OBJECTIVETo observe the effect of CKJ Recipe (consisting of Cordyceps sinensis polysaccharide, amygdaloside, and gypenosides) containing serum on the activation of rat primary hepatic stellate cells (rHSCs) and to explore its pharmacological mechanism.

METHODSrHSCs were isolated form liver and cultured for four days. Then they were divided into the normal control group, the model group, and the CKJ group. rHSCs in the model group and the CKJ group were treated with 2.5 ng/mL transforming growth factor beta1 (TGF-beta1) in serum-free DMEM for 24 h. Serum free DMEM (containing no TGF-beta1) was taken as the control for the normal control group. rHSCs in the CKJ group were treated with 5% CKJ-containing serum for 24 h. rHSCs in the other two groups were treated with 5% blank serum for 24 h.The protein expression level of a smooth muscle actin (alpha-SMA) was determined using high throughput screening (HCS) and Western blot. mRNA expression levels of alpha-SMA, collagen I (Col-I), platelet-derived growth factor receptor beta (PDGF-betaR), TGF-beta1, transforming growth factor beta receptor 1 (TGF-betaR1), and transforming growth factor beta receptor 2 (TGF-beta R2) were detected using quantitative RT-PCR.

RESULTSCompared with the normal control group, the protein expression level of alpha-SMA, mRNA expression levels of alpha-SMA, Col-I, PDGF-betaR, TGF-beta1, TGF-betaR1, and TGF-betaR2 significantly increased in the model group (P<0.05, P<0.01). Compared with the model group, the protein expression level of alpha-SMA, mRNA expression levels of alpha-SMA, Col-I, PDGF-betaR, TGF-beta1, TGF-beta1, and TGF-beta R2 significantly decreased in the CKJ group (P<0.05, P<0.01).

CONCLUSIONCKJ containing serum could inhibit the protein expression level of o-SMA, which was probably related with inhibiting TGF-beta1 and its related receptors.

Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hepatic Stellate Cells ; metabolism ; Rats ; Transforming Growth Factor beta ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo investigate the diagnostic value of FICTION (Fluorescence Immunophenotyping and Interphase Cytogenetics as a Tool for the Investigation of Neoplasms) technique, combining immunofluorescence and fluorescence in situ hybridization (FISH), to detect genetic aberrations in multiple myeloma (MM).

METHODSBone marrow samples were collected from 18 MM and 2 plasma cell leukemia (PCL) patients. Probes targeting IgH and MMSET were prepared using a Nick Translation Kit from Bacterial artificial chromosome (BAC) clones. The immunophenotyping was achieved via the CD138 tyramide signal amplification (TSA)-mediated immunofluorescence, followed by FISH with the prepared probes \[t(4;14), t(11;14), t(14;16)\] and the commercial deletion probes (13q and p53) to detect common genetic aberrations in MM.

RESULTSAll the 20 samples were assayed with the probes mentioned above, and revealed 4 cases with t(4;14), 6 with t(11;14), 1 with t(14;16), 3 with p53 deletion; and 8 with 13q deletion. The remaining 4 cases had none of the 5 aberrations.

CONCLUSIONFICTION technique facilitates the detection of genetic abnormalities of MM in situ; enhances both efficiency and sensitivity of positive detection, thus, could be used as the screening test of molecular diagnosis of MM to guide coming-up risk-adapted therapy and evaluate prognosis.

Adult ; Aged ; Aged, 80 and over ; Cytogenetics ; Female ; Fluorescent Antibody Technique ; Humans ; Immunophenotyping ; In Situ Hybridization, Fluorescence ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; genetics

Objective To establish a method for quantitative determination of 4 chemical constituentsgeniposide,genipin-1-β-D-gentiobioside,arctiin and baicalin in the effective fraction of Zhibangreduping Formula.Methods The procedare of HPLC was performed on the chromatographic column of Diamonsil C_(18)(4.6 mm×150 mm,5μm),and the mobile phase in the gradient elution was acetonitrile-phosphoric acid solution(0.2%).The flow rate was 1.0 mL/min and the detection wavelength was 238 nm and 278 nm.Results The average recovery of geniposide WaS 99.02%(RSD=1.99%,n=6),genipin-1-β-D-gentiobioside,99.03%(RSD=2.83%,n=6),arctiin,101.12%(RSD=2.86%,n=6),and baicalin,100.05%(RsD=2.02%.n=6).Conclusion The results of determination in 3 batches of samples show that the method is easy and accurate,which can be used in the quantitative determination of 4 chemical constituents in the effective fraction of Zhibangreduping Formula.

BACKGROUNDNatural disasters have been frequent in recent years. Effective treatment of patients with cardiovascular disease following natural disasters is an unsolved problem. We aimed to develop a novel miniature mobile cardiac catheterization laboratory (Mini Mobile Cath Lab) to provide emergency interventional services for patients with critical cardiovascular disease following natural disasters. A feasibility study was performed by testing the Mini Mobile Cath Lab on dogs with ST-elevation myocardial infarction (STEMI) model in a hypothetical natural-disaster-stricken area.

METHODSThe Mini Mobile Cath Lab was transported to the hypothetical natural-disaster-stricken area by truck. Coronary angiography and primary percutaneous coronary intervention (PCI) were performed on six dogs with STEMI model. The transportation and transformation of the Mini Mobile Cath Lab were monitored and its functioning was evaluated through the results of animal experiments.

RESULTSThe Mini Mobile Cath Lab could be transported by truck at an average speed of 80 km/h on mountain roads during daytime in the winter, under conditions of light snow (-15°C to -20°C/-68°F to -59°F). The average time required to prepare the Mini Mobile Cath Lab after transportation, in a wetland area, was 30 minutes. Coronary angiography, and primary PCI were performed successfully.

CONCLUSIONThis preliminary feasibility study of the use of the Mini Mobile Cath Lab for emergency interventional treatment of dogs with STEMI indicated that it may perform well in the rescue of critical cardiovascular disease following natural disasters.

Angioplasty, Balloon, Coronary ; Animals ; Cardiac Catheterization ; Cardiovascular Diseases ; therapy ; Coronary Angiography ; Disasters ; Dogs ; Electrocardiography ; Feasibility Studies ; Laboratories ; Myocardial Infarction ; therapy

Objective To investigate the prevalence and genotype distribution of human papillomavirus (HPV) in the region of Guangxi.Methods We retrospectively analyzed 15774 individuals from the Outpatient and inpatient Unit as well as Physical Examination Departments of the People's Hospital of Guangxi Zhuang Autonomous Region between May 2010 and March 2017.Exfoliated c ellsor swab specimens were collected,and the genotypes of HPV were determined by Polymerase Chain Reaction (PCR) and reverse blot assay.Results The prevalence of HPV infection was 38.54％ (6080/15774).The infection rate of single genotype and multiple genotypes were 25.60％ (4038/15774) and 12.95％ (2042/15774),respectively.In single infection patterns,the most common genotypes included HPV 6 (10.54％,1663/15774),52 (3.91％,616/ 15774),16 (2.16％,340/15774),11 (2.14％,338/15774),and 58 (2.00％,316/15774).While among the multiple infections patterns,HPV 52+53 (4.26％,87/2042) and 52+58 (3.23％,66/ 2042) were common,and followed by HPV 16+43 (2.40％,49/2042),16+52(2.30％,47/2042),6+42 (2.15％,44/2042) and 6+43 (2.15％,45/2042).HPV 52,16,58,51,53 and 18 were the top six high risk (HR) genotypes of HPV,accounting for 26.77％ (4222/15774,95％ CI =26.08-27.46);while HPV 6,11 and 43 were the leading low-risk (LR) genotypes of HPV,accounting for 27.77％ (4380/15774,95％CI =27.07-28.47).The overall ratio of single infection to multiple infections was 1.98 (4038 vs.2042).Conclusion HPV 6,52,16,11,58,18 were the main HPV infection genotypes,and 52+53 and 52+58 were common infection combinations in Guangxi Zhuang Autonomous region.The HPV multiple infections were increased.Apparently,more HPV low risk genotypes were seen in male patients that should be aware.

OBJECTIVETo establish xenotransplated mouse model by non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice with primary myeloma cells.

METHODSThe model of xenograft was established in NOD/SCID mice by tail vein injection of mononuclear cells from two end stage multiple myeloma patients, three mice were inoculated for each patient. Mice were monitored weekly for body weight. Two weeks later, the human CD45(+) cells from peripheral blood of mice were evaluated by flow cytometry (FCM). The experiment endpoint was body weight loss up to 20% or had pale, vertical hair and listlessness, then spleen and liver were studied by histologic analysis, the human CD45(+)CD38(+) cells from spleen, lymph node, peripheral blood and bone marrow were evaluated by FCM.

RESULTSBody weight of mice in group patient 1 and group patient 2 decreased seven and five weeks after inoculation respectively; the human CD45(+)CD38(+) cells appeared in the peripheral blood (26 ± 4) and (16 ± 4) days after inoculation in group patient 1 and group patient 2 respectively, and increased by time, reaching (16.2 ± 3.0)% and (31.3 ± 3.5)%, respectively at the endpoint; the spleen, liver and lymph node of both groups enlarged, the typical malignant plasma cells were observed in them. The human CD45(+)CD38(+) cells were detected in spleen, lymph node and bone marrow by FCM.

CONCLUSIONOur study successfully established a NOD/SCID mouse model xenotransplated with human primary myeloma cells.

Aged, 80 and over ; Animals ; Cell Line, Tumor ; Disease Models, Animal ; Humans ; Male ; Mice ; Mice, Inbred NOD ; Mice, Nude ; Mice, SCID ; Middle Aged ; Multiple Myeloma ; Neoplasm Transplantation

The efficacy of rhIL-11 in treating thrombocytopenia and neutropenia in gamma-irradiated rhesus monkeys and the variation in curative effect due to difference of administration times were studied. Healthy rhesus monkeys were exposed to 3.0 Gy (60)Co total body irradiation (TBI) to result in pancytopenia for three weeks. Treatment with rhIL-11 (30, 60 or 120 micro g.kg(-1).day(-1)) on early days (days 0 - 13 after TBI) could significantly improve the nadir of platelet count. Although the nadir of leukocyte count was not improved, the duration below 50% of its baseline value was shortened similarly to that of platelet. During the first two weeks after TBI, erythrocyte numbers of the animals treated with these doses of rhIL-11 were lower than those of the control group at first but they became higher beginning from the third week. Four monkeys were treated with rhIL-11 at 60 micro g.kg(-1).day(-1) on days 13 - 26 after TBI. The numbers of their peripheral blood cells followed the similar decrease patterns as those of control group during the first three weeks, then they were improved rapidly. By semi-solid bone marrow cell culture it was demonstrated that rhIL-11 could stimulate bone marrow cells to form more CFU-Meg, CFU-Mix, CFU-E, BFU-E and CFU-GM in vitro. Histopathological observation revealed that bone marrow of the control group was devoid of hematopoietic cells and bleeding, being contrary to that of the group treated with rhIL-11, in which the cells proliferated actively. The results suggest that rhIL-11 can accelerate hematopoietic recovery of irradiated monkeys.

OBJECTIVETo investigate the diagnosis and surgical treatment of adult primary retroperitoneal malignant tumor (APRMT).

METHODSThe clinical data of 98 cases with APRMT underwent resection from January 1990 to April 2003 were analyzed retrospectively.

RESULTSAmong the 98 cases, complete excision were performed in 79 cases (80.6%), palliative excision in 16 cases (16.3%), tumor biopsy only in 3 cases (3.1%). Resection of involved adjacent organs were carried out in 25 cases (25.5%) and the re-operation rate for recurrence was 28.6% (28 cases). The 1, 3, 5 year survival rates for 79 cases with complete resection were 93.7%, 73.4% and 34.2%, respectively. The 1, 3, 5 year survival rate for 16 cases with palliative resection were 75.0%, 6.3% and 6.3%, respectively.

CONCLUSIONSCertain imaging examinations are crucial to the diagnosis and preoperative evaluation of APRMT. Resection of the involved organs could improve resection rate and prognosis. For the recurrent cases, earlier reoperation is strongly recommended.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Prognosis ; Retroperitoneal Neoplasms ; diagnosis ; surgery ; Retrospective Studies ; Survival Analysis ; Treatment Outcome

BACKGROUNDBivalirudin was widely used as an anticoagulant during coronary interventional procedure in western countries. However, it was not available in China before this clinical trial was designed. This randomized, single-blind and multicenter clinical trial aimed to evaluate the efficacy and the safety of domestic bivalirudin during percutaneous coronary intervention (PCI).

METHODSA randomized, single-blind, multicenter trial was designed. Elective PCI candidates in five centers were randomized into a bivalirudin group and a heparin group, which were treated with domestic bivalirudin and non-fractional heparin during the PCI procedure. The efficacy was evaluated by comparing the activated coagulation time (ACT), the procedural success rate (residual stenosis < 20% in target lesions without any coronary artery related adverse events within 24 hours after PCI), and the survival rate without major adverse cardiac events at 30 days after PCI between the two groups. Safety was evaluated by the major/minor bleeding rate.

RESULTSA total of 218 elective PCI patients were randomized into a bivalirudin group (n = 110) and heparin group (n = 108). Except for two patients needing additional dosing in the heparin group, the ACT values of all other patients in both groups were longer than 225 seconds at 5 minutes after the first intravenous bolus. Procedural success rates were respectively 100.0% and 98.2% in the bivalirudin group and heparin group (P > 0.05). Survival rates without major adverse cardiac events at 30 days after PCI were 100.0% in the bivalirudin group and 98.2% in the heparin group (P > 0.05). Mild bleeding rates were 0.9% and 6.9% (P < 0.05) at 24 hours, and 1.9% and 8.8% (P < 0.05) at 30 days after PCI in the bivalirudin group and heparin group respectively. There was one severe gastrointestinal bleeding case in the heparin group.

CONCLUSIONSDomestic bivalirudin is an effective and safe anticoagulant during elective PCI procedures. The efficacy is not inferior to heparin, but the safety is superior to heparin.

Aged ; Antithrombins ; adverse effects ; therapeutic use ; Female ; Heparin ; therapeutic use ; Hirudins ; adverse effects ; Humans ; Male ; Middle Aged ; Peptide Fragments ; adverse effects ; therapeutic use ; Percutaneous Coronary Intervention ; Recombinant Proteins ; adverse effects ; therapeutic use ; Single-Blind Method ; Survival Rate ; Whole Blood Coagulation Time

Objective:The hydrogen sulfide (H2 S) role in pathogenesis of various diseases were wildly addressed in recent decade.The circulatory (plasma or serum) and biological fluid H2S measurement is still an enormous issues due to the technical limitation.This paper aimed to develop a novel measurement method based on fluorescence probe.Methods:Firstly,20 μL ethanol was used to dissolve 100 pmol fluorescence probe,then added in a 96-well plate.An equal volume of ethanol was also added to the blank well of the plate.The plate was placed in a dark room for about 1 h until the fluorescence probe was evenly coated in the 96-well microplate and dried.The plate was frozen at-20 ℃ for later use.Secondly,the plasma or serum sample was added with saturated ammonium sulfate buffer (pH 7.8) and then centrifuged to remove the proteins.The equal volume supernatant liquid was added to the probecoated well and the probe-uncoated well.The plate was incubated in a dark environment at 37 ℃ for 2 h.Finally,after incubation,the fluorescence density was acquired at λEx/λEm 340/445 nm in a microplate reader.The differences of the fluorescence density values between the probe-coated well and probeuncoated well were counted and H2S concentration of plasma/serum was calculated by standard curve with NaHiS.Results:The method had high sensitivity (from 0.3 to 100 μmol/L) and specificity for measuring H2S as compared with other biologically relevant reactive sulfur species and sulfur-containing amino acid.Serum H2S concentrations were assayed in 188 health volunteers using this method [(12.1 ±3.5) μmol/L,95％ CI:4.6-19.8 μmol/L],and the frequency distribution showed a normal tendency(one-sample Kolmogorov-Smirnov test,P ＞ 0.1).The serum H2S concentrations in 30 hypertension patients were decreased compared with 22 age-and gender-matched health individuals (paired-samples t test,t =9.937,P ＜ 0.001).There were no differences of H2S concentration in serum [(19.66 ±2.32) μmol/L] or plasma [(18.67 ±2.07) μmol/L],between the samples acquired from artery [(19.34 ±0.51) μmol/L] or vein [(18.99 ±0.50) μ mol/L] of male Wistar rats (repeated measurement of ANOVA,P =0.38).One week frozen samples did not affect the detection.The values of the repeated measurement did not differ (two-way ANOVA,P ＞ 0.05).Conclusion:The present method is easily performed with high sensitivity,specificity and repeatability for circulatory H2S.It is also quick and may apply for large samples.