Exosome is nanoscale vesicles with bilayer membrane structure, is secreted by multiple living cells in vivo, capable of carrying biological materials such as nucleic acids, proteins and lipids etc.Exosome can protect the information material (such as miRNA), which is easy to be inactivated or degraded in the circulatory system, so that they can be safely transferred to the specific target cells to participate in signal regulation, and play an important role in the information transmission between cells.Exosome is widely found in human blood, urine, pleural effusion and other body fluid samples, and can provide rich, stable, sensitive and specific biological information.As a kind of liquid biopsy specimen, exosome has a very high clinical application value in the future.

Objective The effect of serum exosome isolation method on obtained exosomal RNA is not yet clear .The aim of this study was to provide evidences to selected exosome isolation methods .Methods This was a method comparison study to assess three methods .Vein blood samples were collected from 3 healthy donors from Nanfang hospital , 4 ℃ 12 h was taken to make blood natural coagulated and the supernatant was taken as the serum . Exosomes extracted by Ultracentrifugation , ExoQuick and Total Exosome Isolation Reagent ( TEI ) kits from serum were assessed by transmission electron microscopy , nanoparticle tracking analysis and protein analysis to identify morphology , protein expression , concentration and size distribution of particles .ExoRNA extracted by Trizol-LS, SeraMir, Total Exosome RNA Isolation ( TER) and exoRNeasy were assessed by Bioanalyzer 2100 and qPCR to ascertain RNA distribution and miRNA expression.Results For exosomes isolation , two commercial kits ( ExoQuick and TEI ) showed higher exosomes recovery and protein concentration than ultracentrifugation , but exosome isolated by ultracentrifugation expressed abundant protein makers mostly .For exo-RNA isolation, the distribution of RNA and expression of miRNAinfluenced by three methods , but there was no effect on the relative expression trend of miRNA.ExoRNeasy resulted in the highest yield and most narrow size distribution pattern of small RNA with higher level of miRNA expression .Results of TEI with TER kits showed no obvious bands of small RNA and moderate miRNA expression among methods .Ultra with Trizol-LS or ExoQuick with SeraMir showed low concentration measurable bands around 100 nt, with changeable miRNA profiling irregularly . Conclusions Extraction of exosomes using traditional ultracentrifugation method is applicable for proteomics research work .Extraction using ExoQuick and TEI kits is suitable for preparing exosomes from scarce samples such as serum.

Objective To observe the expression level and diagnosis performance of serum exosomal miR-21 in lung cancer. Method Sera of 94 lung cancer patients,29 pulmonary tuberculosis patients,and 26 healthy controls from Nan Fang Hospital were collected. Exosomes in serum were purified and characterized ,and miRNA was abstracted from exosomes. Levels of exosomal miR-21 in three groups were compared with relative quantitative RT-PCR. Diagnosis performance of exosomal miR-21 was evaluated by ROC curve. Results Purified serum exosome sample possesses typical exosome features. There were significant difference among exosomal miR-21 levels from lung cancer ,tuberculosis ,and health control group. Exosomal miR-21 obviously increased in lung cancer patients,and decreased in tuberculosis patients. Exosomal miR-21 were increased in early and late stage lung cancer patients. The diagnostic performance of exosomal miR-21 was evaluated by ROC curve. Its area under the curve was 0.702 and specificity was 0.923. Conclusions Serum exosomal miR-21 was increased in many kinds of lung can-cer patients. It possesses excellent diagnosis performance ,which may be developed as new biomarkers to assist lung diagnosis.

Objective To investigate the impact of vascular inflow occlusion of the liver on arterial lactate level and pH value. Methods In this study, 68 patients who underwent hepatectomy from January 2006 to December 2008 were retrospectively studied. The patients were assigned to one of the three study groups according the vascular inflow status: clamping of portal vein and hepatic artery in the hepatic pedicle (n = 20), hemihepatectomy under total hemihepatic vascular exclusion (THVE, n = 22), and non-vascular occlusion (n = 26). Postoperative arterial blood gas analysis including systemic arterial lactate concentration, and liver and renal function tests were performed. Results Systemic arterial blood lactate levels significantly elevated in the portal clamping and THVE groups (5.53 ±2. 31 mmol/L and 5.62 ±2.52mmol/L, respectively), compared to the non-occlusion group (3. 37 ± 1.56 mmol/L, P < 0. 05) ;significant increase in arterial HCO3- level was observed in the THVE group in comparison to the nonocclusion group (19. 68 ± 3. 82) mmol/L vs. (21.65 ± 2. 48) mmol/L, (P < 0. 05). There were no significant differences as to the changes of pH values, liver and renal function tests between the three groups. Conclusions Vascular inflow deprivation may result in significantly increased arterial lactate level. Thus, intense surveillance of blood lactate level with prompt treatment is necessary to prevent postoperative hyperlactatemia and metabolic acidosis.

Objective To summarize the clinical diagnosis and treatment of nodular regenerative hyperplasia of the liver. Methods Retrospective analysis was made on the clinical manifestations,imagings, laboratory tests, diagnosis, treatment and prognosis of 18 consecutive cases finally established as NRH during the past 26 years. Results 15 of the 18 cases showed portal hypertension, 4 cases showed mono or multiple occupations of the liver, 8 cases suffered from concurrent autoimmune diseases, 3 cases were suspected of blood diseases. Preoperatively, 13 cases were diagnosed as cirrhosis, 2 cases were diagnosed as liver cancer or focal nodular hyperplasia ( FNH). All cases were diagnosed by operative wedging biopsy. 3 cases received splenectomy, 4 cases received disconnection /Phemister surgery, 3 cases received liver occupation/liver lobe resection, 1 case received partial small bowel resection, and 1 case received spleen artery restrictive surgery. Postoperatively, symptoms of portal hypertension relieved obviously. Follow-up study showed most of the patients were stable and prognosis of the NRH was good.Conclusions NRH may relate to the disturbance of liver blood supply, and most common clinical manifestation is portal hypertension, and can combine with immune diseases, hematopathy also can present single or multiple liver occupations. Differential diagnoses include liver cirrhosis, FNH, idiopathic portal hypertension. Diagnosis of NRH relies on liver wedging biopsy. Surgery can relive concurrent portal hypertension.

Objective To investigate the effect and long-term prognosis of patients who underwent hepatectomy for hepatolithiasis. Methods We retrospectively analyzed the medical records of 98 patients with hepatolithiasis who were treated by hepatectomy in Peking Union Medical College Hospital.Results Male/femah:1/1.7;median age:55 years old.58 cases(59.2%)had been treated before;among them.50 by surgery.In 88 cages(89.8%)hepatolithiasis involved the left lobe only,in 2(2.0%) only the right lobe involved.and in 8(8.2%)both left and right lobe were involved.51(52.0%)had extrahepatic biliary stones,30(30.6%)had biliary duct strictures,28(28.6%)had a history of biliary ascariasis.and 5(5.1%)had a concurrent biliary tract malignancy.All received partial hepatectomy according to the stone location;for the 8 bilaterally involved patients,left hepatectomy and right lithotomy were performed.Postoperative complications occurredin 14 cases(14.3%),and there were2perioperative deaths(2.0%).Seventy-eight patients(79.6%)have beenfollowedupfor over1 yearwith no tumors;the results were excellent or good in 91.0%;the stone residue and recurrence rate were both 2.0%.Conclusions Hepatectomy not only eliminates calcuci,but also removes diseased biliary tracts,with advantages of low residue stone and recurrence rate.

Objective To investigate the expression change of cytokines in peripheral blood and intestinal mucosa in the patients with diarrhea post-infectious irritable bowel syndrome(PI-IBS) and its relation with clinical symptoms scores.Methods Thirty outpatients and inpatients with diarrhea PI-IBS(observation group) and contemporaneous 30 individuals undergoing physical examination(control group) in the Hainan Provincial People's Hospital from January to December 2013 were selected.The peripheral blood mononuclear cells(PBMC) were separated and cultured.Then the levels of IFN-y and IL-10 in peripheral blood and cell culture supernatant fluid were detected by ELISA.The colonic mucosal tissue was taken by coloscopy.Then colonic mucosal IFN-γ and IL-10 protein expression was detected by immunohistochemistry staining.Furthermore,the correlationship between the level change of IFN-γ and IL-10 with clinical symptom score was analyzed by using the Spearman correlation method.Results Peripheral blod IL-10 and IFN-γ levels had no statistical difference between the two groups(P＞0.05).Compared with the control group,in PBMC seperation and cuture,the IFN-γ level in the observation group was increased and IL-10 level was decreased,the difference was statistically signifieant(P＜0.01).The intestinal main symptom score in the observation group had the positive correlation with IFN-γ expression level of PBMC culture supernatant fluid and colonic mucosal IFN-γ expression level(r=0.45,0.94,P＜0.01),and had the negative correlation with IL-10 expression level(r=-0.52,-0.79,P＜0.01).Conclusion The unbalance of IFN-γ and IL-10 level could be involved in the pathogenesis of diarrhea PI-IBS,which can serve as the observation indicators of disease activity.

Objective: To explore the safety and feasibility of associating diaphragm resection and liver-diaphragmatic metastasis lesions resection for patients with advanced ovarian cancer. Methods: Retrospectively analysis 83 cases(98 times) of advanced ovarian cancer with liver-diaphragmatic metastasis between January 2012 and December 2016 at Department of Liver Surgery, Peking Union Medical College Hospital.The patients were aged from 19 to 75 years.Surgical procedure included metastatic lesions resection(43 times) and stripping(55 times). Operation status, post-operative complications, pathology results and follow-up of the patients were analyzed. Results: Fifteen patients received twice surgical treatment and 68 patients received one time surgical treatment. Postoperative hemorrhage in chest and between liver and diaphragm was not occurred in all cases.Dyspnea and low oxygen saturation were occurred in two cases of stripping patients and 1 case of metastatic lesions resection patients.Results of CT examination indicated that there was medium to large amount of ascites in right chests.The symptoms were relieved after placing thoracic closed drainage.Other patients were recovered smoothly.All patients were diagnosed as ovarian cancer by pathological examination. Conclusion Associating diaphragm resection is safe and feasible for liver-diaphragmatic metastasis lesions from ovarian cancer.

Objective: To investigate the effects of epidural analgesia on puerpera and neonatal. Methods: A total of 150 puerpera were continuously enrolled from May 2017 to May 2018 in Zhoushan Maternal and Child Health Care Hospital, and were divided into analgesia group (102 cases) and non-analgesia group (48 cases) according to their prenatal wishes.The analgesia group was given epidural anesthesia for analgesic delivery, while the control group had no analgesia intervention.Differences in the expression of serum pain and stress hormones were compared between two groups at the time of prenatal, postpartum 1h and 24h.At the same time, differences in neonatal NACS scores were compared between two groups at the time of postpartum 1h, 8h and 24h. Results: No statistically significant differences were found between the two groups in prenatal serum pain and stress hormones, but at the time of postpartum 1h and 24h, serum pain media neuropeptide Y (NPY) (t=5.041, P=0.000; t=4.984, P=0.000), 5-hydroxytryptamine (5-HT) (t=3.083, P=0.002; t=4.203, P=0.000), prostate E2 (PGE2) (t=6.764, P=0.000; t=4.364, P=0.000) and stress hormone angiotensin Ⅱ (AngⅡ) (t=3.670, P=0.000; t=2.911, P=0.004), adrenocorticotropic hormone (ACTH)[(212.54±38.65) ng/mL vs.(231.43±34.82)ng/mL, t=3.510, P=0.001; (163.82±33.21) ng/mL vs.(175.21±32.48) ng/mL, t=2.105, P=0.045)]and cortisol (Cor)[(344.56±57.39) μg/L vs.(381.81±59.23)μg/L, t=3.670, P=0.000; (241.27±43.44) μg/L vs.(262.56±37.97) μg/L, t=2.911, P=0.004)] in the analgesia group were significantly lower than those in the non-analgesia group.There were no statistically significant differences in NACS score, complications and Apgar score between the two groups (all P>0.05). Conclusion The analgesic effect of full-course epidural delivery in analgesic delivery is remarkable, which can effectively reduce maternal stress reaction, and it is safe and has less adverse effect on neonates.

OBJECTIVE To explore the value of providing pharmaceutical service related to risdiplam in direct-to-patient （DTP） pharmacies. METHODS The follow-up data of spinal muscular atrophy （SMA） patients who purchased and used risdiplam from Shangyao Yunjiankang Yiyao Pharmacy （Shanghai） Co.， Ltd. from May 2021 to January 2023 were collected. The medication information， therapeutic efficacy and the occurrence of adverse events were retrospectively analyzed. RESULTS A total of 42 prescriptions were checked by pharmacists in the DTP pharmacies， and 7 prescriptions were found to be unreasonable （16.7%， 7/42）， which were corrected after the timely intervention. During the follow-up management， pharmacists replied to 4 patients （9.5%， 4/42） regarding medication consultation about medication requirements and adverse events. Two patients with type Ⅰ SMA experienced adverse events： one of them presented with fever and the other presented with skin dryness with darkening. Both of them were grade Ⅰ toxic reactions and generally did not require clinical treatment. Considering that the patient sustained low-grade fever for a long time， the pharmacist suggested symptomatic treatment under the guidance of the doctor. CONCLUSIONS Pharmacists in DTP pharmacies conducting follow-up management of risdiplam use for rare disease SMA patients can help promote rational， standardized medication for patients.