-cell epitopes are located correspond to regions of higher hydrophilicity.Contains 56 CD4+T cell epitopes and 39 CD8+T cell epitopes.After immunizing mice with T-B dominant cell epitope combination,antibodies such as IgG,IgM,IgA and IgE in serum increased significantly;T cells produced IFN-γ and other cytokines significantly increased.1)Bioinformatics method was used to predict that EipB pro-tein contains dominant B and T cell epitopes.2)Combined immunization of mice with dominant B-T dominant epitope peptides can induce a higher level of immune response in mice,providing a reference for the screening of brucellosis molecular peptide vaccines.

ObjectiveTo investigate the effect of myosin heavy chain 7 gene-derived miRNA-208b-3p on the fibrotic phenotype of cardiac fibroblasts. MethodsmiRNA chip array was performed to detect the dysregulated miRNAs in the myocardium of diabetic db/db mice and db/m control mice. Neonatal mouse ventricular cardiomyocytes （NMVCs） and cardiac fibroblasts （CFs） were isolated from C57BL/6 mice and cultured. Real-time quantitative PCR （RT-qPCR） was conducted to determine the expression of miR-208b-3p in mouse CFs and NMVCs subjected to angiotensinⅡ（AngⅡ） and high glucose plus glucose oxidase （G/Go） treatment， respectively. Cell counting kit 8（CCk8） assay， flow cytometry and determination of fibrosis-related protein， including COL1A1， COL3A1and α-SMA， were performed in mCFs transfected with miR-208b-3p. Dual luciferase reporter assay was performed to confirm the interaction between miR-208b-3p and the 3'-UTR of metal response element binding transcription factor 2 （Mtf2） and progesterone receptor membrane component 1（Pgrmc1）， respectively. The expressions of Mtf2 and Pgrmc1 at the mRNA and protein levels in mCFs after miR-208b-3p mimic transfection were determined using RT-qPCR and Western blot assay， respectively. The small interfering RNA （siRNA） was used to inhibit Mtf2 and Pgrmc1 expression in mCFs， and the effects of Mtf2 siRNA， Pgrmc1 siRNA and miR-208b-3p on fibrosis-related protein expression in mCFs were investigated. ResultsResults of miRNA chip array and RT-qPCR assay showed that miR-208b-3p was up-regulated in the myocardium of the diabetic db/db mice. miR-208b precursor and the host gene of Myh7 were consistently increased in db/db mice. miR-208b-3p and Myh7 mRNA were expressed in mCFs and NMVCs， but the levels of miR-208b-3p and Myh7 mRNA in NMVCs were much higher than those in mCFs. miR-208b-3p was up-regulated in mCFs and NMVCs subjected to Ang Ⅱ and G/Go treatment， respectively. miR-208b-3p could significantly enhance fibrosis-related protein， including COL1A1， COL3A1 and α-SMA， in mCFs， without affecting the proliferation activity and cell cycle distribution of mCFs. Dual luciferase reporter assay revealed the interactions of miR-208b-3p with the 3'-UTR of Mtf2 and Pgrmc1. The results of RT-qPCR and Western blotting confirmed that miR-208b-3p inhibited Mtf2 and Pgrmc1 expression at the post- transcriptional level. Transfection with miR-208b-3p mimic， Mtf2 siRNA and Pgrmc1 siRNA could consistently enhance the fibrosis-related protein expression in the cardiac fibroblasts. ConclusionsmiR-208b-3p enhances fibrosis-related gene expression by targeting Mtf2 and Pgrmc1in mCFs.

OBJECTIVE: To assess the efficacy and safety of Guanxin Danshen Dripping Pills (GXDS) in the treatment of depression or anxiety in patients with coronary heart disease (CHD) after percutaneous coronary intervention (PCI). METHODS: From September 2017 to June 2019, 200 CHD patients after PCI with depression and anxiety were included and randomly divided into GXDS (100 cases) and placebo control groups (100 cases) by block randomization and a random number table. Patients in the GXDS and control groups were given GXDS and placebo, respectively, 0.4 g each time, 3 times daily for 12 weeks. The primary outcomes were scores of Patient Health Questionnaire-9 (PHQ-9), Generalized Anxiety Scale (GAD-7) and the Seattle Angina Pectoris Scale (SAQ). The secondary outcomes included 12 Health Survey Summary Form (SF-12) scores and the first onset time and incidence of major adverse cardiovascular events (MACEs). Other indices including blood pressure, blood lipids, microcirculation and inflammatory-related indices, etc. were monitored at baseline, week 4, and week 12. RESULTS: In the full analysis set (200 cases), after treatment, the PHQ-9 and GAD-7 scores in the GXDS group were considerably lower than those in the control group (P<0.05). Compared with the baseline, the total PHQ-9 scores of the experimental and control groups decreased by 3.97 and 1.18, respectively. The corrected mean difference between the two groups was -2.78 (95% CI: -3.47, -2.10; P<0.001). The total GAD-7 score in the GXDS group decreased by 3.48% compared with the baseline level, while that of the placebo group decreased by 1.13%. The corrected mean difference between the two groups was -2.35 (95% CI: -2.95, -1.76; P<0.001). The degree of improvement in SAQ score, SF-12 score, endothelin and high-sensitive C-reactive protein levels in the GXDS group were substantially superior than those in the placebo group, and the differences between the two groups were statistically significant (P<0.05). Similar results were obtained in the per protocol population analysis of 177 patients. Three cases of MACES were reported in this study (1 in the GXDS group and 2 in the placebo group), and no serious adverse events occurred. CONCLUSIONS GXDS can significantly alleviate depression and anxiety, relieve symptoms of angina, and improve quality of life in patients with CHD after PCI. (Registration No. ChiCTR1800014291).
Humans ; Percutaneous Coronary Intervention/adverse effects* ; Quality of Life ; Depression ; Coronary Disease/drug therapy* ; Drugs, Chinese Herbal/therapeutic use* ; Angina Pectoris/drug therapy* ; Prognosis ; Anxiety ; Treatment Outcome ; Double-Blind Method

Objective:To study the effect of health management based on the transtheoretical model and stages of change on patients with diabetes mellitus complicated by obesity.Methods:A total of 122 patients with type 2 diabetes mellitus (T2DM) complicated by obesity and admitted to Guangdong Second Provincial General Hospital between January 2019 and June 2020 were recruited to participate in the study. They were divided into two equal groups at random, the observation group and the control group —with 61 patients in each group. Both groups were subjected to individualized health management based on conventional drug treatment. The control group received routine health education and psychological support, while the observation group was subjected to the transtheoretical model and stages of change interventions for six months. The independent-samples t-test was used to compare blood glucose, body weight, and self-efficacy in the two groups after interventions, and the χ 2 test or Fisher′s precise test was used to compare behavior habit compliance rates in the two groups after interventions. Results:FBG with (6.27±0.92) mmol/L and (6.58±0.91) mmol/L, 2hPG with (8.53±1.39) mmol/L and (8.53±1.39) mmol/L, and HbA1c with (6.49±0.91)% and (6.78±0.96)% in the two groups were significantly lower at six months of interventions than those before ( P<0.05), but the differences between the two groups were not statistically significant ( P>0.05). BMI, waist circumference, and body fat rate with (30.46±2.51) kg/m 2, (91.72±10.45) cm, and (34.09±3.84)% of the observation group at six months of interventions were significantly lower than those before interventions ( P<0.05), whereas there were no significant changes in the control group before and after interventions ( P>0.05). The observation group′s BMI and body fat rate were lower than those of the control group ( P<0.05). The General Self-Efficacy Scale (GSES) scores in the two groups were (31.62±2.59) points and (28.75±2.48) points, which were significantly higher than those before interventions ( P<0.05). The observation group′s GSES score was higher than that of the control group ( P<0.05). At six months of interventions, the compliance rates of diet control (92.98% and 77.97%), reasonable exercise (87.72% and 71.19%), adherence to medication (98.25% and 91.53%), quitting smoking and alcohol (94.74% and 81.36%), blood glucose monitoring (91.23% and 94.92%), and regular review (100.00% and 98.31%) were all significantly higher in the two groups than those before interventions ( P<0.05). Additionally, the compliance rates of diet control, reasonable exercise, and quitting smoking and alcohol in the observation group were higher than those of the control group ( P<0.05). Conclusion:The use of the transtheoretical model and stages of change for the self-management intervention of diabetes mellitus patients with obesity is beneficial to improve self-efficacy, behavioral habits, and weight control effectiveness.

Objective To develop an Modified Early Warning Score (MEWS) table suitable for professional characteristics and to predict the occurrence of complications in the percutaneous coronary intervention (PCI). Methods This is a retrospective study. PCI patients in catheter room of Qingdao International Center Hospital from July to December, 2015 and July to December, 2016 were selected as the research object. The former was set as the control group, and the latter as the experimental group, traditional MEWS and modified MEWS were applied in the two groups respectively. The incidence of complications were compared between two groups. Results The incidence rates of cardiac arrest, drop of blood pressure, arrhythmia, slow blood flow or no complex flow, perforation, interlayer were 0.83％(3/362), 0.55％ (2/362), 0.55％ (2/362), was 0.55％ (2/362), 2.21％ (8/362) , 4.42％ (16/362) , and were 2.51％(9/358), 5.31％(19/358), 8.66％(31/358) , 3.07％(11/358) , 2.51％(9/358) , 2.79％(10/358) in the control group, the differences were statistically significant (χ2= 4.603-5.302, P<0.05). Conclusions The MEWS score of PCI can be used to predict the occurrence of complications in PCI, and for the patients with MEWS score above 5, they should be given medical and nursing intervention in order to reduce the complications in the operation.

Locally advanced pancreatic cancer is associated with a very poor prognosis. This study was performed to evaluate whether patients with locally advanced pancreatic cancer benefit from (125)I seed implantation. This retrospective study included 224 patients with locally advanced pancreatic cancer, with 137 patients (61.2%) in the implantation (IP) group and 87 (38.9%) in the non-implantation (NIP) group. The survival status, complications and objective curative effects were compared between the groups. The average operative time in the IP group was significantly longer than that in the NIP group (243±51 vs. 214±77 min). The tumor response rates were 9.5% and 0 at the 2nd month after surgery in the IP and NIP groups, respectively (P<0.05). The IP group exhibited a trend toward pain relief at the 6th month after surgery. The global health status scores of the IP group were higher than those of the NIP group at the 3rd and 6th month after surgery. The median survival time in the IP group was significantly longer than that in the NIP group. In conclusion, patients with locally advanced pancreatic cancer can benefit from (125)I seed implantation in terms of local tumor control, survival time, pain relief and quality of life.
Adult ; Aged ; Drug-Eluting Stents ; adverse effects ; Female ; Humans ; Iodine Radioisotopes ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Pancreatic Neoplasms ; pathology ; radiotherapy ; Quality of Life ; Radiopharmaceuticals ; administration & dosage ; therapeutic use ; Survival Analysis

The purpose of this study was to investigate the etiology, pathological characteristics, management and prognosis of chronic pancreatitis in the Chinese population. The clinical data of 142 patients with chronic pancreatitis were retrospectively studied. All patients were of Chinese nationality and hospitalized from January 2008 to December 2011. Their ages ranged from 14 to 76 years, with a mean of 43 years. Of 142 patients, there were 72 cases of obstructive chronic pancreatitis (50.70%), 19 cases of alcoholic chronic pancreatitis (13.38%), 14 cases of autoimmune pancreatitis (9.86%) and 37 cases of undetermined etiology (26.06%). Pathologically, the average inflammatory mass diameter was 3.8 ± 3.3 cm, biliary obstruction occurred in 36 cases, gall stones in 70 cases, calcification in 88 cases, ductal dilatation in 61 cases, side branch dilatation in 32 cases, ductal irregularity in 10 cases, lymphocytic inflammation in 23 cases, obliterative phlebitis in 14 cases, extra pancreatic lesion in 19 cases and fibrosis in 142 cases. Location of pancreatic lesion in the region of head (n=97), neck (n=16), body (n=12), tail (n=15) and whole pancreas (n=2) influenced the choice of surgical procedures. Ninety-four patients (66.20%) received surgical treatment and 33.80% received other treatments. After operation, 80.85% of 94 patients experienced decreased pain, and 8.51% of 94 showed recovery of endocrine function but with a complication rate of 12.77%. All the operations were performed successfully. According to the pain scale of European Organization for Research and Treatment of Cancer (QLQ-C30) a decrease from 76 ± 22 to 14 ± 18 was observed. Etiology, pathological characteristics, management and prognosis of chronic pancreatitis in the Chinese population vary from others.
Adolescent ; Adult ; Aged ; Autoimmune Diseases ; epidemiology ; therapy ; China ; epidemiology ; Cholestasis ; epidemiology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Pancreatitis, Alcoholic ; epidemiology ; therapy ; Pancreatitis, Chronic ; etiology ; pathology ; therapy ; Prognosis ; Retrospective Studies ; Treatment Outcome ; Young Adult

The main treatment strategies for chronic pancreatitis in young patients include therapeutic endoscopic retrograde cholangio-pancreatography (ERCP) intervention and surgical intervention. Therapeutic ERCP intervention is performed much more extensively for its minimally invasive nature, but a part of patients are referred to surgery at last. Historical and follow-up data of 21 young patients with chronic pancreatitis undergoing duodenum-preserving total pancreatic head resection were analyzed to evaluate the outcomes of therapeutic ERCP intervention and surgical intervention in this study. The surgical complications of repeated therapeutic ERCP intervention and surgical intervention were 38% and 19% respectively. During the first therapeutic ERCP intervention to surgical intervention, 2 patients developed diabetes, 5 patients developed steatorrhea, and 5 patients developed pancreatic type B pain. During the follow-up of surgical intervention, 1 new case of diabetes occurred, 1 case of steatorrhea recovered, and 4 cases of pancreatic type B pain were completely relieved. In a part of young patients with chronic pancreatitis, surgical intervention was more effective than therapeutic ERCP intervention on delaying the progression of the disease and relieving the symptoms.
Adolescent ; Adult ; Cholangiopancreatography, Endoscopic Retrograde ; adverse effects ; methods ; Diabetes Mellitus ; etiology ; Female ; Follow-Up Studies ; Humans ; Male ; Pain, Postoperative ; etiology ; Pancreatectomy ; adverse effects ; methods ; Pancreatitis, Chronic ; surgery ; Postoperative Complications ; etiology ; Steatorrhea ; etiology ; Treatment Outcome ; Young Adult