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.

BACKGROUND:The development of steroid-induced osteonecrosis of the femoral head is a complex process involving multiple mechanisms.There is still no standard therapeutic drug for early intervention of this disease.Current studies have shown that baicalein has various pharmacological activities such as regulating lipid metabolism,bone metabolism,apoptosis and anti-oxidative stress,which provides an idea for the prevention and treatment of steroid-induced osteonecrosis of the femoral head. OBJECTIVE:To observe the preventive effect of baicalein against steroid-induced osteonecrosis of the femoral head and to investigate its possible mechanism. METHODS:Thirty-six 10-week-old male Sprague-Dawley rats were randomly divided into three groups(n=12 per group):blank control group,model group,and baicalein intervention group.In the model group and baicalein intervention group,intraperitoneal lipopolysaccharide and intramuscular injection of methylprednisolone sodium succinate were performed for modeling,while normal saline was used as a substitute for the modeling drug in the blank control group.Baicalein 300 mg/kg was administered by gavage(once a day for 6 weeks)at the time of initial intramuscular glucocorticoid injection in the baicalein intervention group,and baicalein was replaced by normal saline in the other two groups.The serum level of malondialdehyde in rats was detected at 2 weeks of the experiment.Blood lipid indicators and bone formation metabolic markers were detected at 6 weeks of the experiment,the histomorphometric changes of the femoral head were analyzed by hematoxylin-eosin staining,anti-tartaric acid phosphatase staining and TUNEL staining,and the femoral head was subjected to Micro-CT scanning and three-dimensional reconstruction of the bone in order to analyze the alterations of bone tissue structure and parameters. RESULTS AND CONCLUSION:The serum levels of malondialdehyde,triglyceride,β-collagen type Ⅰ carboxy-terminal peptide were increased and the serum levels of bone specific alkaline phosphatase and pre-collagen type Ⅰ amino-terminal peptide were decreased in the model group compared with the blank control group(P＜0.05).The serum level of malondialdehyde decreased in the baicalein intervention group compared with the model group(P＜0.05),but there was no significant difference between the baicalein intervention group and blank control group(P＞0.05).The serum level of triglyceride was higher in the baicalein intervention group than the blank control group(P＜0.05),but had no significant difference between the baicalein intervention group and model group(P＞0.05).There were also no significant differences in the levels of bone specific alkaline phosphatase and β-collagen type Ⅰ carboxy-terminal peptide between the baicalein intervention group and the other two groups(P＞0.05).The serum level of the baicalein intervention group was lower in the baicalein intervention group than the blank control group(P＜0.05)but had no significant difference between the baicalein intervention group and model group(P＞0.05).Histomorphological analysis of the femoral head showed that the rate of bone empty lacuna,osteoclast counting and cell apoptosis rate in the femoral head of model group rats were significantly higher than those of the other two groups(P＜0.05).There was a significant increase in the number of adipocytes in the bone marrow cavity of the femoral head,bone trabeculae were thinned and sparsely arranged with more disruptions in the continuity.The incidence of osteonecrosis was higher in the model group(75%)than in the baicalein intervention group(25%;bilateral and unilateral exact significance results were both 0.05).There was also an increase in the number of adipocytes in the bone marrow cavity of the femoral head in the baicalein intervention group,and the trabecular changes were roughly similar to those in the model group.Micro-CT results showed that bone volume fraction,trabecular thickness,trabecular number,and bone mineral density decreased and trabecular separation increased in the model group compared with the blank control group(P＜0.05).Overall significant bone mass loss was observed in the model group.Bone tissue parameters in the baicalein intervention group were significantly improved than those in the model group,which were reflected in bone volume fraction,trabecular thickness and trabecular separation(P＜0.05),and trabecular number and bone mineral density had no significant difference between the baicalein intervention group and blank control group(P＞0.05).Although baicalein failed to significantly ameliorate dyslipidemia and promote bone formation in rats with steroid-induced osteonecrosis of the femoral head,it could reduce the incidence of steroid-induced osteonecrosis of the femoral head by reducing oxidative stress damage,decreasing cell apoptosis,inhibiting osteoclasts,suggesting its effectiveness in the early prevention of steroid-induced osteonecrosis of the femoral head.

Objective:To evaluate the risk factors for the formation of parastomal Hernias(PSH)using meta-analysis,and to provide a theoretical basis for the prevention and treatment of PSH.Methods:Case control or Cohort study of PSH risk factors were collected by searching PubMed,CNKI,Wanfang data and other databases.Extract relevant data and perform meta-analysis using RevMan 5.3.Results:The results included a total of 16 studies,with a total sample size of 2411 cases,including 670 in the PSH group and 1741 in the non PSH group.The results showed that advanced age,female gender,BMI≥25,hypertension,COPD/chronic cough,diabetes,and postoperative Hypoproteinemia could increase the risk of PSH(P＜0.05);Smoking,previous ab-dominal surgery history,preoperative radiotherapy/chemotherapy etc.,were not significantly asso-ciated with the occurrence of PSH(P＞0.05).Conclusion:The current evidence shows that ad-vanced age,female gender,BMI≥25,hypertension,COPD/chronic cough,diabetes,postoperative Hypoproteinemia are risk factors for PSH,and extraperitoneal stoma can reduce the occurrence of PSH.

AIM: To compare the differences and agreement of anterior segment biometric parameters of myopic patients measured by domestic Scansys and the imported Sirius based on the principle of Scheimpflug imaging technique.METHODS: In this case series study, 103 cases(103 eyes)that underwent pre-refractive surgery(including small incision lenticule extraction, femtosecond laser-assisted in situ keratomileusis, transepithelial photorefractive keratectomy and implantable contact lens implantation)at Aier Excellent Eye Hospital from May 2022 to October 2022 were recruited. Preoperative keratometry(Km), central corneal thickness(CCT), anterior chamber depth(ACDEndo.), anterior chamber angle(ACA), anterior chamber volume(ACV), white to white(WTW)of patients were recorded.RESULTS: The results of Km, CCT, ACA, and WTW measured by Scansys and Sirius were 42.88(41.54, 44.60)and 42.98(41.56, 44.52)D,(541.52±29.08)and(549.55±29.62)μm, 42.70°±2.67° and 46.63°±5.13°, 12.10±0.60 and 11.98±0.47 mm, respectively, showing the difference was statistically significant(all P<0.01). The ACV measured by Scansys and Sirius was 194.26±31.06 and 191.47±25.65 mm3, and ACDEndo. was 3.40(3.17, 3.57)and 3.43(3.19, 3.56)mm, with no statistically significant difference(all P>0.05). The range of Km, CCT, ACA, ACDEndo., ACV and WTW values measured by the two instruments was small, with an average difference close to zero, and the points percentage of 95% limits of agreement(LoA)was <5%, which is of good consistency.CONCLUSIONS: Scansys and Sirius have small differences and good agreement in the parameters, which can be replaced by each other in clinical practice. Scansys could theoretically be used to extrapolate the implantable contact lens model or could be a new option for anterior segment parameter measurements.

Objective To investigate the influencing factors and establish a model predicting the performance of needle visualization in fine-needle aspiration (FNA) of thyroid nodules. Methods This study prospectively included 175 patients who underwent FNA of thyroid nodules in the Department of Ultrasound in China-Japan Friendship Hospital and compared the display of the needle tips in the examination of 199 thyroid nodules before and after the application of needle visualization.We recorded the location,the positional relationship with thyroid capsule,ultrasonic characteristics,and the distribution of the soft tissue strip structure at the puncture site of the nodules with unclear needle tips display before using needle visualization.Furthermore,according to the thyroid imaging reporting and data system proposed by the American College of Radiology,we graded the risk of the nodules.Lasso-Logistic regression was employed to screen out the factors influencing the performance of needle visualization and establish a nomogram for prediction. Results The needle tips were not clearly displayed in the examination of 135 (67.8%) and 53 (26.6%) nodules before and after the application of needle visualization,respectively,which showed a significant difference (P<0.001).Based on the positional relationship between the nodule and capsule,anteroposterior/transverse diameter (A/T) ratio,blood supply,and the distribution of subcutaneous strip structure at the puncture site,a nomogram was established to predict the probability of unclear display of the needle tips after application of needle visualization.The C-index of the prediction model was 0.75 (95%CI=0.67-0.84) and the area under the receiver operating characteristic curve was 0.72.The calibration curve confirmed the appreciable reliability of the prediction model,with the C-index of 0.70 in internal validation. Conclusions Needle visualization can improve the display of the needle tip in ultrasound-guided FNA of thyroid nodules.The nomogram established based on ultrasound features such as the positional relationship between the nodule and capsule,A/T ratio,blood supply,and the distribution of subcutaneous strip structure at the puncture site can predict whether needle visualization is suitable for the examination of nodules.
Humans ; Thyroid Nodule/diagnostic imaging* ; Biopsy, Fine-Needle/methods* ; Reproducibility of Results ; Ultrasonography ; Retrospective Studies ; Thyroid Neoplasms

Keratopathy-associated cataract, that is, on the basis of corneal disease, and later the development of lens opacity, seriously damage visual quality. In order to avoid corneal transplantation for some patients, partial visual quality can be restored. A comprehensive and accurate evaluation of the effect of corneal opacity on visual function is of great improtance for determining cataract surgery alone. Due to the opacity of the cornea, the operation is very difficult and challenging. Therefore, it is of clinical value to develop and use new assistive technologies, including capsule staining, endoillumination, pupil dialation technology, femtosecond laser assisted technology, etc., avoiding problems such as limited visibility and decreased light flow caused by corneal opacity and facilitating cataract surgery. This article reviews progress of assistive technologies for keratopathy-associated cataract, hoping to guide clinical application.

AIM: To observe the growth status and morphological changes of primary cultured bulbar conjunctival fibroblasts in different stages of conjunctivochalasis(CCH), and to determine the best passage time, so as to obtain stable and consistent CCH bulbar conjunctival fibroblasts.METHODS: CCH primary bulbar conjunctival fibroblasts were obtained by tissue block adhesion method. The fibroblasts were purified by trypsin differential digestion method. The growth status and morphological changes of fibroblasts in different periods were observed and recorded under inverted microscope. The fibroblasts were identified by immunofluorescence cytochemical staining.RESULTS: After 24h of CCH conjunctival tissue adherent to the wall, a small number of cells would be seen crawling out around the tissue blocks. The logarithmic phase of cell growth was from the 2-7d. The cells grew fast, with vigorously proliferation, clear outline, uniform distribution, increas in numbers and clear nuclei. From the 9-15d, the cell growth entered the plateau stage, the tissue blocks gradually aged and lost activity. The cells grew slowly, arranged loosely, the volume became larger, the shape became flat, and a large number of granular substances and vesicles were produced in the cytoplasm. Some cells fell off from the bottom of the culture bottle, and large gaps appeared between the cells. After subculture and purification, the size and morphology of fibroblasts were basically the same. Through cell identification, fibroblasts were long spindle, flat star or multi-process spindle, wide in the middle, oval nucleus, relatively small at both ends, with 2-3 slender processes of different lengths extending outward.CONCLUSION: Primary CCH bulbar conjunctival fibroblasts can be successfully obtained by tissue block adhesion method. When the cells grow to the 8d, they can be digested and passaged to obtain stable and consistent CCH conjunctival fibroblasts.

ObjectiveTo explore the optimal formula of Maxing Shigantang in regulating epidermal growth factor receptor（EGFR）expression and alleviating airway injury in asthmatic rats and to reveal the underlying mechanism. MethodSD male rats were randomly divided into normal group, model group, dexamethasone group (5×10^-4 g·kg^-1) and Maxing Shigantang 1∶0.5, 1∶1, 1∶2 groups (group A, B, C, 10 g·kg^-1), with 8 rats in each group. The other groups except the normal group received nebulization of 2% acetylcholine chloride and 0.4% histamine phosphate for the modeling of asthma. One hour before modeling， the normal group and the model group were given the same amount of normal saline， and the other groups were given the same amount of corresponding drugs， once a day for 7 days. On the 7^th day， the model was established and the incubation period of asthma was recorded. The rats were then immediately anesthetized， and arterial blood and tracheal tissue were collected. Enzyme-linked immunosorbent assay （ELISA） was employed to detect the levels of interleukin-2 （IL-2）， interleukin-4 （IL-4）， and tumor necrosis factor-α （TNF-α） in serum. Pathological sections were prepared for the observation of the pathological changes of tracheal tissues and the ultrastructure of epithelial cells in each group. Terminal-deoxynucleotidyl transferase-mediated nick-end labeling （TUNEL） was adopted to detect epithelial cell apoptosis， and in situ hybridization and Western blot were employed to determine the mRNA and protein levels of epidermal growth factor receptor （EGFR）， respectively. ResultCompared with the model group， groups A， B and C prolonged the incubation period of asthma （P<0.05，P<0.01）. Compared with the control group， the model group showed declined IL-2 level （P<0.01）， risen IL-4 and TNF-α levels （P<0.05，P<0.01）， increased airway pathology score， collagen volume fraction， and airway epithelial cell apoptosis index （P<0.01）， and up-regulated mRNA and protein levels of EGFR in trachea tissue （P<0.01）. Compared with the model group， group A showed increased IL-2 level （P<0.05） and declined IL-4 （P<0.05，P<0.01） level， and group B showed declined IL-4 level （P<0.05）. The level of TNF-α in groups A， B， and C declined compared with that in the model group （P<0.01）. Maxing Shigantang repaired the tracheal tissue to different degrees （P<0.05）. Among the three groups， group A inhibited tracheal fibrosis （P<0.05） and had the most significant effect of repairing the ultrastructural changes of airway epithelial cells. Groups A， B and C all inhibited the apoptosis of airway epithelial cells （P<0.05）. All the three groups inhibited the up-regulation of EGFR mRNA level （P<0.05，P<0.01）， and groups B and C inhibited the up-regulation of EGFR protein level （P<0.05，P<0.01）. ConclusionMaxing Shigantang can inhibit the abnormal changes of airway epithelial structure， alleviate airway injury， and can down-regulate the expression of EGFR in the tracheal tissue of asthma model rats. In this study， the optimal compatibility of Maxing Shigantang to repair airway epithelial injury in asthmatic rats was group A， with the Ephedrae Herba-Armeniacae Semen Amarum-Glycyrrhizae Radix et Rhizoma-Gypsum Fibrosum ratio of 1∶0.5∶4∶1.

Objective: To explore the knowledge, attitude and behavior of salt reduction in adults of Beijing in 2017. Methods: Based on the monitoring data of chronic diseases and corresponding risk factors in adults of Beijing in 2017, the indicators of salt reduction knowledge, attitude and behavior of 13 240 participants aged 18-79 years old were analyzed. The awareness rate, attitude support rate and behavior rate were calculated by complex weighting method, and compared among different age groups, genders, residential areas, and history of hypertension. The proportion of people taking various salt reduction measures to the total number of people was compared. Results: The awareness rate of recommended daily salt intake, the awareness of hypertension caused or aggravated by more salt intake, the attitude support rate and behavior rate of adults were 31.77%, 88.56%, 90.27% and 53.86%, respectively. After weighted adjustment, the awareness rate of recommended daily salt intake was 31.08%, which increased with age (χ²_trend=431.56, P<0.001) and education level (χ²_trend=95.44, P<0.001). The awareness rate of women was higher than that of men (χ²=118.89, P<0.001), and the awareness rate of population in urban areas was higher than that of population in suburban areas (χ²=34.09, P=0.001). The awareness rate of hypertension caused or aggravated by eating more salt was 86.73%. The support rate of salt reduction attitude was 90.45%. The rate of salt-reducing behavior was 54.05%. Among different salt reduction measures, reducing salt when cooking was the most common measure (52.41%), while the least common one (35.22%) was using low sodium salt. Logistic regression model analysis showed that the gender, age, education level, self-reported history of hypertension, awareness of salt recommendation, awareness of hypertension caused or aggravated by eating more salt, and salt reduction attitude were significantly associated with salt reduction behavior. Conclusion: In 2017, adults in Beijing have a basic understanding of the impact of high-salt diet on health and support salt reduction, but the rate of salt reduction behavior is still relatively low. There are obvious gender and age differences, and the salt reduction measure is simple. Targeted measures should be taken to promote the formation of salt reduction behavior.
Adolescent ; Adult ; Aged ; Beijing ; Diet, Sodium-Restricted ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Hypertension/prevention & control* ; Male ; Middle Aged ; Recommended Dietary Allowances ; Sodium Chloride, Dietary ; Young Adult

Objective: To understand the regular exercise status and influencing factors in 18-64 year-old labor force population in Beijing, and provide evidences for behavioral intervention. Methods: Data were collected from Beijing Adult Non-communicable and Chronic Diseases and Risk Factors Surveillance Program from August to December, 2017. The stratified cluster sampling method was used to select 13 240 survey subjects throughout the city. The valid sample size included in this study were 11 604 persons aged 18 to 64 years. Questionnaire was used to collection the information about basic demographic characteristics, chronic disease history and physical activity prevalence of the study subjects. The body height and weight were measured by standard methods. Fasting venous blood samples were collected for the detection of fasting blood glucose, blood lipids and other biochemical indicators. Software SPSS 20.0 was used fort complex sampling sample weighting and data analysis. Results: The proportions of the study subjects who did regular exercise and never had leisure-time activity in 18-64 year-old labor force population in Beijing were 23.74% (95%CI: 21.51%-26.12%) and 64.34% (95%CI: 61.66%-66.94%) respectively; The average daily physical activity time was 23.44 min (95%CI: 21.16-25.71), and the total daily sedentary behavior time was 6.96 hours (95%CI: 6.80-7.13). With the increase of age, the proportions of the study subjects who did regular exercise and never had leisure-time activity increased (P=0.006, P<0.001). With the increase of educational level, the proportion of those who did regular exercise increased (P<0.001), and the proportion of those who never had leisure-time activity decreased (P<0.001); the higher the level of physical activity, the higher the regular exercise proportion (P<0.001); the proportions of employed people who did regular exercise was lower than those in unemployed people (P<0.001); the proportions of women never had leisure-time activity was higher than those in men (P=0.024). The results of multivariate analysis showed that older age and higher education level were positive factors for regular exercise (35- years old: OR=0.653, 95%CI: 0.530-0.804, P<0.001; 50-64 years old: OR=0.695, 95%CI: 0.560-0.864, P=0.001; high school/technical secondary school/technical School: OR=0.679, 95%CI: 0.593-0.777, P<0.001; college graduation: OR=0.478, 95%CI: 0.387-0.590, P<0.001; bachelor degree and above: OR=0.435, 95%CI: 0.347-0.546, P<0.001), while employment (OR=1.631, 95%CI: 1.330-2.000, P<0.001) and married/cohabitation (OR=1.340, 95%CI: 1.093-1.644, P=0.038) were negative factors for regular exercise. Conclusions: The rate of regular exercise in 18-64 year-old labor force population in Beijing needs to be improved. Older age and high educational level were positive factors for regular exercise, while being married and employed were negative factors for it.
Adolescent ; Adult ; Beijing ; Employment ; Exercise ; Female ; Humans ; Leisure Activities ; Male ; Middle Aged ; Sedentary Behavior ; Young Adult