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 study the influences of circular RNA ras homolog family member T1(CircRHOT1)on the prolifera-tion,apoptosis and immune escape of breast cancer cells by regulating the miR-187-3p/sex-determining region Y-box protein 4(SOX4)axis.Methods:Real-time quantitative PCR and Western blot were used to detect the expressions of CircRHOT1,miR-187-3p and SOX4 in human normal breast epithelial cells MCF-10A and breast cancer cells MCF-7,Hs-578T and MDA-MB-231 in vitro;MCF-7 cells cultured in vitro were randomly separated into control group,CircRHOT1 knockdown(transfected with CircRHOT1 siRNA plasmid)group,miR-187-3p mimics(transfected with miR-187-3p mimics)group,co-transfection negative control(transfected with empty plasmid and miR-187-3p inhibitor negative control)group,and CircRHOT1 knockdown+miR-187-3p inhibitor(transfected with CircRHOT1 siRNA plasmid and miR-187-3p inhibitor)group.After grouping and transfection,real-time quantitative PCR and Western blot were used to detect the expressions of CircRHOT1,miR-187-3p and SOX4 in each group;cell proliferation in each group was detected by EdU staining and plate colony formation assay;cell apoptosis in each group was detected by flow cytometry;im-munofluorescence staining was used to detect the ratio of apoptosis-related proteins Bax and Bcl-2(Bax/Bcl-2)expressions in each group.The cells of each group were co-cultured with human peripheral blood lymphocytes and transfected into groups,flow cytometry was used to detect the proportion of activated CD8+T cells in human peripheral blood lymphocytes in each group;the killing rate of human peripheral blood lymphocytes to MCF-7 cells in each group was detected by MTT assay.Dual-luciferase reporter assay was used to detect the targeted regulation of miR-187-3p and miR-187-3p on SOX4 by CircRHOT1 in MCF-7 cells.Results:Compared with human normal breast epithelial cells MCF-10A,human breast cancer MCF-7,Hs-578T and MDA-MB-231 cells had obviously higher protein and mRNA expression of CircRHOT1 and SOX4(P<0.05),and obviously lower expression of miR-187-3p(P<0.05).Com-pared with control group,the protein and mRNA expression of SOX4,EdU positive rate and colony formation rate of cells in Circ-RHOT1 knockdown group and miR-187-3p mimics group decreased(P<0.05),the expression of miR-187-3p,apoptosis rate,Bax/Bcl-2,the proportion of activated CD8+T cells in human peripheral blood lymphocytes and the killing rate on MCF-7 cells increased(P<0.05).Compared with CircRHOT1 knockdown group,there was no obvious difference in the expression of CircRHOT1 in the CircRHOT1 knockdown+miR-187-3p inhibitor group(P>0.05),the protein and mRNA expression of SOX4,EdU positive rate and colony formation rate of cells increased(P<0.05),the expression of miR-187-3p,apoptosis rate,Bax/Bcl-2,the proportion of activated CD8+T cells in human peripheral blood lymphocytes and the killing rate on MCF-7 cells decreased(P<0.05);there was no obvious difference in the indexes of cells in co-transfection negative control group(P>0.05).Conclusion:Knockdown of CircRHOT1 can reduce the expression of SOX4 by up-regulating miR-187-3p,thereby attenuating the proliferative activity of breast cancer cells and promoting their apoptosis.It can also inhibit the activation of CD8+T cells and enhance their cancer cell cytotoxicity,thereby weakening immune evasion of breast cancer cells.

Objective To assess the effect of ultrasonic thyroid artificial intelligence(AI)-assisted diagnostic system(AI-assisted diagnostic system)for diagnosing medullary thyroid carcinoma(MTC)compared with different physicians and taken papillary thyroid carcinoma(PTC)as the controls.Methods Totally 63 MTC,70 PTC and 62 benign thyroid nodules confirmed by pathology were enrolled.AI-assisted diagnostic system was utilized to analyze thyroid nodules and identify the likelihood of malignancy,and the probability value threshold was set at ≥0.40.All thyroid nodules were retrospectively reviewed and categorized by 3 physicians(1 senior physician,1 attending physician and 1 junior physician)according to Chinese thyroid imaging reporting and data system(C-TIRADS).The efficacy of AI-assisted diagnostic system and physicians for diagnosing MTC and PTC were evaluated.Results AI-assisted diagnostic system showed lower sensitivity,specificity,positive predictive value,negative predictive value,accuracy,and area under the curve(AUC)for diagnosing MTC and PTC compared with physicians.Significant differences of AUC were found between senior physician and AI-assisted diagnostic system,as well as between attending physician and AI-assisted diagnostic system for diagnosing MTC and PTC(all P＜0.01),while no significant difference of AUC was between junior physicians and AI-assisted diagnostic system(both P＞0.05).The sensitivity,specificity,positive predictive value,negative predictive value,accuracy and AUC for AI-assisted diagnostic system for diagnosing MTC were all lower than those for diagnosing PTC,but the AUC was not significantly different(P＞0.05).Conclusion Ultrasonic thyroid AI-assisted diagnostic system had relatively high value for diagnosing MTC.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

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.

Periodontitis is an inflammatory disease that is characterized by progressive destruction of the periodontium and causes tooth loss in adults.Periodontitis is known to be associated with dysbiosis of the oral microflora,which is often linked to various diseases.However,the complexity of plaque microbial communities of periodontitis,antibiotic resistance,and enhanced virulence make this disease difficult to treat.In this study,using metagenomic shotgun sequencing,we inves-tigated the etiology,antibiotic resistance genes(ARGs),and virulence genes(VirGs)of periodontitis.We revealed a significant shift in the composition of oral microbiota as well as several functional pathways that were represented significantly more abundantly in periodontitis patients than in con-trols.In addition,we observed several positively selected ARGs and VirGs with the Ka/Ks ratio＞1 by analyzing our data and a previous periodontitis dataset,indicating that ARGs and VirGs in oral microbiota may be subjected to positive selection.Moreover,5 of 12 positively selected ARGs and VirGs in periodontitis patients were found in the genomes of respiratory tract pathogens.Of note,91.8％of the background VirGs with at least one non-synonymous single-nucleotide polymorphism for natural selection were also from respiratory tract pathogens.These observations suggest a potential association between periodontitis and respiratory infection at the gene level.Our study enriches the knowledge of pathogens and functional pathways as well as the positive selection of antibiotic resistance and pathogen virulence in periodontitis patients,and provides evidence at the gene level for an association between periodontitis and respiratory infection.

Objective:To study the efficacy and safety of CT guided percutaneous transhepatic microwave ablation (PTPMWA) for primary liver cancer (PLC) in liver segment 9.Methods:A retrospective study was conducted on PLC patients between October 2013 and March 2019 at Dongguan People’s Hospital, Southern Medical University. Of 41 patients who entered into the study, there were 36 males and 5 females, with an average age of 59.1 years. These patients were diagnosed to have PLC in segment 9. The surgical related data and follow-up results were collected and analyzed.Results:All patients enrolled in the study completed the treatment procedure. CT scan was performed immediately after ablation which showed that the tumor areas to be completely covered by ablation. The duration of operation ranged from 45 to 260 (mean 91) min. The amount of bleeding during treatment was 1.0 to 5.0 (mean 1.4) ml. The complete response rate was 97.6% (40 patients) and the partial response rate was 2.4% (1 patient). The cumulative survival rates at 1, 2, 3, 4 and 5 years were 95.1%, 85.4%, 75.3%, 45.2% and 45.2%, respectively. Only 4 patients (9.8%) developed recurrence after treatment. The timings of recurrence were 1, 6, 13 and 67 months after treatment, respectively. The recurrent lesions were ablated again and complete response was obtained in all patients. There were no serious problems related to complications from ablation. The rate of postoperative complication was 7.3% (3 patients).Conclusion:PTPMWA is a novel treatment for patients with PLC in liver segment 9, the advantages of this treatment include good safety, high efficacy, low complications and local recurrence. The treatment is worthy of further future studies.

Objective To analyze the risk factors of gastrointestinal dysfunction in critically ill patients and provide reference for the prevention and treatment of gastrointestinal dysfunction. Methods A retrospective study was conducted, and the data of patients admitted to intensive care unit (ICU) of Jinghai District Hospital from September 2018 to March 2019 were collected. The data including sex, age, sequential organ failure score (SOFA), acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ), diagnosis in ICU, application of special drugs, hemoglobin (Hb), blood glucose, albumin (Alb) levels and presence or absence of bacteremia were collected. The patients were divided into gastrointestinal dysfunction group and non-gastrointestinal dysfunction group according to whether gastrointestinal dysfunction occurred or not. The general data, related final outcome and prognosis were compared between the two groups. Logistic regression analysis was used to analyze the risk factors affecting gastrointestinal dysfunction in critical ill patients, and the possible existing complications were recorded. The receiver operating characteristic curve (ROC) was drawn to evaluate the predictive values of risk factors. Results One hundred and thirty-eight patients were enrolled in this study, and 86 patients had gastrointestinal dysfunction. The SOFA score and proportions of using catecholamine and bacteremia in the gastrointestinal dysfunction group were significantly higher than those in the non-gastrointestinal dysfunction group [SOFA score: 7.2±3.8 vs. 5.8±3.6, the proportion of using catecholamine: 57.0% (49/86) vs. 38.5% (20/52), the proportion of bacteremia: 32.6%(28/86) vs.17.3%(9/52), all P < 0.05], Alb level was significantly lower than that in the non-gastrointestinal dysfunction group (g/L: 24.15±5.75 vs. 26.55±5.68, P < 0.05). Logistic regression analysis showed that the use of catecholamine, Alb level, bacteremia and SOFA score in ICU were the risk factors for occurrence of gastrointestinal dysfunction in ICU patients [odd ratios (OR) were 1.128, 0.547, 1.645, 1.958, 95% confidence intervals (95% CI) were 1.052-1.219, 0.384-0.765, 1.143-2.597, 1.925-1.993, P values were 0.011, 0.017, 0.021, 0.016, respectively]. Compared with the non-gastrointestinal dysfunction group, the incidence of bedsore, the proportion of energy intake unable to reach the target, the length of stay in ICU and the mortality in gastrointestinal dysfunction group were significantly increased [the incidence of bedsore: 53.5% (46/86) vs. 30.8% (16/52), the proportion of intake unable to reach the target: 27.9% (24/86) vs. 5.8% (3/52), the length of stay in ICU (days): 22.5±17.8 vs. 16.0±11.5, mortality rate: 51.2% (44/86) vs. 34.6% (18/52), all P < 0.05]. ROC curve analysis showed that the use of catecholamine, bacteremia present or not, Alb level and SOFA score showed certain extents of predictive values for the occurrence of gastrointestinal dysfunction in critically ill patients the area under ROC curve (AUC) were 0.794, 0.712, 0.705 and 0.882, respectively, 95% confidence interval (95% CI) were 0.708-0.880, 0.609-0.816, 0.579-0.830, 0.801-0.962, sensitivity were 58.8%, 42.5%, 76.3%, 75.0%, specificity were 100%, 60%, 100%, 85%, all P < 0.05. Conclusions The use of catecholamine, Alb level, bacteremia and high SOFA score are the risk factors of gastrointestinal dysfunction in critically ill patients. Prevention of gastrointestinal motility disorder can improve the treatment success rate of critical patients.

OBJECTIVETo compare the effects of moxibustion combined with highly active antiretroviral therapy (HAART) and simple HAART for human immunodeficiency virus (HIV) infected patients.

METHODSA total of 100 patients with HIV receiving HAART were randomized into an observation group and a control group, 50 cases in each one. In the observation group, moxibustion was used at Zusanli (ST 36), Guanyuan (CV 4) and Sanyinjiao (SP 6), etc. combined with HAART of zidovudine, lamivudine, nevirapine and efavirenzone, etc. Simple HAART was used in the control group. The patients were observed for 18 months. The indexes were CD, CD/CD, interleukin 2 (IL-2), interleukin 7 (IL-7), the incidence of side effects and the score of quality of life.

RESULTSAfter treatment, CD, CD/CD, serum IL-2 and the scores of quality of life (physiological, psychological, social relation fields and comprehensive score) increased and serum IL-7 decreased compared with those before treatment in the two groups (<0.01,<0.05), with better results except CDin the observation group (<0.01,<0.05). The incidences of gastrointestinal side effects and total side effects in the observation group were lower than those in the control group (14% (7/50) vs 32% (16/50), 58% (29/50) vs 80% (40/50), both<0.05).

CONCLUSIONMoxibustion combined with HAART for HIV infected patients could reduce the incidence of side effects, improve medication compliance, CD/CD, IL-2, IL-7 and the quality of life.

Objective To investigate the pattern of nodal recurrence after curative resection in adenocarcinoma of the gastroesophageal junction ( AGE ) , and to provide a basis for delineation of the radiation range in the high-risk lymphatic drainage area.Methods A retrospective analysis was performed in 78 patients with locally advanced AGE who were newly treated in our hospital from January 2009 to December 2013 and had complete clinical data.All patients received curative resection and were pathologically diagnosed with stage T3/T4 or N (+) AGE.Those patients were also diagnosed with SiewertⅡor Ⅲ AGE by endoscopy, upper gastroenterography, macroscopic examination during operation, and pathological specimens.None of the patients received preoperative or postoperative radiotherapy.All patients were diagnosed by imaging with postoperative nodal recurrence.The computed tomography images of those
patients were accessible and had all the recurrence sites clearly and fully displayed.Results The median time to recurrence was 10 months ( 1-48 months) , and 90%of the recurrence occurred within 2 years after surgery.The lymph nodes with the highest risk of recurrence were No.16b1( 39%) , No.16a2( 37%) , No.9 (30%), and No.11p (26%), respectively.There was no significant difference in the recurrence rate within each lymphatic drainage area between patients with SiewertⅡandⅢAGE ( P=0.090-1.000) .The lymph nodes with the most frequent recurrence were No.16b1, No.16a2, No.9, No.16b2, No.11p, and No.7 in patients with stage N3 AGE and No.11p, No.16b1, No.16a2, No.9, No.8, and No.7 in patients with stage non-N3 AGE.Patients with stage N3 AGE had a significantly higher recurrence rate in the para-aortic regions (No.16a2-b2) than those with stage non-N3 AGE (67%vs.33%, P=0.004, OR=4.00, 95% CI=1.54-10.37) .Conclusions The lymph nodes with the highest risk of recurrence are located in the celiac artery, proximal splenic artery, and retroperitoneal areas ( No.16a2 and No.16b1) in patients with SiewertⅡorⅢlocally advanced AEG.Moreover, patients with stage N3 AGE have a higher risk of retroperitoneal recurrence.The above areas should be involved in target volume delineation for postoperative radiotherapy.