Objective To evaluate if the assay of platelet-associated immunoglobulin G (PAIgG) can improve the sensitivity of monoclonal antibody immobilization of platelet antigens assay (MAIPA) in immune thrombocytopenic purpura (ITP).Methods Anti-GP Ⅱb/Ⅲa and anti-GP Ⅰb/Ⅸ autoantibodies were detected by a modified MAIPA;PAIgG was detected by competitive ELISA.MAIPA and PAIgG were done simultaneously in 190 patients with thrombocytopenia,in which 132 were diagnosed as ITP according to the diagnostic criteria Kappa test was used to analyze the concordance between MAIPA and PAIgG.Results The sensitivities of PAIgG alone or in parallel with MAIPA to ITP were 32.6% and 37.2% higher than MAIPA alone respectively,while their specifieities were 51.7% and 56.9% lower respectively.The sensitivity,specificity,positive predictive value,negative predictive value of PAIgG in series with MAIPA were all not superior to MAIPA alone.The kappa values between MAIPA and PAIgG in ITP and non-immune thrombocytopenia were 0.129 and 0.012 respectively,which meant the concordance between them was poor.Conclusion The detection of PAIgG should not be used as screening before MAIPA in distinguishing immune from non-immune thrombocytopenia.

Background:Small intestinal bleeding is difficult to diagnose and treat because of its complex etiology and limit to examination method. Aims:To analyze the etiology,diagnosis,treatment and prognosis of small intestinal bleeding. Methods:The clinical data of 118 consecutive patients with small intestinal bleeding admitted from Oct. 2006 to Oct. 2016 at Daping Hospital,the Third Military Medical University were retrospectively analyzed. Results:Melena was the most common manifestation of small intestinal bleeding (41. 5％),followed by dark bloody stool,positive fecal occult blood test,hematochezia,and anemia with unknown cause. The major causes of bleeding were benign or malignant tumors (43. 2％),vascular lesions (28. 0％)and inflammatory lesions (15. 3％). Diagnosis was made by means of capsule endoscopy,colonoscopy,digital subtraction angiography (DSA),barium meal examination,multi-slice CT (MSCT)and CT enterography (CTE). Forty-one patients were treated by surgical operation,7 by selective arterial embolization,2 by endoscopic therapy,56 by conservative therapy,and all these patients achieved hemostasis. One patient died of massive hemorrhage and 11 were discharged with giving up of treatment. Conclusions:The leading cause of small intestinal bleeding is tumor,followed by vascular and inflammatory lesions. Capsule endoscopy is able to make definite diagnosis with high accuracy,and MSCT is the most widely used diagnostic approach. In addition to conventional treatment,surgical operation,interventional and endoscopic therapies also play important roles in treating small intestinal bleeding.

Objective To investigate the immunomodulatory effect of mesenchymal stem cells (MSCs) and their role in prolonging allograft survival in rat heart transplantation. Methods Inbred Wistar rats were used as donors, and Fisher 344 as recipients. MSC were isolated from femur and tibia bone marrow of donors and cultured in vitro. Mixed lymphocyte reaction assays were performed to assess the immunosuppressive effects of different concentrations of MSC on allogeneic T cell proliferation. Cardiac allograft model was established and according to different intervention measures recipients were divided into two groups (MSC treatment group and control group) (n=8 in each group). In MSC treatment group, recipients were infused with 2×106 MSC via the tail vein at designated intervals (one week before operation, during operation and consecutive three days postoperation), while in control group, the recipients were treated with Ringer's solution at the same interval& At day 5 posttransplantation real-time PCR was used to detect the changes in the expression of Thl and Th2 cytokine genes in transplanted hearts. Results In vitro allogeneic T cell response was greatly suppressed by MSC in a dose-dependent manner. Real-time PCR revealed that IL-1β,IFN-γ, IL-4 and IL-10 were expressed in MSC treatment group, while IL-4 and IL-10 were not expressed in control group but with significantly higher expression of IL-1β and IFN-γ. As compared with control group, survival of MSC-treated allografts was markedly prolonged as compared with control group (mean survivaldays: 12.4±5.3 vs 6.4±2.0, P＜0.05). Conclusion Intravenous adrninistmtion of MSC can prolong the survival of transplanted heart possibly by induction of allograft tolerance through changing Th1/Th2 balance.

Objective To investigate the possible role of interstitial cell of Cajal (ICE)in the pathogenesis of diabetic gastroparesis and the protective effect of"Xiaopi prescription (消痞方)".Methods Fifty healthy male Spregue-Dawley (SD) rats were randomly divided into 5 groups (each n=10):normal, model,and 3 Xiaopi prescription groups:low,middle and high dosages.Diabetes was induced by intravenous injection of alloxan,and equal amount of normal saline was intravenously injected in the normal group.Gastric lavage method was used to administer the traditional Chinese medicine decoction of Xiaopi prescription in corresponding amount (5,10 and 20 g?kg~(-1)?d~(-1)) in respective low,middle and high dosage groups.In the normal control group and diabetic model group,only equal amount of normal saline was administered into the stomach.Gastric emptying rate was measured by method of nutritious semisolid paste;c-kit positive cells of ICC were quantitatively measured with immunohistochemistry assay and computer image analysis system;c-kit mRNA positive cells were quantitatively measured with in situ hybridization and computer image analysis system.Results①Gastric emptying rate:The rate was significantly lower in the model group than that in the normal control group (P0.05),but higher than those in the model group and the low dosage group (all P0.05).②c-kit immmunohistochemistry:c-kit positive cell presented yellow in color,and its membrane was stained yellow,this kind of cells primarily were distributed around the neural plexus in the inter-space between the circular and longitudinal muscular fibers, and around the ganglionic cells forming"sheath-like"structure.The results of numbers of c-kit positive cells in the various groups:the number of the cells in the model group was significantly lower than that in the normal group (P0.05),but the numbers in the former two dosage groups were obviously higher than those in the model group (all P0.05),being significantly lower than that in the normal group,middle and high dosage groups (all P0.05),but obviously higher than those in the model group (all P0.05),being significantly lower than that in the normal,middle and high dosage groups (all P

BACKGROUND:Gemcitabine hydrochloride is a water-soluble anticancer drug that induces apoptosis in tumor cells, but it has an excessive release in vivo. OBJECTIVE:To evaluate the effect of poly(lactic acid) (PLA) electrospun fiber membranes carrying gemcitabine hydrochloride on the growth of human osteosarcoma cell lines MG-63. METHODS:PLA electrospun fiber members with (experimental) or without (control) gemcitabine hydrochloride were fabricated and characterized. Two kinds of fiber membranes were immersed in low-glucose DMEM medium, and the supernatants were collected in the two groups at 3, 5, 7 days, respectively. Passage 5 human osteosarcoma cell lines MG-63 were inoculated into 96-well plates containing low-glucose DEME with 15％ fetal bovine serum, and divided into seven groups. Groups 1-3 were cultured in the experimental supernatants of 3, 5, 7 culture days, and groups 4-6 were cultured in the control supernatants of 3, 5, 7 culture days, respectively. The remaining group acted as the negative control with no supernatant. Thereafter, cell counting kit-8 was used to detect cell proliferation, and RT-PCR was used to measure expression of Bcl-2 and Bax at 3 days of culture. RESULTS AND CONCLUSION:(1) No obvious particle was found on the smooth and even surface of the fiber members in the experimental and control groups. There was no significant difference in fiber diameter, contact angle and tensile strength between the two kinds of fiber membranes. (2) The results of cell counting kit-8 showed that compared with the negative control group, the supernatant released from the control group had no effect on the MG-63 proliferation at different time points, while the supernatant released from the experimental group could inhibit the MG-63 proliferation at different time points (P<0.05), and the inhibitory effect became more and more obvious with the prolongation of release time. (3) RT-PCR findings showed that compared with the control group, the supernatant released from the experimental group could increase Bax mRNA expression and decrease Bcl-2 mRNA expression at the same time point. To conclude, the PLA electrospun fiber membranes carrying gemcitabine hydrochloride can sustainably inhibit MG-63 proliferation and promote cell apoptosis.

OBJECTIVETo study expression of mutant p53 protein in workers occupationally exposed to benzidine and bladder cancer patients.

METHODSMutant p53 protein in serum from the workers occupationally exposed to benzidine and bladder cancer patients were determined with Immuno-PCR, while exfoliated urothelial cells in the urine samples were classified with Papanicolau grading.

RESULTSPositive rate of mutant p53 protein increased with the exposed intensity index in workers occupationally exposed to benzidine. The positive rate of mutant p53 protein in bladder cancer patients (83.3%) was significantly higher than that in the group 1 of exposed intensity index. The average scanning integrals of PCR amplified band in the group of bladder cancer patients and group 2 of exposed intensity index were both higher than that in the group 1 significantly. Workers in the groups of different exposed intensity indices were further stratified according to Papanicolau grades. In the group 2 of exposed intensity index, the average scanning integrals of PCR amplified band in the stratum of Papanicolau grade II and III were significantly higher than that in the strata of Papanicolau grade I. And in the group 3 of exposed intensity index, the positive rate of mutant p53 protein in the strata of Papanicolau grade III was higher than that in the strata of Papanicolau grade I significantly.

CONCLUSIONThe increase of exposed intensity may not only result in the positive rate of mutant p53 protein, but also the quantity of mutant p53 protein in serum within the low range of benzidine exposure. Once the exposed intensity was beyond that spectrum, the positive rate of mutant p53 protein in serum and the average scanning integrals of PCR amplified band were no longer enhanced with the increase of exposed intensity. There was tight correlation between Papanicolau grade of exfoliated urothelial cells and the positive rate or the quantity of mutant p53 protein for the higher benzidine exposure intensity.

Adult ; Aged ; Aged, 80 and over ; Benzidines ; toxicity ; Female ; Humans ; Male ; Middle Aged ; Occupational Exposure ; Tumor Suppressor Protein p53 ; metabolism ; Urinary Bladder Neoplasms ; metabolism ; Urothelium ; metabolism

OBJECTIVETo detect sperm plasma membrane integrity (PMI) of cigarette smoking infertile males using SYBR-14/ PI fluorescent staining and flow cytometry and investigate its clinical significance.

METHODSWe collected semen samples from 132 cigarette smoking infertile men and 70 normal fertile controls, the former divided into a heavy-smoker group (> 20 cigarettes a day, n = 68) and a light-smoker group (< or = 20 cigarettes a day, n = 64). We performed computer-assisted semen analysis of the semen samples, and determined sperm PMI by flow cytometry after rinsing with PBS and staining by SYBR-14/PI, the sperm with normal PMI indicated as the percentage of those emitting green fluorescence (SYBR-14+/PI- %), dead sperm as the percentage of those emitting red (SYBR-14-/PI+), and moribund sperm as the percentage of those emitting both green and red (SYBR-14+/PI+).

RESULTSBoth the heavy- and light-smoker groups showed significant differences in SYBR-14-/PI+ % and SYBR-14+/PI- % from the normal controls (P < 0.01 or P < 0.05). SYBR-14+/PI- % was remarkably lower, while SYBR-14-/PI+ % markedly higher in the heavy-smoker than in the light-smoker group (P < 0.05). There was a significant correlation between SYBR-14+/PI- % and sperm motility (r = 0.938, P = 0.000).

CONCLUSIONSYBR-14/PI fluorescent staining and flow cytometry analysis could quickly and exactly detect sperm PMI. Cigarette smoking reduces sperm PMI and consequently sperm motility, which might be an important factor of male infertility.

Adult ; Case-Control Studies ; Cell Membrane ; pathology ; Flow Cytometry ; Humans ; Infertility, Male ; Male ; Semen Analysis ; Smoking ; adverse effects ; Spermatozoa ; cytology ; pathology

OBJECTIVETo evaluate the therapeutic effect of Compound Xuanju Capsule (CXC) on autoimmune prostatitis in rat models.

METHODSSixty healthy male Wistar rats were randomly divided into five groups of equal number: blank control, low-concentration purified prostate protein (low-conc PPP), low-conc PPP + CXC treatment, high-concentration PPP (hi-con PPP), and hi-conc PPP + CXC treatment. Autoimmune prostatitis models were established by intragastric administration of PPP solution at 15 mg/ml (low concentration) and 80 mg/ml, respectively. At 30 days after modeling, the rats in the blank control and low-conc and hi-conc PPP model groups were treated with normal saline, and those in the other two groups with CXC at a daily dose of 0.068 g/ml. At 30, 45, and 60 days, all the animals were sacrificed for observation of pathological changes in the prostate tissue and determination of the levels of IL-8, IL-10, and TNF-alpha in the serum.

RESULTSCompared with the PPP models, the hi-conc PPP + CXC group showed significantly reduced levels of IL-8 and TNF-alpha in the serum at 45 days ([148.54 +/- 17.23] and [62.14 +/- 5.59] pg/ml vs [100.77 +/- 11.08] and [32.63 +/- 2.91] pg/ml, P < 0.05) and at 60 days ([143.69 +/- 17.28] and [59.38 +/- 5.50] pg/mlvs [95.77 +/-10.53] and [29.63 +/- 2.66] pg/ml, P < 0.05), and so did the low-cone PPP + CXC group at 45 days ([128.47 +/- 12.21] and [40.43 +/- 3.64] pg/ml vs [111.76 +/- 10.07] and [35.44 +/- 3.17] pg/ml, P < 0.05) and at 60 days ([131.07 +/- 10.93] and [43.34 +/- 3.91] pg/ml vs [97.46 +/- 8.75] and [30.44 +/- 2.75] pg/ml, P < 0.05). The serum level of IL-10 was remarkably elevated in the hi-cone PPP + CXC group as compared with that of the PPP models at 45 and 60 days ([189.14 +/- 16.78] and [184.14 +/- 15.89] pg/ml vs [230.48 +/- 29.96] and [248.48 +/- 31.03] pg/ml, P < 0.05), and so was it in low-cone PPP + CXC group ([223.14 +/- 17.87] and [224.14 +/- 17.93] pg/ml vs [231.42 +/- 23.18] and [249.42 +/- 24.97] pg/ml, P < 0.05). Pathological examination revealed morphological damages to the prostate tissue and infiltration of inflammatory cells in the model rats, but no obvious changes in the normal controls. At 15 days of treatment, the rats in the PPP + CXC group showed enlarged prostate glandular cavity, mild proliferation of epithelial cells, no obvious infiltration of inflammatory cells in the interstitial tissue, and a few visible fibrous tissues under the light microscope.

CONCLUSIONCompound Xuanju Capsule is efficacious on autoimmune prostatis in rats by reducing inflammatory changes in the prostate tissue and improving the expression of inflammatory factors.

Animals ; Autoimmune Diseases ; blood ; chemically induced ; drug therapy ; Capsules ; Interleukin-10 ; blood ; Interleukin-8 ; blood ; Male ; Prostatic Hyperplasia ; pathology ; Prostatic Secretory Proteins ; Prostatitis ; blood ; chemically induced ; drug therapy ; Random Allocation ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo establish a rat model of autoimmune prostatitis using purified prostatic proteins (PPP).

METHODSThirty-six male Wistar rats were randomized into three groups of equal number to receive intramuscular injection of normal saline (normal control group) and PPP at 15 mg/ml (low-concentration group) and 80 mg/ml (high-concentration group). At 4 weeks after modeling, the rats were sacrificed for HE staining of the prostate tissue and examination of the inflammatory factors IL-8 and IL-10 in the serum, immunoglobulins IgA and IgM, and regulatory T cells Th1/Th2.

RESULTSThree rats died in the high-concentration PPP group but none in the low-concentration PPP and normal control groups. Gross observation of the prostate showed increased volume and hard texture of the prostate in the two PPP groups, but no significant change in the normal controls. Pathological examination exhibited morphological damage to the prostatic tissue and inflammatory cellular infiltration in the experimental rats. The serum level of IL-8 was significantly higher in the low- and high-concentration PPP groups ([129.07 +/- 11.48] and [147.58 +/- 17.70] pg/ml) than in the control ([94.12 +/- 7.04] pg/ml) (P < 0.05), while that of IL-10 was remarkably lower in the former two groups ([227.14 +/- 18.19] and [187.14 +/- 16.32] pg/ml) than in the latter ([252.48 +/- 21.72] pg/ml, P < 0.05). The serum level of IgA was markedly elevated in the low- and high-concentration PPP groups as compared with that in the control ([0.25 +/- 0.37] and [0.31 +/- 0.42] vs [0.19 +/- 0.14] mg/ml, P < 0.05), and so was that of IgM ([0.23 +/- 0.41] and [0.34 +/- 0.58 ] vs [0.17 +/- 0.33] mg/ml, P < 0.05). No significant changes were observed in the levels of regulatory T cells Th1/Th2.

CONCLUSIONBoth low and high concentrations of purified prostatic proteins can be used for the construction of autoimmune prostatitis models in rats, while low concentration is preferable for its advantages of lower mortality of the rats and inducement of more consistent manifestations of autoimmune prostatitis.

Animals ; Autoimmune Diseases ; blood ; chemically induced ; pathology ; Disease Models, Animal ; Humans ; Interleukin-10 ; blood ; Interleukin-8 ; blood ; Male ; Prostatic Secretory Proteins ; administration & dosage ; pharmacology ; Prostatitis ; blood ; chemically induced ; pathology ; Rats ; Rats, Wistar

Reproductive toxicity research takes an important place in traditional Chinese medicine pre-clinical safety evaluation. Modern reproductive toxicity experiment includes drug-related miscarriage, fetal death, teratism, and adverse effects on fertility, genital system, embryonic development and fetus, which is different from contraindicated in pregnancy in traditional Chinese medicine theory. Now the three-phases reproductive toxicity study is the method mainly applied in traditional Chinese medicine reproductive toxicity evaluation. Besides that, alternative methods of whole embryos culture and embryonic stem cell test are also used in traditional Chinese medicine embryo toxicity evaluation. This article reviews research progress and pre-clinical evaluation on reproductive toxicity of traditional Chinese medicine.
Animals ; Drug-Related Side Effects and Adverse Reactions ; Drugs, Chinese Herbal ; toxicity ; Embryonic Development ; drug effects ; Embryonic Stem Cells ; drug effects ; Female ; Humans ; Medicine, Chinese Traditional ; Pregnancy ; Reproduction ; drug effects ; Toxicity Tests