Objective To establish a method of LC-MS/MS for determining cordycepin(Cor)and 3′-deoxyinosine(3′-Deo)concentration in rat plasma,and to study their pharmacokinetics in rats.Methods Protein was precipitated with methanol using 2-chloadenosine(2-Chl)as an internal standard.The chromatography was performed on Kinetex C18(3 mm×100 mm,2.6 μm,Phenomenex,USA)with gradient elution in aqueous(5 mmol·L-1 ammonium acetate)-methanol solution as mobile phase.ESI ion source was used for mass spectrometry,and positive ion multiple reaction monitoring(MRM)was used for scanning detection.The pharmacokinetics of Cor and 3′-Deo after oral administration of Cor(10 mg·kg-1)were studied in rats.Results Cor at 0.5-100 ng·mL-1 and 3′-Deo at 1-200 ng·mL-1 had good linearity,and the lower limits of quantification were 0.5 and 1 ng·mL-1,respectively.After oral administration of Cor in rats,the plasma concentration of Cor was low,which was mainly converted into the metabolite 3′-Deo.The Cmax of Cor and 3′-Deo were(5.4±3.4)and(142.0±50.0)ng·mL-1,and AUC0-360min min were(658.4±459.3)and(18 034.9±4 981.1)ng·min·mL-1,respectively.Conclusion The method is simple,sensi-tive,and accurate,which is suitable for determining Cor and 3′-Deo concentration in plasma and the pharmacokinetic study.

Objective To explore the factors influencing enteral nutrition intolerance in critically ill patients and to develop a risk prediction model to provide medical staff with a tool for early identification of patient intolerance.Methods Domestic and international databases such as CNKI,PubMed and Web of Science were searched and supplemented by searching references and grey literature.The search period was from inception to November 2022.Data were independently screened and extracted by 2 systematically trained researchers,and the quality of the literature was evaluated.Meta-analysis was performed using Review Manager 5.4 software.The OR value of the comprehensive effect of the factors was taken as the coefficient of each factor in the formula,and the natural logarithm of the ratio of intolerance incidence and non-incidence was the constant term of the formula.From December 2022 to June 2023,360 patients who met the inclusion and exclusion criteria in the ICU of a tertiary hospital were collected as a model verification group by convenient sampling method,and the collected clinical data were substituted into the formula to evaluate the discrimination and calibration of the model.Results A total of 13 articles were included.7 influencing factors with more than 3 times of reports and statistically significant results were obtained.For age[OR=0.97,95％CI(0.94,0.99),P=0.010],Acute Physiology and Chronic Health Evaluation scoreⅡ[OR=1.17,95％CI(1.01,1.36),P=0.040],comorbidity with diabetes[OR=1.21,95％CI(1.05,1.40),P=0.008],comor-bidity with neurological diseases[OR=0.85,95％CI(0.74,0.98),P=0.020],mechanical ventilation[OR=3.21,95％CI(1.82,5.66),P＜0.001],using sedative analgesics[OR=2.27,95％CI(1.66,3.10),P＜0.001],using gastric motility drugs[OR=0.23,95％CI(0.15,0.36),P＜0.001].The incidence of enteral nutrition intolerance was 35.00％.The risk prediction model for enteral nutrition intolerance in critically ill patients was logit(P)=-0.619-0.031×age+0.157 × APACHE Ⅱ+0.191 ×comorbidity with diabetes-0.163 ×comorbidity with neurosurgery+0.820 ×using sedatives and analgesics+1.166×mechanical ventilation-1.470×using gastric dynamic drugs.The area under the receiver operating characteristic curve of the model was 0.864.The maximum Youden index was 0.589.The sensitivity was 0.922.The specificity was 0.667.The corresponding clinical diagnostic threshold was 0.536.Hosmer-Leme-show test x2=13.410,P=0.098.Brier score was 0.195.Conclusion The risk prediction model of enteral nutrition intolerance in critically ill patients based on large sample evidence-based medicine is universal,scientific and practical.It provides a tool for medical staff to identify patients with enteral nutrition feeding intolerance in ICU.

Objective:To construct a perioperative activity program for patients with minimally invasive esophageal cancer under enhanced recovery after surgery (ERAS) concept through literature review and Delphi method.Methods:This was a single-center and interview study. Databases and websites including PubMed, Web of Science, the Cochrane Library, China National Knowledge Infrastructure, Wanfang Database, Medive and other databases and websites were searched systematically for articles related to esophageal cancer and upper digestive tract cancer patients′ activities from January 1, 2002 to January 31, 2022. In this single-center and interview study, literature review and group discussion were conducted to form a preliminary activity program, and then Delphi method was used to further demonstrate the practicability and feasibility of the activity program.Results:The overall program and specific intervention methods of perioperative activity program for minimally invasive esophageal cancer patients under ERAS concept were constructed, including 3 first-level indicators, 9 second-level indicators and 35 third-level indicators.Conclusions:Perioperative activity program of patients with minimally invasive esophageal cancer is scientific and reasonable, which can provide reference for medical staff and patients.

OBJECTIVE To explore the improvement effect and potential mechanism of Baicao fuyanqing suppository on bacterial vaginitis （BV） in rats. METHODS The female SD rats were randomly divided into normal group， model group， metronidazole group （positive control， 0.03 g/kg）， Baicao fuyanqing suppository low-dose， medium-dose and high-dose groups （0.18， 0.36， 0.72 g/kg）， with 8 rats in each group. Except for the normal group， the rats in other groups were injected subcutaneously with 0.2 g of Estradiol benzoate injection+20 μL of Escherichia coli suspension （2×108～3×108 cfu/mL） through the vaginal opening to establish the BV rat model. Administration groups were given relevant medicine vaginally， while the normal group and the model group were given normal saline， once a day， for 6 consecutive days. Twenty-four hours after the last administration， the vaginal appearance score and vaginal pH were measured for each group of rats. The levels of cytokines [interleukin-1β （IL-1β）， IL-2， IL-13， immunoglobulin A （IgA）] in vaginal lavage fluid were determined. The morphology of the uterus and accessories， and pathological changes in the vaginal tissue were observed. The expressions of Toll-like receptor 2 （TLR2）， TLR4 and nuclear factor-κB （NF-κB） in vaginal tissues were determined. RESULTS Compared with the normal group， the uterus edema， the irregular shape of uterus and accessories， the vaginal mucosa hyperplasia， and the massive desquamation of epithelial cells were observed in the model group， complicating with massive infiltration of inflammatory cells； vaginal opening redness and swelling score and secretion score， vaginal pH， the levels of proinflammatory cytokine IL-1β and IL-2， the protein expressions of TLR2， TLR4 and NF- κB were all increased or up-regulated， while the levels of IgA and anti-inflammatory cytokine IL-13 decreased significantly （P＜0.05 or P＜0.01）. Compared with the model group， varying cn degrees of improvement in uterine and accessories， and vaginal tissue lesions in rats were observed in administration groups， and most of the quantitative indicators mentioned above showed significant improvement （P＜0.05 or P＜0.01）. CONCLUSIONS Baicao fuyanqing suppository has a certain improvement effect on inflammatory symptoms in BV rats， and its mechanism may be related to the inhibition of the TLR/NF-κB signaling pathway.

Abstract: Objective To investigate the prevalence and influencing factors of depressive symptoms among primary and middle school students in Xihu District, Hangzhou City, so as to provide insights into depression control among primary and middle school students. Methods: Students were sampled from two primary schools, two junior high schools, two ordinary high schools and one vocational high school in Xihu District using a stratified cluster random sampling method from September to November, 2021. Participants' demographics, diet, exercise, sleep and campus bullying were collected using questionnaire surveys. The depressive symptoms were evaluated using the Chinese version of the Center for Epidemiologic Studies-Depression Scale (CES-D), and factors affecting the depressive symptoms were identified among primary and middle school students using a multivariable logistic regression model. Results: A total of 1 518 students were investigated, including 518 primary school students (34.12%), 363 junior high school students (23.91%), 463 ordinary high school students (30.50%) and 174 vocational high school students (11.46%). Of all participants, there were 805 males (53.03%). The prevalence of depressive symptoms was 38.87% among the participants, and was 36.10%, 42.42%, 42.33% and 30.46% among primary school, junior high school, ordinary high school and vocational high school students, respectively. The multivariable logistic regression analysis showed that gender (OR=1.601, 95%CI: 1.289-1.989), type of school (junior high school, OR=1.299, 95%CI: 0.974-1.734; ordinary high school, OR=1.351, 95%CI: 1.025-1.780; vocational high school, OR=0.862, 95%CI: 0.585-1.272), frequency of fresh vegetable intake (less than once daily, OR=0.454, 95%CI: 0.304-0.679; once and more daily, OR=0.359, 95%CI: 0.235-0.548), duration spent on the internet (0.5 to 1 h daily, OR=1.447, 95%CI: 1.044-2.006; 1 h and longer daily, OR=1.456, 95%CI: 1.072-1.978), sufficient sleep (OR=0.525, 95%CI: 0.371-0.744), suffering from campus bullying (OR=1.992, 95%CI: 1.491-2.661) were associated with the development of depressive symptoms. Conclusions The prevalence of depressive symptoms is high among primary and middle school students in Xihu District. Gender, type of school, dietary behaviors, sleep duration and campus bullying are factors affecting the development of depressive symptoms.

Objective:To develop a simple, practical and repeatable ultrasound method to locate the muscle at the trigger point of female myofascial pelvic pain(MPP), which can provide imaging reference for clinical precision treatment.Methods:A total of 113 patients with suspected MPP who came to the Women′s Hospital School of Medicine Zhejiang University from September 1, 2021 to April 20, 2023 were prospectively selected. The gynecologist performed internal examination with index finger on some pelvic floor muscles (puborectalis, pubococcygeus, iliococcygeus, coccygeus) and pelvic wall muscles (piriformis and obturator internus) respectively, searched for the muscles where the pain trigger point was located, and scored the pain by referring to visual analogue scale (VAS) and numerical rating scale (NRS), and then referred the patients to the ultrasound department. The ultrasound doctor used transvaginal ultrasound to display the above muscle groups in real time for observation and appropriate pressure. The muscle where the painful trigger point was located was found through tenderness and the pain score was performed. The two scores were compared for consistency and difference analysis.Results:The trigger point was clear and of good reproducibility. For the location and score of pain trigger points located in bilateral puborectalis, pubococcygeus and coccygeus, there was a strong consistency between the tenderness guided by vaginal ultrasound probe and clinical palpation (the consistency rate was ≥70%), and there was no significant difference in the pain scores of the trigger points located in the puborectalis muscle and coccygeal muscle between the two methods ( P>0.05), and there was statistically significant difference in the pain scores of the trigger points located in the other pelvic floor and pelvic wall muscles (all P<0.05). At the same time, ultrasonic examination made up for the deficiency of clinical palpation in the evaluation of piriformis muscle. Conclusions:The present method for finding the trigger point of MPP guided by the ultrasound probe is a new non-invasive, safe, simple and practical imaging method, which can provide a new imaging reference for the clinical diagnosis of MPP and the formulation of treatment strategies.

Sympathetic cues via the adrenergic signaling critically regulate bone homeostasis and contribute to neurostress-induced bone loss, but the mechanisms and therapeutics remain incompletely elucidated. Here, we reveal an osteoclastogenesis-centered functionally important osteopenic pathogenesis under sympatho-adrenergic activation with characterized microRNA response and efficient therapeutics. We discovered that osteoclastic miR-21 was tightly regulated by sympatho-adrenergic cues downstream the β2-adrenergic receptor (β₂AR) signaling, critically modulated osteoclastogenesis in vivo by inhibiting programmed cell death 4 (Pdcd4), and mediated detrimental effects of both isoproterenol (ISO) and chronic variable stress (CVS) on bone. Intriguingly, without affecting osteoblastic bone formation, bone protection against ISO and CVS was sufficiently achieved by a (D-Asp₈)-lipid nanoparticle-mediated targeted inhibition of osteoclastic miR-21 or by clinically relevant drugs to suppress osteoclastogenesis. Collectively, these results unravel a previously underdetermined molecular and functional paradigm that osteoclastogenesis crucially contributes to sympatho-adrenergic regulation of bone and establish multiple targeted therapeutic strategies to counteract osteopenias under stresses.
Adrenergic Agents/pharmacology* ; Apoptosis Regulatory Proteins/pharmacology* ; Bone Diseases, Metabolic/metabolism* ; Humans ; Liposomes ; MicroRNAs/genetics* ; Nanoparticles ; Osteoclasts ; Osteogenesis/physiology* ; RNA-Binding Proteins/pharmacology*

OBJECTIVE To analyze quality maker （Q-marker） of Ka nggongyan soft capsule （KSC）. METHODS The fingerprints of 20 batches of KSC were established by ultra high performance liquid chromatography （UPLC）method. Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition）were used to evaluate the similarity and confirm common peaks. The contents of norisoboldine ，leonurine hydrochloride ，forsythoside B ，acteoside，poliumoside and isoacteoside were determined by the same UPLC method. Targets and pathways related to KSC in the treatment of cervicitis were screened and analyzed by network pharmacology and molecular docking method to construct a “component-target-pathway”network，and analyze its potential Q-marker. RESULTS Twelve common peaks were identified in the fingerprints of 20 batches of KSC ，and the similarity was greater than 0.99. Six common peaks were identified ，including norisoboldine ，leonurine hydrochloride ，forsythoside B，acteoside，poliumoside and isoacteoside. The contents of the above 6 components were 1.336-1.774，0.093-0.143，4.970-5.888， 0.505-0.623，5.206-6.226 and 0.785-0.895 mg/g，respectively. By network pharmacology analysis ，14 key targets and 94 pathways were obtained ，and their binding energies to the core targets （protein kinase B 1，tumor necrosis factor ）were all less than －6.4 kJ/cal. CONCLUSIONS Six components such as norisoboldine and leonurine hydrochloride are potential Q-marker of KSC.

Objective:To investigate the correlation between subclinical hypothyroidism (SCH) and clinical outcomes of patients with acute ischemic stroke.Methods:From July 2014 to October 2017, patients with acute ischemic stroke admitted to Jiangsu Shengze Hospital Affiliated to Nanjing Medical University were enrolled retrospectively. Their demographic and baseline clinical and laboratory data were collected. The modified Rankin Scale was used to evaluate the clinical outcome at 3 months after the onset of symptoms. Multivariate logistic regression analysis was used to determine the independent correlation between SCH and clinical outcome of patients with acute ischemic stroke. Results:A total of 200 patients with acute ischemic stroke were enrolled, including 107 males (53.5%) and 93 females (46.5%). Their age was 69.67±11.38 years. There were 45 patients (22.5%) with SCH, 160 (80.0%) with good outcomes, and 40 (20.0%) had poor outcomes. Univariate analysis showed that there were significant differences in the baseline National Institutes of Health Stroke Scale (NIHSS) score, stroke etiology classification and the proportion of patients with SCH between the poor outcome group and the good outcome group (all P<0.05). Multivariate logistic regression analysis showed that high NIHSS score (odds ratio 2.884, 95% confidence interval 2.005-4.147; P=0.001) and SCH (odds ratio 19.527, 95% confidence interval 2.334-163.386; P=0.006) were the independent risk factors for poor outcomes. Conclusion:High NIHSS score and SCH were associated with the poor outcomes at 3 months after the onset of acute ischemic stroke.

In the present experimental study, cecal ligation and puncture significantly increased the myocardial injury assessed in terms of excess release of creative kinase-MB (CK-MB), cardiac troponin I (cTnI), interleukin (IL)-6 and decrease of IL-10 in the blood following 12 h of laparotomy procedure as compared to normal control. Also, a significant increase in protein expression levels of high-mobility group box 1 (HMGB1) and decreased phosphorylation of glycogen synthase kinase-3β (GSK-3β) was observed in the myocardial tissue as compared to normal control. A single independent administration of telmisartan (2 and 4 mg/kg) and AR-A014418 (1 and 2 mg/kg) substantially reduced sepsis-induced myocardial injury in terms of decrease levels of CK-MB, cTnI and IL-6, HMGB1, GSK-3β and increase in IL-10 and p-GSK-3β in the blood in sepsis- subjected rats. The effects of telmisartan at dose 4 mg/kg and AR-A014418 at a dose of 2 mg/kg were significantly higher than the telmisartan at a dose of 2 mg/kg and AR-A014418 1 mg/kg respectively. Further, no significant effects on different parameters were observed in the sham control group in comparison to normal. Therefore it is plausible to suggest that sepsis may increase the levels of angiotensin II to trigger GSK-3β-dependent signaling to activate the HMGB1/receptors for advanced glycation end products, which may promote inflammation and myocardial injury in sepsis-subjected rats.