Objective To apply accelerated solvent extraction (ASE) technique to extract Achyranthes bidentata and to explore the application of this technique in quality control of Chinese materia medica. Methods Investigation of single factor was used to optimize the conditions that affected the efficiency for ASE from A. bidentata by RP-HPLC using ecdysterone as a quantitative marker. Results The optimized conditions for ASE of A. bidentata were obtained as follows: methanol as solvent, particle size between 0.3 and 0.45 mm, temperature at 100 ℃, pressure under 10.34 MPa, 6 min duration and once extraction. Conclusion ASE technique can be used to extract A. bidentata quickly and effectively.

ObjectiveThe expression and significance of interleukin(IL)-17 in a murine model of experimental systemic sclerosis(SSc) was studied and its correlation with transforming growth factor-beta 1 (TGF-β1) was explored.Methods Thirty female BALB/c mice were randomly divided into 3 groups,including a control group, bleomycin(BLM) injection for 4 weeks group(model 1 group) and a termination injection of BLM 4 weeks group(model 2 group).The pathological changes of skin and lung were detected.The mRNA expressions of IL-17A,RORγt,TGF-β1 mRNA were evaluated by real-time PCR.Enzyme linked immunosorbent assay was used to measure the levels of IL-17 and TGF-β1 in the serum and bronchoalveolar lavage fluid(BALF).Comparisons among groups were performed by variance analysis.ResultsSkin and lung of the model groups showed evident inflammatory cell infiltration and increased deposition of collagen fibers.The score of dermal inflammation and lung fibrosis was significantly higher in the model 1 and model 2 groups (2.5±0.8,3.0±1.8), (2.4±0.8,3.1±1.2) as compared to that of the control group (0.9±0.7,0.9±1.0),(F=12.19,8.367,25.11,4.641; all P＜0.05).The amount of hydroxyproline was markedly increased in the model groups than in the control group.Compared with those of the control group,the mRNA levels of IL-17A,RORγt,TGF- 31 in the skin and lung of the model 1 group were higher.The levels of IL-17 in serum and BALF of the model 1 group was significantly increased and the levels of TGF- β1 were increased in BALF and decreased in the serum (all P＜0.05).The mRNA levels of IL-17A in skin and lung had a positive correlation with the mRNA levels of TGF- β1,score of dermal inflammation and lung fibrosis.The levels of IL-17 in serum had a positive correlation with hydroxyproline of the skin and lung.ConclusionIL-17 may participate in systemic immune-mediated inflammation and changes of skin and lung in SSc and when combined with TGF-β1 togetter will cause damage to skin and lung in SSc.

Objective To study the expression and significance of Th17 and Tc17 cells in the peripheral blood,skin and lung in a murine model of bleomycin (BLM)-induced systemic sclerosis (SSc).Methods Thirty female BALB/c mouse were randomly divided into 3 groups,including a control group ( A group),a injected with BLM 4 week without pulmonary fibrosis(PF) group( B group) and with obviously PF group(C group).Pathological changes of skin and lung were detected.The proportion of CD4+,CD8+,CD4+IL-17+(Th17),CD8+IL-17+(Tc17) cells in the peripheral blood,skin and lung of mouse was determined by flow cytometry.The mRNA expressions of RORγt,IL-17A in skin and lung of mouse were evaluated by real-time PCR.Enzyme linked immunosorbent assay(ELISA) was used to measure the levels of IL-17 in serum.Results Dermal hydroxyproline(HYP) contents and the score of PF were significantly increased in C group [ (3.07±1.26) μg/mg,4.0±1.41 ]and B group [ (2.43±0.61) μμg/mg,1.50±0.76]as compared with A group [ (1.45±0.40) μg/mg,0.60±0.70 ],and pulmonary HYP contents was obviously increased in C group than in A and B groups,all P＜0.05.Compared with the A group,the percentage of CD4+ and Th17 cells in the peripheral blood,skin and lung of B and C groups,Tc17 cells of C group was significantly increased,and CD8+ cells was significantly decreased(all P＜0.05).The ratio of Th17/CD4+CD8+ in the peripheral blood,skin and lung of B and C groups [ ( 1.41 ±0.36)％,( 1.79±0.77)％ ],[ (2.58±1.07)％,(5.23±2.34)％ ]and [ (3.50±1.20)％,(4.02±1.32) ％ ]was significantly increased compared with A group (0.71±0.25)％,(1.15±0.59)％,(0.99±0.46)％.The ratio of Tc17/CD4+CD8+ in the lung of C groups( 1.62±0.53) ％ and in the skin of B and C groups [ (1.70±0.70) ％,( 1.63±0.63 ) ％ ]was significantly increased compared with A group [ ( 1.00±0.47 ) ％,( 1.1 1 ±0.34 ) ％ ],all P＜0.05.Compared with the A group,the mRNA levels of IL-17A,RORγt in skin of B and C groups,and in lung of C group were higher and the levels of IL-17 in serum was significantly increased,all P＜0.05.Th17 cells and the levels of IL-17 in blood were positive correlation with dermal and pulmonary inflammation,fibrosis and H YP contents,all P＜0.01.The frequency of Th17 and Tc17 cells in skin and lung respectively had a positive correlation with dermal and pulmonary inflammation,the score of fibrosis,and HYP contents of skin and lung,all P＜0.01.Conclusion Th17 and Tc17 cells were significantly increased in the peripheral blood,skin and lung of a murine model of SSc,and Th17 cells is dominated.They correlated with the inflammation and fibrosis of skin and lung,and may participate in the pathogenesis of SSc through secrete IL-17.

Objective To assess the mid-term and long-term efficacy of the permanent inferior vena cava filter in the treatment of deep vein thrombosis of lower limb and discuss the clinical significance of inferior vena cava filter.Methods Retrospectively analyze on the 86 cases with deep vein thrombosis of lower limb (41 males and 45 females,aged 50 to 94 years,mean age was 71.8 years) treated with implantation of permanent inferior vena cava filter in inferior vena cava from Janunary 2010 to October 2015.In these patients,there were 51 cases with embolism in the left leg,25 cases in the right leg,10 cases in both legs and 6 cases were accompanied with pulmonary embolism.The cases without contraindication underwent catheter directed thrombolysis and even percutaneous transluminal angioplasty or stents subsequently if necessary after inferior vena cava filter implantation.All the cases with no contraindication were treated with anticoagulant therapy.Results All the 86 patients were implanted inferior vena cava filter (B.Braun Vena Tech LP 76 and Cordis TrapEase 10)successfully.Sisty-five cases were underwent inferior wena cava filter implantation only,while 21 cases were treated with inferior vena cava filter implantation and catheter directed thrombolysis or even percutaneous transluminal angioplasty and stents.During the follow-up period(12 to 81 months,mean time was 51 months),27 patients died dueing to malignant tumor(17 cases) and other diseases (10 cases) rather than complications caused by inferior vena cava filter.Three patients had recurrence of deep vein thrombosis and 2 patients suffered from the thrombosis induced by stenosis of stents.Inferior vena cava filter appered tilted with angle less than 15 degrees in 6 cases.Three cases suffered from new thrombosis below the filter and 2 cases complained of the filter migration.No case was found with fracture of filter,perforation of the inferior vena cava,bleeding or pulmonary embolism(new onset or recurrent).Conclusions Application of permanent inferior vena cava filter may cause complications,though it is an effective approach to prevent pulmonary embolism in patients with deep vein thrombosis of lower limb.However,permanent inferior vena cava filter may be fit for patients with old age,incurable cancer or limited expected life.

Objective To explore the effect of low molecular weight heparin calcium on coagulation function and platelet(PLT)in patients before and after anterolateral thigh flap transplantation.Methods Fifty patients with skin and soft tissue defects of hands and feet treated in Wuxi Ninth Hospital and Jiujiang No.1 People's Hospital from January to December 2022 were selected as the study objects.They were divided into observation group(26 cases)and control group(24 cases)according to random number table method.Patients in observation group received low molecular weight heparin calcium 12h before surgery,and continued to receive routine treatment once a day after surgery.Patients in control group began to use low molecular weight heparin calcium routinely 4h after surgery,once a day.The coagulation function and PLT before and after surgery were compared between two groups.Results There were no significant differences in incidences of vascular crisis and skin flap necrosis between two groups(P>0.05).Five patients in observation group received intraoperative or postoperative blood transfusion,which was excluded in further analysis.On first day and third day after surgery,fibrinogen(FIB)and PLT in two groups were significantly higher than those before surgery,and thrombin time(TT)was significantly shorter than that before surgery(P<0.05).On third day after surgery,activated partial thromboplastin time(APTT)and prothrombin time(PT)in observation group were significantly longer than those before surgery(P<0.05).Before surgery,first day and third day after surgery,there were no significant differences in FIB,APTT,PT and TT between two groups(P>0.05).On third day after surgery,PLT of observation group was significantly higher than that of control group(P<0.05).Conclusion Both FIB and PLT were higher after surgery regardless of the use of low molecular weight heparin calcium before and after surgery,and FIB and PLT were more significantly higher in patients treated 12h before surgery.

Objective To discuss the effect of a moving folding multifunction operating platform in operation room so as to solve time-consuming and efforts in the putting lithotomy position of repeating placement and removal surgery bed legs plate and supply the operating platform in the surgery of the upper limb, inva-sive puncture and anesthesia operation.Methods A moving folding multifunction operating platform was self-designed. A number of 160 cases undergoing operation in lithotomy position in the urology department from March to June 2016 were randomly divided into the experimental group (80 cases) and the control group (80 cases). The routine nursing method were carried out in patients in the control group, while adopted operating platform were used in patients in the experimental group. Time-consuming pre and post operation were investigated. Application effects of moving folding multifunction operating platform in 114 cases including the surgery of the upper limb, invasive puncture and anesthesia operation were evaluated.Results During continues lithotomy position surgery, time consuming of changing position and position placement in the experimental group were (209.41±33.36) s and (622.33±62.16) s respectively, while those in the control group were (665.83±90.92) s and (1118.33±88.86) s. The differences were statistically significant between two groups (t=16.324, 15.844;P<0.05). In one lithotomy position surgery, time consuming of position placement pre and post operation were reduced 44.03% (saved 8.16 minutes/surgery) compared to that in the control group. It alleviated 60 kg body burden of nurses. The safety and performance evaluation in the surgery of the upper limb, invasive puncture and anesthe-sia operation were in high levels with scores of (26.13±1.53) and (27.42±1.56) respectively.Conclusions The platform, which is practical and easy to use, contributes to meet the needs of a variety of technical operation in the operation room and helps optimizing procedure in transformation, reducing work load of nurse so as to im-prove efficiency and quality of work.

Objective:To investigate the etiological spectrum, clinical features, and surgical treatment of patients with Cushing's syndrome (CS) who underwent adrenal surgery.Methods:From August 2002 to August 2022, the clinical data of 985 patients with Cushing's syndrome who underwent surgical treatment in the department of urology, Peking Union Medical College Hospital were retrospectively analyzed. There were 210 males and 775 females. The average age was 43.33±13.49 years old. The age of males was older than that of females (45.53±14.39 vs. 42.68±13.16 years, P=0.016). The principle of preoperative surgical method selection for patients in this group was described as follow. For adrenocorticotropin (ACTH) independent CS, adrenal tumor resection was considered for unilateral solitary lesions and unilateral adrenalectomy was considered for unilateral multiple lesions. For bilateral lesions, the larger tumor was removed first, and the contralateral operation was decided according to the follow-up results. Patients with suspicion of cortical cancer are subjected to R0 resection, and open surgery was performed if the tumor diameter is≥6 cm. The clinical characteristics of CS patients were summarized. The clinical symptom characters, etiology spectrum and the corresponding selection principles of surgical methods were analyzed. Results:Among the 985 cases, ACTH-independent CS accounted for 92.8% (914/985), and ACTH-dependent CS accounted for 7.2% (71/985). According to the postoperative pathological results, ACTH-independent CS was the most common, among which adrenal adenoma was the most common 75.94% (748/985), PBMAH 11.37% (112/985), Cushing's disease 4.26% (42/985), PPNAD 3.25% (32/985), EAS 2.94 (29/985), cortical carcinoma 2.23% (22/985). In terms of clinical manifestations, full moon face, bloody face, buffalo back, central obesity and weight gain were more common. The incidence of the above single symptoms was >30%. In terms of hormone secretion, the cortisol level of ACTH-dependent CS patients was significantly higher than that of ACTH-independent CS ( P<0.001). Cortisol [(16.61±6.78) μg/dl] and 24h-UFC [103.65 (59.83, 175.70) μg/24h] in patients with subclinical cortical adenoma were lower than those in other types of patients ( P<0.001). Among the patients with bilateral adenomas, 4 patients underwent simultaneous resection of bilateral adrenal tumors due to severe CS symptoms, and the remaining 74 patients underwent two-stage simple tumor resection, total adrenalectomy or subtotal adrenalectomy according to the size and number of tumors. Of the 22 patients with cortical carcinoma, 2 patients underwent R0 resection after neoadjuvant therapy with mitotane after being diagnosed by needle biopsy due to local infiltration of the tumor. In patients with EAS and Cushing's disease, 62.07% (18/29) and 23.81% (10/42) were treated with simultaneous bilateral target gland resection due to severe CS symptoms, respectively. In 112 cases of PBMAH and 32 cases of PPNAD, the initial treatment was unilateral adrenalectomy or subtotal adrenalectomy, and the follow-up was continued to decide whether to perform contralateral adrenalectomy or subtotal adrenalectomy. Conclusions:The etiological spectrum of CS patients admitted to the urology department is different from that of the overall CS, and is dominated by adrenal disease, and Cushing's disease is rare. Due to the different etiological spectrum, the patient's symptom spectrum is different, and difficulty in squatting and fracture are rare. The main treatment for unilateral adrenal disease is tumor resection or unilateral adrenalectomy. For ACTH-dependent CS in bilateral lesions, bilateral adrenalectomy is recommended, and hormone supplementation is recommended after surgery. For ACTH-independent CS, unilateral adrenalectomy is recommended first, and individualized treatment plans are formulated based on the results of follow-up.

Objective To explore the inflammatory effect of exosomes derived from alveolar epithelial cells stimulated by lipopolysaccharide (LPS) on the alveolar macrophages (NR8383). Methods The alveolar epithelial cells disposed with different treatments were co-cultured with alveolar macrophages by using a Transwell system separately. Alveolar epithelial cells (RLE-6TN) were randomly divided into 4 groups: normal group, LPS-stimulated group, exosome inhibitor group, and exosome inhibitor pretreatment + LPS stimulation group. NR8383 cultured alone was considered as a blank control. After the 12-h co-culture, the real-time PCR (qPCR) was performed to examine the mRNA relative expression of IL-6, TNF-α, and IL-1β in NR8383 cells. To further explore the role of exosomes derived from RLE-6TN on alveolar macrophages mediated inflammationary response, the experimental exosomes (exosomes derived from LPS-induced RLE-6TN) and control exosomes exosomes derived from normal RLE-6TN were extracted by gradient ultracentrifugation. Transmission electron microscopy and Western blotting analyses was performed to identify the exosomes, and qNano particle diameter analyzer was conducted to measure the particle diameter of exosomes. In vitro, NR8383 cells were divided into 3 groups which were cultured with exosomes derived from LPS-stimulated RLE-6TN at a concentration of 10 μg/mL (experimental group), exosomes derived from untreated RLE-6TN at the same concentration of 10 μg/mL (control group), and the PBS at the same volume with experimental group (PBS group), respectively for 12 h. After the treatment, the phagocytosis of NR8383 cells was observed by laser confocal microscope and the release of interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α in supernatants of NR8383 was detected by enzyme-linked immunosorbent assay ELISA Results (1)In the co-culture experiment, the mRNA relative expression of pro-inflammatory cytokine in the LPS group was significantly increased compared with the blank control group (P<0.01), however comparing the exosome inhibitor pretreatment+LPS group with the LPS group, the expression of pro-inflammatory cytokine was decreased (P<0.01). (2) The extracted exosomes were observed as circular or elliptical vesicles with a diameter of 40-100 nm under the transmission electron microscopy. Western blotting analyses showed that the extracted exosomes express the protein marker, such as CD63 and CD9; After incubation with NR8383 cells for 5 h, laser scanning confocal microscope showed that the exosomes labeled with red fluorescent were uptaken by NR8383 cells. (3)After the exosomes derived from the LPS-disposed RLE-6TN and the normal RLE-6TN cells were incubated with NR8383 cells respectively. The ELISA test showed that treated the alveolar macrophages with LPS induced alveolar epithelial secreted exosomes led to a robustly increased release of pro-inflammatory cytokine (P<0.01), but there was no significant difference between the control group and PBS group (P>0.05). Conclusions Exosomes derived from LPS-disposed alveolar epithelial cells activate the alveolar macrophage-mediated inflammatory response.

Objective To observe the amplification effect of mitochondrial DNA (mtNDA) on the inflammatory response of rat alveolar macrophage induced by lipopolysaccharide (LPS), and to preliminarily explore its mechanism. Methods mtDNA of Sprague-Dawley (SD) rat liver tissue was harvested, ultra-micro spectrophotometer and 1% agarose gel electrophoresis were used to detect the concentration and quality of mtDNA. The alveolar macrophages of SD rat were isolated and cultured, the macrophages in logarithmic growth phase were divided into phosphatic buffer solution (PBS) group, LPS group, mtDNA group and LPS+mtDNA group. The first three groups were added equal volumes of PBS, LPS 1 mg/L or mtDNA 10 mg/L to the alveolar macrophages medium for 6 hours and 12 hours, respectively; the alveolar macrophage medium of LPS+mtDNA group was stimulated with 1 mg/L LPS for 6 hours and then stimulated with 10 mg/L mtDNA for 6 hours. The levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the cell supernatant were detected by enzyme linked immunosorbent assay (ELISA);the expression of the key protein of Pyroptosis-Gasdermin D (GSDMD) was detected by Western Blot. Results ① The mtDNA A260/280 ratios were between 1.8-2.0, and agarose gel electrophoresis showed a single band, with a size of about 16 kb. ② After alveolar macrophages stimulated by LPS or mtDNA for 6 hours or 12 hours, respectively, the levels of IL-1β and TNF-α were higher than those in PBS group. When cells were treated with mtDNA for 6 hours after LPS stimulation 6 hours, the levels of IL-1β were higher than those in LPS 12 hours group (ng/L: 366.27±23.35 vs. 154.70±23.32, 1 < 0.01), but the levels of TNF-α had no significant difference compared with LPS 12 hours group (ng/L: 836.13±25.01 vs. 802.67±30.48, 1 > 0.05). ③ The protein expressions of GSDMD in LPS group, mtDNA group and LPS+mtDNA group were significantly higher than those in PBS group (GSDMD/β-actin: 1.77±0.05, 1.65±0.04,2.40±0.05, 1.00±0.02, all 1 < 0.01), and the protein expression of GSDMD in LPS+mtDNA group was higher than that in LPS group (1 < 0.01). Conclusion mtDNA amplifies LPS-induced alveolar macrophage inflammatory responses,which mechanism may be related to the increase in pyroptosis mediated by mtDNA.

Objective To evaluate the diagnostic efficacy of Kwak and ACR( 2017 ) thyroid imaging reporting and data systems ( T I‐RADS ) for thyroid nodules . Methods Cases of thyroid nodule who underwent surgery from January 2015 to M arch 2018 in 15 hospitals in Sichuan province were collected and the ultrasonographic features of thyroid nodules were retrospectively analyzed by trained senior ultrasound physicians using Kwak and ACR T I‐RADS classification methods . Totally ,12 712 thyroid nodules were observed ,7 023 thyroid nodules in 7 023 cases with complete ultrasound and surgical and pathological data were eventually enrolled in the study . T hyroid nodules with solid ,hypoechoic or very hypoechoic ,tall/wide ratio ≥ 1 , margin ill‐defined and microcalcification were classified as malignant signs of ultrasound . M alignant percentage was calculated and diagnostic tests were performed . Results ① T here was a statistical difference between the benign and malignant nodules in the two types of T I‐RADS classification ( P＜0 .01) . ② T he area under ROC curve of Kwak and ACR in the diagnosis of malignant nodules were 0 .89 and 0 .84 ,respectively . T he Youden index of Kwak and ACR were 0 .66 and 0 .57 ,respectively . ③Taking Kwak T I4B and ACR T R4 as critical points for malignancy ,the sensitivity ,specificity ,positive predictive value and negative predictive value of Kwak T I 4B were 75 .0% ,90 .9% ,83 .2% ,and 85 .9% , respectively . T he accuracy of Kwak T I4B was 84 .9% ; T he sensitivity ,specificity ,positive predictive value and negative predictive value of ACR T R4 were 88 .2% ,68 .9% ,62 .9% ,and 90 .8% ,respectively . T he accuracy of ACR T R4 was 76 .2% . T he Kappa value of Kwak TI4B and ACR T R4 was 0 .52 . T he χ2 value of Kwak T I4B and ACR T R4 was 2 174 .6 ( P ＜ 0 .01 ) . Conclusions T he diagnostic values of two T I‐RADS classification methods for thyroid malignant nodules are high . T he overall efficiency of Kwak T I‐RADS classification method is better than that of ACR TI‐RADS classification method .