OBJECTIVETo prepare and identify a polyclonal antibody (pAb) against (mouse) cysteinyl leukotriene receptor 1 (CysLT(1)) and to investigate the changes of CysLT(1) receptor expression in BV2 microglial cells after rotenone treatment.

METHODSRabbits were immunized with KLH-coupled CysLT(1) peptide to prepare the pAb. The titer of the pAb in rabbit plasma was detected by ELISA method, and the specificity of the pAb was tested by antigen blockade. After BV2 cells were treated with rotenone (0.01-1 μmol/L) for 24 h, the expression of CysLT(1) was determined by immunostaining, Western blotting and RT-PCR.

RESULTThe pAb showed a titer of 1/32728, and was not cross-reacted with antigens of CysLT(2) receptor and GPR17. Immunostaining, Western blotting and RT-PCR analysis showed the expression of CysLT(1) receptor in BV2 microglia. Rotenone at 1μmol/L significantly induced an increased expression of CysLT(1) receptor.

CONCLUSIONThe prepared CysLT(1) receptor polyclonal antibody has a high titer and high specificity to meet testing requirements of Western blotting and immunostaining; CysLT(1) is associated with rotenone-induced injury of BV2 microglial cells.

Animals ; Cells, Cultured ; Male ; Mice ; Microglia ; drug effects ; metabolism ; pathology ; Rabbits ; Receptors, Leukotriene ; immunology ; metabolism ; Rotenone ; pharmacology

BACKGROUNDBiofilms have given new insights to the understanding of pathogenesis of chronic rhinosinusitis (CRS). However, the link between biofilms formation and local inflammatory response remains poorly defined in CRS with nasal polys. The aim of this study was to determine the potential association of the presence of biofilms in the nasal mucosal tissues with clinical features in Chinese patients, which had CRS with nasal polyps (CRSwNP).

METHODSA total of 19 patients with CRSwNP and 12 patients with non-CRS were subjected to endoscopic surgery and their nasal mucosal tissue specimens were examined histologically and by scanning electron microscopy (SEM). Their demographic and clinical features were recorded.

RESULTSThirteen (68.4%) out of the 19 specimens from patients with CRSwNP, but none from control patients, were positive for biofilms that displayed typical characteristics of bacterial and fugal structures. The presence of biofilms in the nasal mucosal tissues was associated with significantly greater values of purulent nasal discharge and preoperative Lund-Kennedy scores, higher levels of serum total IgE and percentages of subjects with endoscopic surgery (ESS) history in patients with CRSwNP, and more severe inflammation in the nasal mucosal tissues of patients with CRSwNP.

CONCLUSIONOur study demonstrated the presence of biofilms in the nasal mucosal tissues of many patients, contributing to the understanding of the pathogenic process of CRSwNP in Chinese patients.

Adolescent ; Adult ; Biofilms ; Chronic Disease ; Female ; Humans ; Male ; Microscopy, Electron, Scanning ; Middle Aged ; Nasal Mucosa ; pathology ; Nasal Polyps ; etiology ; microbiology ; pathology ; Rhinitis ; etiology ; microbiology ; pathology ; Sinusitis ; etiology ; microbiology ; pathology

Adolescent ; Adult ; Aged ; Female ; Hepatitis B, Chronic ; pathology ; physiopathology ; Humans ; Liver ; pathology ; Liver Cirrhosis ; pathology ; Male ; Middle Aged ; Receptor, Cannabinoid, CB1 ; analysis ; Young Adult

OBJECTIVETo evaluate the effectiveness, safety and feasibility of endovascular angioplasty in treating long iliac artery chronic total occlusions (CTO).

METHODSThe clinical data from a consecutive series of patients with long (> 5 cm) iliac artery CTO who treated by endovascular angioplasty from January 2006 to December 2010 was retrospectively analyzed. There were 139 patients (157 limbs) with long iliac artery CTO treated by endovascular angioplasty in this study [male 93 and female 46, mean age (77 ± 10) years]. According to TASC II classification, there were 18 patients in type B, 89 patients in type C and 32 patients in type D. Recanalization of the occluded lesions was attempted with the left brachial and/or femoral access.

RESULTSThe ankle brachial index increased from 0.42 ± 0.19 before treatment to 0.81 ± 0.26 after treatment. The rate of technical success was 96.2% (151/157) and the patency rate of iliac artery was 94.1% (111/118) during the follow-up. Significant restenosis or reocclusion was found in 7 iliac lesions and there were no major interventional complications, such as iliac artery rupture, stent displacement, pseudoaneurysms, and arteriovenous fistula.

CONCLUSIONSEndovascular angioplasty is an effective, safe and feasible method in treating long iliac CTO with high patency rate. Combined left brachial and femoral access can increase the technical success rate significantly.

Aged ; Aged, 80 and over ; Angioplasty, Balloon ; methods ; Arterial Occlusive Diseases ; surgery ; Female ; Follow-Up Studies ; Humans ; Iliac Artery ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

This study was purposed to applicate the bone marrow indirect Coombs test and investigate its clinical significancies in diagnosis of immuno-related pancytopenia (IRP). 30 patients with pancytopenia including 22 cases of IRP and 8 cases of idiopathic cytopenia of undetermined significance (ICUS), and 15 patients with iron-deficiency anemia as controls were enrolled in this study. After incubation of the bone marrow supernatant of IRP patients and bone marrow nucleated cell (BMNC) of controls was used as experiment group, while the incubation of BMNC and bone marrow supernatant of controls was used as control group. After incubation for 45 min, the positive rate of membrane antibodies in bone marrow hematopoietic cells (CD15(+), GlyCoA(+) and CD34(+)cells) was detected by flow cytometry, and correlation analysis of positive rate with clinical data of patients were analyzed. The results showed that among 30 patients with pancytopenia (16 positive and 14 negative for bone marrow direct Coombs test) 16 cases showed positive for bone marrow indirect Coombs test, with positive rate 53.33. In the experiment group, the median positive rate of CD15(+)IgM was 0.34, which was significantly higher than that in control group (0.20, P < 0.05); the median positive rates of CD34(+) IgG and IgM were 0.64 and 0.21 respectively, which were significantly higher than those in control group (0.00, P < 0.05) and (0.00, P < 0.05); the positive rates of GlyCoA(+)IgG and IgM were (0.83 ± 0.75) and (2.12 ± 1.98) respectively, which were significantly higher than those in control group [(0.47 ± 0.43), P < 0.05, (0.68 ± 0.64), P < 0.01]; the positive rates of CD15(+) IgG and IgM were positively correlated with the ratio of CD5(+)B cells. The positive rates of GlyCoA(+) IgG and IgM negatively correlated with the Hb level, percentage of reticulocytes, the ratio of bone marrow erythroid lineage and DC1/DC2 positively correlated with the ratio of CD5(+)B cells and indirect bilirubin level. It is concluded that antibodies (IgG or IgM) aiming at the bone marrow hematopoietic cells exist in the supernatant of some IRP and ICUS patients, and may act on the membrane protein of the normal BMNC. These antibodies correlate with the prognosis of IRP.
Adolescent ; Adult ; Aged ; Bone Marrow Cells ; immunology ; Child ; Coombs Test ; Female ; Humans ; Male ; Middle Aged ; Pancytopenia ; diagnosis ; immunology ; Young Adult

OBJECTIVETo analyze the experience with diagnosis and surgical treatment of visceral artery aneurysms (VAAs).

METHODSFrom June 2003 to December 2008, 8 patients (2 male and 6 female) with 9 VAAs underwent surgical treatment. Mean age was 49 years (ranged from 30 to 72 years). The site of aneurysmal disease was splenic artery in 4 cases, superior mesenteric artery in 2 cases, renal artery in 2 cases (3 aneurysms). In 1 patient of splenic artery aneurysm, portal vein hypertension coexisted. All the VAAs of preoperative diagnostic workup consisted of a ultrasound, computed tomography (CT) scan, and digital subtraction angiography. Six patients were operated on and two patients was treated with endovascular procedures. Only one small VAAs was treated with follow-up.

RESULTSNo deaths or major complications occurred in the perioperative period. All the patients remained symptom free during a follow-up of 26.5 months (ranged from 2 to 60 months). Follow-up consisted of clinical and ultrasound scan examinations or CT scan at 1 and 6 months, and yearly thereafter.

CONCLUSIONSAggressive approach to the treatment of VAAs is essential. Elective open surgical treatment and an endovascular procedure of visceral artery aneurysms are both safe and effective, and offers satisfactory early and long term results. There is some evidence that small (< 2 cm) and asymptomatic VAAs may be safely observed.

Adult ; Aged ; Aneurysm ; surgery ; Blood Vessel Prosthesis Implantation ; Female ; Follow-Up Studies ; Humans ; Male ; Mesenteric Artery, Superior ; pathology ; surgery ; Middle Aged ; Renal Artery ; pathology ; surgery ; Retrospective Studies ; Splenic Artery ; pathology ; surgery ; Treatment Outcome

OBJECTIVETo investigate the expressions of STAT5 phosphorylation in CD34(+)CD38(-)CD123(+) bone marrow cells of the patients with myelodysplastic syndromes (MDS), and then evaluate the level of activation of STAT5 associated with cell proliferation in MDS clone cells.

METHODSThe bone marrow mononuclear cells (BMMNC) were extracted from 36 MDS patients and 14 normal controls. The mean fluorescence intensities (MFI) of phosphorylated STAT5(P-STAT5) in CD34(+)CD38(-)CD123(+) and CD34(+)CD38(-)CD123(-)cells, with or without the stimulation of 10 U/ml EPO, were examined by flow cytometry (FCM).

RESULTSWithout stimulation, the P-STAT5 MFI in CD34(+)CD38(-)CD123(+) cells of low/high risk MDS patients was 113.71 ± 67.22/173.05 ± 102.78, which was significantly higher than that of CD34(+)CD38(-)CD123(-) cells (58.84 ± 27.51/68.99 ± 50.42, P < 0.01, P < 0.05) and the normal controls CD34(+)CD38(-)CD123(-) cells (63.06 ± 21.06, P < 0.05), there was no significant difference between the CD34(+)CD38(-)CD123(-) cells of MDS patients and the normal control CD34(+)CD38(-)CD123(-) cells; With the EPO stimulation, the P-STAT5 MFI in CD34(+)CD38(-)CD123(+) cells of low/high risk MDS patients was 144.04 ± 58.11/239.45 ± 152.05, which was significantly higher than that of CD34(+)CD38(-)CD123(-) cells (68.41 ± 25, 10/64.21 ± 23.43, P < 0.01) and the normal controls CD34(+)CD38(-)CD123(-) cells (75.21 ± 27.02, P < 0.01), there was no significant difference between the CD34(+)CD38(-)CD123(-) cells of MDS patients and the normal control CD34(+)CD38(-)CD123(-) cells; The P-STAT5 MFI in the CD34(+)CD38(-)CD123(+) cells of low/high risk MDS patients with or without EPO stimulation were 21.80/28.86, which was significantly higher than that of CD34(+)CD38(-)CD123(-) cells (7.42/5.50, P < 0.01, P < 0.05) and the normal controls CD34(+)CD38(-)CD123(-) cells (6.39, P < 0.05), there was no significant difference between the CD34(+)CD38(-)CD123(-) cells of MDS patients and the normal controls CD34(+)CD38(-)CD123(-) cells; There was no significant difference of P-STAT5 MFI with or without EPO stimulation and the increased P-STAT5 MFI between the CD34(+)CD38(-)CD123(+) cells of low and high risk MDS.

CONCLUSIONSTAT5 associated with cell proliferation was activated in CD34(+)CD38(-)CD123(+) bone marrow cells in MDS, which had more significant reactions to EPO than CD34(+)CD38(-)CD123(-) cells, indicating that CD34(+)CD38(-)CD123(+) bone marrow cells might be the real malignant MDS clone cells in MDS.

Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; metabolism ; Bone Marrow Cells ; cytology ; metabolism ; Cell Proliferation ; Cells, Cultured ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; metabolism ; pathology ; Phosphorylation ; STAT5 Transcription Factor ; metabolism

Objective: To establish the quality control methods for the standard decoction of Puerariae Thomsonii Radix.Method:According to the preparation principles for traditional Chinese medicine (TCM) standard decoction,13 batches of Puerariae Thomsonii Radix from different origins were analyzed under the chromatography conditions established in this study and verified with methodology.By referring to Chinese Pharmacopoeia of 2015,puerarin was used as a quantitative indicator to calculate the transfer rate.In this study,the structures of main chromatographic peaks were also identified to clarify the main chemical constituents in the standard decoction.Result:The 13 batches of medicinal herbs were identified as Puerariae Thomsonii Radix,with a recovery rate of 98.0%,and RSD of 1.1%,indicating that the method was accurate and reliable.The transfer rate ranged from 41.4% to 60.0%,and the extraction rate was within the range of 15.7%-34.3%.The corresponding fingerprints were prepared for 13 batches of the standard decoction,and their similarities were all greater than 90.0%.The chemical constituents from Puerariae Thomsonii Radix were identified by mass spectrometry analysis,including citric acid,4'-O-glucosyl puerarin/daidzein-4',7-diglucoside,3'-hydroxy puerarin/genistein puerarin,puerarin apioside,daidzin puerossid A and daidzein,etc.Conclusion: The 13 batches of Puerariae Thomsonii Radix decoction in different origins had consistent properties with the basic properties of medicinal decoction pieces.The established method of quality evaluation can be used to systematically evaluate the standard decoction,providing reference for quality control of related decoction preparations of Puerariae Thomsonii Radix.

Objective To evaluate the application of adding ondansetron in morphine intravenous analgesia pump for prevention of postoperative nausea and vomiting(PONV)in women. Methods Data of surgical female inpatients who received general anesthesia and postoperative intravenous analgesia with morphine in our center from January 1st,2013 to December 31st,2014 were retrospectively analyzed. Based on whether ondansetron was added,patients were divided into the observation group(n=834) and the control group(n=856). Outcome variables including morphine consumption,visual analogue scale(VAS) score,nausea score,and total times of vomiting within 48 hours after surgery were collected and compared. Results Age(t=0.432,P=0.665),morphine consumption during the first post-operative day(t=0.599,P=0.548) and during the second post-operative day(t=0.236,P=0.813),maximum VAS score of postoperative pain during movement during the first post-operative day [3(2,4) vs. 3(2,5);Z=1.850,P=0.064] and at rest during the second post-operative day [0(0,1) vs. 0(0,1);Z=1.511,P=0.131] were not significantly different between two groups. While the maximum VAS score of postoperative pain at rest during the first post-operative day [0(0,2) vs.0(0,2);Z=2.435,P=0.015] and during movement during the second post-operative day [3(1,3)vs.3(2,4);Z=3.445,P=0.001] were significantly different between two groups. The maximum score of postoperative nausea(χ=9.810,P=0.020) and cumulative frequency of postoperative vomiting(Z=3.726,P=0.002)in the observation group were significantly lower than those in the control group during the first post-operative day;however,there was no significant difference during the second post-operative day(χ=5.017,P=0.170;Z=0.000,P=1.000). Logistic regression analysis showed that adding ondansetron in morphine intravenous analgesia pump was an independent influencing factor of reduced postoperative nausea. The probability of nausea during the first post-operative day was 0.781 time(P=0.015)of that in the control group and 0.736 time(P=0.030)during the second post-operative day. Conclusion Adding ondansetron in morphine intravenous analgesia pump may reduce PONV in women.

Cucurbitadienol has anti-inflammation, anti-cancer activities, and acts as a precursor of traditional Chinese medicine active ingredients mogroside and cucurbitacine. For construction of a Sacchromyces cerevisiae cell factory for production of cucurbitadienol, we firstly cloned a cucurbitadienol synthase (CBS) gene from Siraitia grosvenorii. Then, through heterologous expression of CBS in the triterpenoid chassis strain WD-2091, the engineered strain could produced 27.44 mg•L ⁻¹ cucurbitadienol, which was determined by GC-MS. Further regulation of CBS expression led to cucurbitadienol's titer increasing by 202.07% and reaching 82.89 mg•L ⁻¹ in the shake flask fermentation and 1 724.10 mg•L ⁻¹ in the high cell density fermentation. Our research promotes the cucurbitane-type tetracyclic triterpenoids synthesis pathway analysis progress and provides the basis for further obtaining cell factories for production of cucurbitadienol tetracyclic triterpenoids.