To reduce the dependency on high-carbon-load chromatographic columns,a new method has been established for the determination of the content of docusate sodium using ion-pair high-performance liquid chromatography (IP-HPLC). Tetrapropylammonium chloride was used as the ion-pair reagent with a mobile phase, composition of acetonitrile:10 mmol/L tetrapropylammonium chloride solution = 66∶34, adjusting pH to 6.5 with 0.1% phosphoric acid solution,flow rate of 1.5 mL/min, detection wavelength of 214 nm,column temperature of 35 °C, and an injection volume of 25 μL,and quantified by an external standard method. The main peak of docusate sodium exhibited a tailing factor of 1.34. The method showed good linearity within the range of 0.02 mg/mL to 0.40 mg/mL, with a correlation coefficient (r) of 0.999 9. It also demonstrated good repeatability, with recovery ranging from 97.0% to 98.2% (n=6). The quantification limit was 3.31 μg/mL, and the detection limit was 2.76 μg/mL.In summary,the new method shows good durability, a wide linear range, and high sensitivity, it is suitable for the determination of docusate sodium.

OBJECTIVE: To elucidate the role and underlying mechanism of ubiquitin-specific protease 35 (USP35) in ferroptosis of rheumatoid arthritis-fibroblast like synoviocytes (RA-FLS), thereby enhancing our comprehension of the pathogenesis of RA and identifying potential therapeutic targets for its treatment. METHODS: (1) RA-FLS were cultured in vitro and transduced with lentiviral vectors to establish stable cell lines: A USP35-knockdown line (short hairpin ribonucleic acid of USP35, shUSP35) and its control (negtive control of short hairpin ribonucleic acid, shNC), as well as a overexpression of USP35 line (USP35 OE) and its control (Vector). To investigate the role of USP35 in ferroptosis regulation, a ferroptosis model was induced in RA-FLS by treatment with 1 μmol/L Erastin. The cells were divided into six groups: shNC, shNC + Erastin, shUSP35 + Erastin, Vector, Vector + Erastin, and USP35 OE + Erastin. (2) Cell viability was detected using the cell counting kit-8 (CCK-8). (3) Reactive oxygen species (ROS), malondialdehyde (MDA), glutathione/glutathione disulfide (GSH/GSSG) ratios, and Ferrous ion (Fe²⁺) levels were measured using specific assay kits to evaluate oxidative stress, lipid peroxidation, and glutathione redox status in the cells. (4) Protein expression levels of solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) were detected using Western blotting to investigate their potential involvement in USP35-mediated ferroptosis regulation. RESULTS: (1) Compared with the shNC +Erastin group, the cell viability of the shUSP35+Erastin group was significantly decreased (P < 0.001), while it was notably increased in the USP35 OE+Erastin group compared with the Vector+Erastin group (P < 0.001). These findings indicated that USP35 could alleviate the inhibitory effect of Erastin on RA-FLS cell viability. (2) In comparison to the shNC+Erastin group, the levels of ROS (P < 0.001), MDA (P < 0.05), and Fe²⁺ (P < 0.001) were significantly elevated, and the GSH/GSSG ratio was increased (P < 0.05) in the shUSP35+Erastin group. Conversely, the levels of ROS (P < 0.001), MDA (P < 0.05), and Fe²⁺ (P < 0.05) were significantly decreased, and the GSH/GSSG ratio was decreased (P < 0.05) in the USP35 OE+Erastin group compared with the Vector+Erastin group. These results suggested that USP35 could inhibit Erastin-induced oxidative stress and lipid peroxidation in RA-FLS. (3) In Erastin-induced RA-FLS, the expression of USP35 was positively correlated with the protein levels of SLC7A11 and GPX4, indicating a potential mechanism by which USP35 regulated ferroptosis in these cells. CONCLUSION USP35 inhibits ferroptosis in RA-FLS, potentially through the increased expression of SLC7A11 and GPX4.
Ferroptosis ; Humans ; Arthritis, Rheumatoid/metabolism* ; Synoviocytes/pathology* ; Reactive Oxygen Species/metabolism* ; Ubiquitin-Specific Proteases/metabolism* ; Fibroblasts/pathology* ; Cell Survival ; Piperazines/pharmacology* ; Endopeptidases/metabolism* ; Cells, Cultured ; Cell Line ; Amino Acid Transport System y+

To reduce the dependency on high-carbon-load chromatographic columns,a new method has been established for the determination of the content of docusate sodium using ion-pair high-performance liquid chromatography(IP-HPLC).Tetrapropylammonium chloride was used as the ion-pair reagent with a mobile phase,composition of acetonitrile:10 mmol/L tetrapropylammonium chloride solution=66∶34,adjusting pH to 6.5 with 0.1%phosphoric acid solution,flow rate of 1.5 mL/min,detection wavelength of 214 nm,column temperature of 35℃,and an injection volume of 25 μL,and quantified by an external standard method.The main peak of docusate sodium exhibited a tailing factor of 1.34.The method showed good linearity within the range of 0.02 mg/mL to 0.40 mg/mL,with a correlation coefficient(r)of 0.999 9.It also demonstrated good repeatability,with recovery ranging from 97.0%to 98.2%(n=6).The quantification limit was 3.31 μg/mL,and the detection limit was 2.76 μg/mL.In summary,the new method shows good durability,a wide linear range,and high sensitivity,it is suitable for the determination of docusate sodium.

Objective:To investigate the efficacy and safety of high semi-decubitus position in the second stage of labor in epidural analgesic delivery of primipara.Methods:A total of 120 women who gave birth in Hang-zhou women's Hospital from May 1,2021 to July 30,2021 were randomly divided into experimental group(n=60)and control group(n=60)by random number table method.The control group adopted supine position in the sec-ond stage of labor,and the experimental group transferred to high semi-supine position after the supine position was used applied for 30 minutes when the fetal head was not exposed.To compare the effects of different postur-al positions on the duration of the second stage of labor,mode of delivery,postpartum injury,perineal tearing,la-bor force experience and labor control,and neonatal asphyxia.Results:The duration of uterine opening to fetal head exposure,the duration of uterine opening to fetal head crown and the duration of second stage of labor in experimental group were shorter than those in control group.The rate of natural delivery of experimental group was higher than that of control group.The episiotomy rate and the second degree perineal laceration rate were lower than that of control group.The amount of blood loss during delivery and 2h postpartum of experimental group was less than that of control group.The scores of birth experience and birth control of experimental group were higher than those of control group,and the differences were statistically significant(P＜0.05).There was no significant difference in 1-minute Agpar score and 5-minute Agpar score between the two groups(P＞0.05).Conclusions:The application of high semi-decubitus position under epidural labor analgesia promoted vaginal natural labor,shortened the time of second stage of labor,reduced episiotomy rate and perineal laceration de-gree,reduced postpartum hemorrhage,had positive labor force experience,and increased the sense of labor con-trol.

Objective:To assess the efficiency of modified enrichment method for cell-free fetal DNA (cffDNA) through purified superparamagnetic beads during non-invasive prenatal testing (NIPT).Methods:A total of 26 252 pregnant women undergoing NIPT at the Maternal and Child Health Care Hospital of Haidian District from December 2017 to September 2022 were recruited and randomly assigned into the conventional group ( n = 10 573) and the modified enrichment group ( n = 15 679), who were then subjected to the screening and enrichment of the cffDNA using a conventional and modified technique, respectively. High-risk pregnant women detected by NIPT were subjected to invasive prenatal diagnosis. All women were followed up for their pregnancy outcomes, and the detection efficacy of the two methods was compared in terms of fragment size, concentration of cffDNA, duplicate detection rate, and indices of clinical laboratory tests. Results:The fragment size of the main peak of the cell-free DNA library of the modified enrichment group was significantly lower than that of the conventional group [267 (264, 269) bp vs. 294 (292, 296) bp, P<0.01], while the concentration of cffDNA was significantly higher [21.86% (17.61%, 26.36%) vs. 9.08% (6.87%, 11.87%), P<0.01]. In addition, the duplicate detection rate (0.740% vs. 2.02%, χ2=83.90, P<0.01) and detection failure rate (0.006% vs. 0.057%, P<0.05) in the modified enrichment group were significantly lower than those of the conventional group. The combined positive predictive value (PPV) in both high-risk (64.3% vs. 76.1%) and low-risk (35.3% vs. 45.5%) pregnant women from the modified enrichment group was slightly lower than those from the conventional group, though no significant difference was detected. There was one false negative case for trisomy 21 among the high-risk pregnant women from the conventional group, and no false negative case was found in the modified enrichment group. Conclusion:The modified technique to screen and enrich the cffDNA has significantly enhanced the relative concentration of cffDNA and reduced the failure and duplication detection rate of NIPT, which has significantly reduced the incidence of false negative cases due to the low concentration of cffDNA, and greatly increased the overall detection efficacy of NIPT.

Objective:To study the clinical features and genotypes of neonatal Glanzmann thrombasthenia(NGT).Methods:A male neonate with NGT admitted to the Department of Neonatology of our hospital was retrospectively reviewed. CNKI, Wangfang database, VIP, the Chinese Medical Journal Full Text database, PubMed and Embase database were searched using key words '(neonate OR newborn) AND (Glanzmann thrombasthenia)' both in English and Chinese. The clinical features and genotypes of NGT were summarized and analyzed.Results:A male full-term neonate was admitted to our hospital for mass on the forehead and ecchymosis and petechiae on the body within half an hour after birth. He gradually developed subgaleal hemorrhage and severe anemia. Platelet count, mean platelet volume and coagulation functions were normal. The platelet aggregation test indicated decreased platelet aggregation rate induced by arachidonic acid and adenosine diphosphate. Genetic testing revealed two heterozygous mutations in the patient's ITGA2B gene: NM_000419.4: c.886G>A(p.Gly296Arg) and NM_000419.4: c.2855dup(p.Phe953Valfs*83). A total of 42 literature involving 44 patients (our case included) with NGT were retrieved. 33 cases (75.0%) of NGT showed ecchymosis or petechiae on the first day after birth. For 13 cases with detailed information, 5 cases with severe anemia were given erythrocyte and plasma transfusion and platelet transfusion was given in 1 case. 4 cases had homozygous variants and 4 cases showed compound heterozygous variants. 10 cases had follow-up records, including 2 cases without any bleeding and 8 cases with varying degrees of bleeding during follow-up. No deaths were reported.Conclusions:Neonates with ecchymosis and petechiae in the early postnatal period should be suspected of NGT. Blood transfusion is preferred when the indication for transfusion is met.

Objective:To evaluate the value of modified magnetic bead screening for enrichment of cell-free fetal DNA (cffDNA) in non-invasive prenatal testing (NIPT).Methods:This study retrospectively recruited 31 cases with low concentration of cffDNA (<6.00%), Z value in the gray zone (3.00-4.00) at the first detection, or false-positive (confirmed by invasive prenatal diagnosis) or false-negative (confirmed by postnatal chromosome test) results among 11 000 pregnant women who underwent routine NIPT in Beijing Haidian District Maternal and Child Health Care Hospital from October 2017 to December 2019. Plasma samples collected for the first-time routine NIPT were used to enrich cffDNA using modified magnetic beads for NIPT (modified NIPT). Wilcoxon rank sum test was used to compare the modified NIPT with the routine NIPT in detecting the cffDNA concentrations of male fetuses.Results:Among the 31 pregnant women, there were 13 cases with low cffDNA concentration in routine NIPT, 11 having false-positive results in the routine NIPT (three for trisomy 13, four for trisomy 18 and four for trisomy 21, all were confirmed by invasive prenatal diagnosis), six with gray-zone Z values in the first-time NIPT (retesting indicating low risk) and one having false negative result for trisomy 21 (confirmed by postnatal chromosome test). Cell-free DNA (cfDNA) fragments less than 150 bp were effectively enriched using the modified magnetic bead screening and the concentration of cffDNA was increased from 4.43% (2.45%-17.61%) in routine NIPT to 13.46% (7.75%-36.64%) in the modified NIPT ( Z=-14.22, P<0.01). Results of the modified NIPT indicated that 13 cases with low cffDNA concentration of routine NIPT were successfully detected as low risk, as well as the risks in the six cases with gray-zone Z value and six of the 11 false-positive cases in the routine NIPT were low, which were consistent with the retest results of the routine NIPT, while high risk was found in one false-negative case. Conclusions:The modified NIPT could reduce the false positive rate by lowering the failure rate caused by low concentration of cffDNA and is able to identify false-negative cases. Compared with the routine NIPT, it shows a higher success rate and a lower false positive rate.

Objective:To investigate the clinical effect of upper arm central venous port in cancer patients.Methods:A total of 500 patients with tumor were selected as the study subjects from March 29, 2018 to January 19 2020. Complications such as catheter-related thrombosis and catheter-related bloodstream infection during the indwelling period were recorded.Results:One patient with severe superior vena cava syndrome failed to be intubated. The other 499 patients were successfully intubated with a success rate of 99.8% (499/500). The rate of complications was 2.6% (13/499), and the rate of catheter-related bloodstream infection was 0.029‰ (4/139 614) in 4 cases, the incidence of catheter-related thrombus was 0.057‰ (8/139 614) in 8 cases. There were 1 case of body turnover, 1 case of median nerve injury, 1 case of local tissue necrosis and 1 case of lymphatic leakage. Extubation was planned in 15 patients, and unplanned extubation in 6 patients due to complications.Conclusions:The upper arm port has less complications and is suitable for patients with tumor chemotherapy and long-term need of infusion. It is worthy of clinical promotion.

Objective:To investigate the clinical efficacy of pancreaticodudenectomy (PD) using anterior approach in situ technique for pancreatic head cancer.Methods:The retrospective cohort study was conducted. The clinicopathological data of 285 patients with pancreatic head cancer who were admitted to the First Affiliated Hospital of Kunming Medical University from January 2012 to June 2018 were collected. There were 164 males and 121 females, aged from 40 to 76 years, with an average age of 57 years. Of the 285 patients, 196 patients who underwent PD using anterior approach in situ technique were set as anterior approach group, 89 patients who underwent PD using traditional approach were set as traditional approach group. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow-up was performed by outpatient examination, telephone or network interview once every 2 to 3 months to detect tumor recurrence, metastasis and survival of patients up to December 2018. The endpoint of follow-up was death of patients, and the secondary endpoint of follow-up was tumor recurrence or metastasis. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed by the t test. Measurement data with skewed distribution were represented as M (range), and comparison between groups was analyzed by the Mann-Whitney U test. Count data were represented as absolute numbers or percentages, and comparison between groups was analyzed by the chi-square test. Kaplan-Meier method was used to draw the survial curve and calculate the survival rates. Survival analysis was done by the Log-rank test. Results:(1) Surgical situations: 285 patients underwent surgery successfully. Cases with pylorus-preservaction, cases with superior mesenteric vein/ portal vein (SMV/PV) resection and reconstruction (end to end anastomosis, artificial vascular replacement, lateral wall resection and anastomosis), operation time, volume of intraoperative blood loss were 118, 37 (17, 11, 9), (303±107)minutes, 350 mL(range, 100-750 mL) in the anterior approach group, and 48, 9 (7, 1, 1), (335±103)minutes, 400 mL(range, 100-900 mL) in the traditional approach group, respectively, showing no significant difference between the two groups ( χ2=0.990, 3.474, t=0.722, Z=1.729, P>0.05). (2) Postoperative situations: the rate of R 0 resection, the number of lymph node dissected, the number of positive lymph node dissected, rate of nerve invasion, rate of vascular invasion, cases with postoperative severe complica-tions, the number of perioperative death, cases with postoperative chemotherapy were 93.88%(184/196), 12(range, 5-19), 4(range, 0-15), 45.41%(89/196), 31.12%(61/196), 28, 3, 69 in the anterior approach group, and 85.39%(76/89), 7(range, 4-17), 5(range, 0-13), 32.58%(29/89), 23.60%(21/89), 11, 2, 41 in the traditional approach group, respectively. There were significant differences in the rate of R 0 resection, the number of lymph node dissected, rate of nerve invasion between the two groups ( χ2=5.506, Z=4.637, χ2=4.149, P<0.05), while there was no significant difference in the number of positive lymph node dissected, rate of vascular invasion, cases with postoperative severe complications, the number of perioperative death, cases with postoperative chemotherapy between the two groups ( Z=0.052, χ2=1.962, 0.192, 0.001, 3.048, P>0.05). (3) Follow-up: of the 285 patients, 252 and 228 achieved the secondary endpoint and the endpoint of follow-up respectively with the follow-up time of 35 months (range, 6-58 months). There were 181 and 176 of 196 patients in the anterior approach group achieving the secondary endpoint and the endpoint of follow-up respectively with the follow-up time of 38 months (range, 6-58 months). There were 71 and 52 of 89 patients in the traditional approach group achieving the second endpoint and the endpoint of follow-up respectively with the follow-up time of 33 months (range, 7-53 months). The median tumor free survival time and median overall survival time were 31 months and 37 months in the anterior approach group, respectively, versus 24 months and 31 months in the traditional approach group. There was a significant difference in the tumor free survival between the two groups ( χ2=7.646, P<0.05), while no significant difference in the overall survival between the two groups ( χ2=3.265, P>0.05). Conclusion:PD using anterior approach in situ technique is safe and feasible for pancreatic head cancer, which can improve the rate of R 0 resection and prolong the tumor free survival time of patients.

Objective: To screen for hotspot gene mutations associated with genetic deafness in Chinese pregnant women, and to perform risk assessment and prenatal diagnosis in high-risk families. Methods: Between November 2012 and October 2017, 26 117 pregnant women were screened by molecular hybridization microarray for 9 hot-spot mutations in 4 hereditary deafness related genes (GJB2 c. 35 del G, c. 176_191 del 16 bp, c. 235 del G, c. 299_300 del AT, GJB3 c. 538 C>T, SLC26A4 c. 2168 A>G, IVS 7-2 A>G, mitochondrial DNA 12S rRNA m. 1494 C>T, m. 1555 A>G). Genotype analysis was carried out in husbands of women carrying mutations, and prenatal diagnosis was carried out in the fetuses with high risk of deafness. Results: Among all women tested, 1 208(4.63%) were carriers of genetic deafness mutations, 7 with hearing impairment were affected by homozygous or compound heterozygous mutations, 51 were mitochondrial gene mutation carriers, 103 were carriers of GJB3 c. 538 C>T heterozygous mutation, 1 026 were carriers of GJB2 or SLC26A4 heterozygous mutations, and 21 carried heterozygous mutations in two genes simultaneously. In 394 families, the husbands accepted gene sequence testing, and 27 in which were determined as carriers of mutations in identical genes as their wives. Among which, 18 families received prenatal diagnosis, and 5 fetuses were diagnosed as hereditary deafness. In 9 families who did not receive prenatal diagnosis, 1 neonate was diagnosed as compound heterozygote after delivery. Conclusion In order to prevent birth defects with congenital hearing problems, it is effective to provide screening for hotspot mutations in pregnant women and to perform prenatal diagnosis on high risk pregnancies.