To optimize the extraction technology of the total saponins in Dipsaci Radix, the extraction efficiency of total saponins was investigated with respecting to three variables including time, ethanol concentration and liquid-to-solid ratio. On the basis of a series of one-factor-at-a-time experiments, a polynomial regression model equation was fitted by the combined use of central composite experimental design and regression analysis. By analyzing the regression model using response surface analysis, the optimum extraction conditions of total saponins from Dipsaci Radix were identified as follows: 7 times of 55% alcohol, refluxing 3 times, 2.8 hours every time, and the extraction efficiency of total saponins was up to 0.201 1 g·g-1 and the asperosaponin Ⅵ was up to 0.015 4 g·g-1 under the optimized conditions. Confirmatory experiment indicated the good prediction ability of the established model and provided the basis for further development and utilization of Dipsaci Radix.

BACKGROUNDIn qualitative diagnosis of bile duct stenosis, single diagnostic measure is difficult to make a correct diagnosis, to combine several diagnostic techniques may be helpful to make an accurate diagnosis. The aim of this study was to evaluate the value of intraductal ultrasonography (IDUS), endoscopic brush cytology and K-ras, P53 gene mutation in the early diagnosis of malignant biliary stricture.

METHODSFrom February 2012 to February 2013, 84 patients with suspected malignant biliary stricture were performed IDUS firstly, then endoscopic brush cytology and finally K-ras, P53 gene mutation detection, the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of all above ways were evaluated and compared.

RESULTSOf 84 patients, 52 cases were ultimately diagnosed malignant biliary stenosis; of which, 9 cases had no recurrence or metastasis to other organs after radical operation during the follow-up period. IDUS combined with brush cytology and K-ras + P53 gene mutation detection had obvious advantage in the sensitivity, accuracy and negative predictive value than any other joint detection and single detection (the advantage was more significant compared with IDUS + brush cytology or any single detection P < 0.01). There were obvious statistical significance in the sensitivity and accuracy between IDUS + brush cytology + P53 or IDUS + brush cytology + K-ras and IDUS + brush cytology or IDUS (P < 0.05). There was no statistical significance in the sensitivity, specificity, positive predictive value, negative predictive value and accuracy between IDUS + brush cytology + P53 and IDUS + brush cytology + K-ras (P > 0.05).

CONCLUSIONSIDUS combined with brush cytology and K-ras, P53 gene mutation detection is better than the separate detection and contribute to the early diagnosis of malignant biliary stricture. Its more widespread use is recommended.

Aged ; Aged, 80 and over ; Bile Duct Diseases ; diagnosis ; genetics ; Bile Duct Neoplasms ; diagnosis ; genetics ; Bile Ducts ; pathology ; Constriction, Pathologic ; diagnosis ; genetics ; Female ; Genes, p53 ; genetics ; Genes, ras ; genetics ; Humans ; Male ; Middle Aged ; Mutation

Objective To analyze miR-92a expression and its clinical significance in the plasma of systemic lupus erythematosus (SLE) patients.Methods Plasma samples from 44 SLE patients,16 rheumatoid arthritis (RA) patients and 20 healthy controls were collected.The small RNAs in these plasma samples were isolated and reversely transcribed.Using cel-miR-39 as the external reference,the levels of miR-92a expression were detected by real-time polymerase chain reaction (PCR) method.MiR-92a and cel-miR-39 were analyzed by real-time fluorescence quantitative PCR and agarose gel electrophoresis.The sensitivity and specificity of miR-92a as SLE were analyzed by receiver-operating characteristic (ROC) curve.The correlation between the levels of miR-92a expression and the clinic pathological features of SLE and biological significance of miR-92a expression in SLE were further analyzed by Pearson or Chi-square test.Results Our data indicated that the plasma levels of miR-92a expression was 49.20 (5.33,95.17) in SLE patients,411.30 (320.84,504.69) in healthy controls,and 25.59(11.20,30.54) in RA patients.The difference was significant (x2=40.77,P＜0.01).The area under the ROC curve (AUC) was 0.958 for discriminating between SLE patients and normal subjects and 1.00 for discriminating between RA patients and healthy controls.The levels of miR92a expression cutoff values were set the as 198.59 for healthy control and 85.35 for RA patients,the diagnostic sensitivity and specificity were 93.2％,90％,and 100％,100％,res-pectively.The analysis of the correlation between miR-92a expression and the clinic pathological features of SLE had shown that the levels of plasma miR-92a expressions were much lower in SLE patients with down-regulated complement C3,and up-regulated urea nitrogen,creatinine,LDH,ATH (all P＜0 05).Conclusion Down-regulated miR-92a expression in plasma of SLE may be involved in the SLE disease occurrence or development and could be used as a novel potential diagnostic biomarker for SLE.

OBJECTIVETo investigate whether analgesic effect of electroacupuncture (EA) is affected by p38 mitogen-activated protein kinase (p38 MAPK) on microglia.

METHODSThere were two experiments. The experiment 1: 40 male Sprague-Dawley (SD) rats were randomly divided into the normal, surgery, EA and sham EA groups, and the L5 spinal nerve ligation (SNL) on the right side was used to establish neuropathic pain model. EA was applied to bilateral Zusanli (ST36) and Kunlun (BL60) at 24, 48 and 72 h after SNL for 30 min, once per day. The paw withdrawal thresholds (PWTs) were measured before surgery (as base) and at 24, 25, 49 and 73 h after surgery. Phospho-p38 MAPK (p-p38 MAPK), oxycocin-42 (OX-42, marker of microglia), and glial fibrillary acidic protein (GFAP, marker of astrocyte) in bilateral spinal cord dorsal horn (SCDH) were detected by immunofluorescence, respectively. The experiment 2: 40 male SD rats were cannulated for SNL-induced neuropathic pain, and then were randomly divided into the dimethyl sulfoxide (DMSO), EA plus DMSO, 4-(4-fluorophenyl)-2-(4-methylsulfonylpheny)-5-(4-pyridyl)-1H-imidazole (SB203580) and EA plus SB203580 groups. SB203580 (30 nmol/L) was administered 5 min prior to EA treatment. The PWTs and OX-42 in bilateral SCDH were measured as mentioned above.

RESULTSSNL-induced neuropathic pain reduced PWTs and increased the expression of p-p38 MAPK and OX-42 in bilateral lumbar SCDH of rats (P<0.01). Spinal p-p38 MAPK was only co-localized with OX-42 in our study. EA treatment significantly alleviated SNL-mediated mechanical hyperalgesia, and suppressed the expression of p-p38 MAPK and OX-42 in lumbar SCDH (P<0.05 or P<0.01). Intrathecal injection of low dose SB203580 had no influence on PWTs (P>0.05), but significantly inhibited the expression of OX-42 positive cells in bilateral SCDH (P<0.01 or P<0.05). EA plus SB203580 synergistically increased PWTs, and reduced the expression of bilateral spinal OX-42 (P<0.01 or P<0.05).

CONCLUSIONSThe central mechanism of EA-induced anti-hyperalgesia may be partially associated with the reduced expression of p-p38 MAPK, and subsequently reducing the activation of OX-42 in neuropathic pain. Therefore, EA may be a new complementary and alternative therapy for neuropathic pain.

Animals ; Biomarkers ; metabolism ; CD11b Antigen ; metabolism ; Electroacupuncture ; Fluorescent Antibody Technique ; Hyperalgesia ; pathology ; therapy ; Imidazoles ; pharmacology ; Ligation ; Male ; Microglia ; drug effects ; enzymology ; pathology ; Neuroglia ; drug effects ; metabolism ; Phosphorylation ; drug effects ; Posterior Horn Cells ; drug effects ; enzymology ; pathology ; Pyridines ; pharmacology ; Rats, Sprague-Dawley ; Spinal Nerves ; drug effects ; pathology ; p38 Mitogen-Activated Protein Kinases ; metabolism

【Objective】 To study the effect of intravenous immunoglobulin(IVIG) on the detection of blood transfusion compatibility in patients. 【Methods】 56 patients, submitted to our Hospital from March 1, 2017 to December 31, 2020, were enrolled as the research objects. They had negative unexpected antibody screening, major crossmatch incompatibility with the same blood type donors, and had a history of IVIG infusion. ABO and RhD blood groups typing, unexpected antibodies screening, crossmatch, direct antiglobulin test, indirect antiglobulin test, and acid elution test were all conducted by microcolumn gel method. 【Results】 After IVIG infusion, the initially major crossmatch incompatibility with the same blood type donors turned into compatiblity with O-type donors. Among them, 2 patients had transient discrepancy in ABO forward and reverse blood typing due to the IVIG infusion. IgG anti-A were detected in the red blood cell elution of 37 A-type patients; IgG anti-B in 2 B-type patients; 3 cases of IgG anti-A+ anti-B and 14 cases of solo IgG anti-A in 17 AB-type patients. 3 batches of IVIG preparations were detected randomly, IgG anti-A titer was 32-64, and IgG anti-B titer was 8-16. 【Conclusion】 The discrepancy in ABO forward and reverse blood typing and major crossmatch incompatibility with the same blood type donors may occur after non-O type patients received IVIG, which contains IgG types of anti-A and anti-B. In this situation, it is recommended to prepare major crossmatched O-type washed red blood cells to ensure the safety and effectiveness of clinical blood transfusion.

【Objective】 To explore the influencing factors of perioperative red blood cell transfusion in patients underwent lung transplantation, so as to provide reference for perioperative blood management (PBM) of lung transplantation patients. 【Methods】 The clinical data of 173 lung transplant patients completed in China-Japan Friendship Hospital from March 2017 to June 2019 were retrospectively analyzed. The patients were divided into two groups according to perioperative red blood cell transfusion volume: large blood transfusion group (transfusion red blood cell volume ≥6 U, n=66) and non-large blood transfusion group (red blood cell transfusion volume <6 U, n=107). The basic information, preoperative laboratory test results, and surgical status of the two groups were statistically analyzed.The clinical data of the two groups were analyzed by univariate analysis. The factors of P<0.15 were included in the binary logistic regression analysis, and the independent influencing factors of perioperative massive blood transfusion in patients with lung transplantation were found. 【Results】 Univariate analysis of clinical data of the two groups of patients (large blood transfusion group vs. non-large blood transfusion group) showed that the differences of smoking history ratio [44(66.7%) vs 87(81.3%)], BMI(20.8±4.5 vs 22.5±4.0)(P<0.05), preoperative Hb [124(111, 138.8) vs 138(126, 149)], preoperative Hct [37.9(34.8, 42.5) vs 41.3(37.9, 44.6)], surgery duration(327.9±107.7 vs 238.4±77.0), intraoperative blood loss(1 108.6±1342.0 vs 341.8±270.8) and single lung transplantation [28(42.4%) vs 84(78.5%)] (P<0.01) were statistically significant. Logistic regression analysis showed that intraoperative blood loss (OR=1.001, P<0.05), surgery duration (OR=1.006, P<0.05), preoperative Hb (OR=0.973, P<0.01), lung transplantation type(single or double lung transplantation)( OR=0.247, P<0.05) and extracorporeal membrane oxygenation (ECMO) (OR=0.187, P<0.01) were independent factors influencing red blood cell transfusion during lung transplantation. 【Conclusion】 Intraoperative blood loss and surgery duration are risk factors for massive blood transfusion during the perioperative period. And the use of ECMO, preoperative Hb, single lung transplantation (compared to double lung transplantation) are protective factors for perioperative massive blood transfusion.

Using chemoproteomic techniques, we first identified EIF2AK2, eEF1A1, PRDX3 and VPS4B as direct targets of berberine (BBR) for its synergistically anti-inflammatory effects. Of them, BBR has the strongest affinity with EIF2AK2 via two ionic bonds, and regulates several key inflammatory pathways through EIF2AK2, indicating the dominant role of EIF2AK2. Also, BBR could subtly inhibit the dimerization of EIF2AK2, rather than its enzyme activity, to selectively modulate its downstream pathways including JNK, NF-κB, AKT and NLRP3, with an advantage of good safety profile. In EIF2AK2 gene knockdown mice, the inhibitory IL-1β, IL-6, IL-18 and TNF-α secretion of BBR was obviously attenuated, confirming an EIF2AK2-dependent anti-inflammatory efficacy. The results highlight the BBR's network mechanism on anti-inflammatory effects in which EIF2AK2 is a key target, and inhibition of EIF2AK2 dimerization has a potential to be a therapeutic strategy against inflammation-related disorders.

Objective: Comparing the benefit of Abidor, lopinavir/ritonavir and recombinant interferon α-2b triple combination antiviral therapy and lopinavir/ritonavir and interferon dual combination antiviral therapy to hospitalized novel coronavirus pneumonia 2019 in Zhejiang province. Methods: A multi-center prospective study was carried out to compare the effect of triple combination antiviral therapy with dual combination antiviral therapy in 15 medical institutions of Zhejiang Province. All patients were treated with recombinant interferon α-2b (5 million U, 2 times/d) aerosol inhalation. 196 patients were treated with abidol (200 mg, 3 times/d) + lopinavir / ritonavir (2 tablets, 1 time/12 h) as the triple combination antiviral treatment group. 41 patients were treated with lopinavir / ritonavir (2 tablets, 1 time/12 h) as the dual combination antiviral treatment group. The patients who received triple combination antiviral therapy were divided into three groups: within 48 hours, 3-5 days and > 5 days after the symptom onset. To explore the therapeutic effects of triple combination antiviral drugs and dual combination antiviral drugs, as well as triple combination antiviral drugs with different antiviral initiate time. SPSS17.0 software was used to analyze the data. Results: The time of virus nucleic acid turning negative was (12.2 ± 4.7) days in the triple combination antiviral drug group, which was shorter than that in the dual combination antiviral drug group [(15.0 ± 5.0) days] (t = 6.159, P < 0.01 ). The length of hospital stay [12 (9, 17) d] in the triple combination antiviral drug group was also shorter than that in the dual combination antiviral drug group [15 (10, 18) d] (H = 2.073, P < 0.05). Comparing the antiviral treatment which was started within 48 hours, 3-5 days and > 5 days after the symptom onset of triple combination antiviral drug group, the time from the symptom onset to the negative of viral shedding was 13 (10,16.8), 17 (13,22) and 21 (18-24) days respectively (Z = 32.983, P < 0.01), and the time from antiviral therapy to the negative of viral shedding was (11.8±3.9) , (13.5±5.1) and (11.2±4.3) d. The differences among the three groups were statistically significant (Z=32.983 and 6.722, P<0.01 or<0.05). Conclusions The triple combination antiviral therapy of Abidor, Lopinavir/Litonavir and recombinant interferon α-2b showed shorter viral shedding time and hospitalization time compared with the dual combination antiviral therapy. The earlier the time to initiate triple antiviral treatment, the shorter the time of virus shedding.