This study was purpose to investigate the expression of XIAP mRNA in chronic myeloid leukemia (CML) and to explore its significance in the advance and prognosis of CML. The chromosomal karyotype analysis and detection of XIAP mRNA were performed by the technique of chromosomal R banding and real time PCR in 71 patients with CML and 10 healthy controls. The results showed that there was a significant increase of XIAP mRNA expression in accelerated and blastic phase of the CML, compared with the patients in chronic phase (t = 9.10, 9.30, p < 0.01). Moreover, the difference of XIAP mRNA expression level was not statistically significant in different karyotype groups. It is concluded that the XIAP gene expression in accelerated and blastic phases of CML patients obviously increases, XIAP gene is closely related to the advance of CML.
Adult ; Case-Control Studies ; Female ; Gene Expression ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; pathology ; Male ; Prognosis ; X-Linked Inhibitor of Apoptosis Protein ; genetics

OBJECTIVETo investigate the expression and significance of CD151 in pituitary adenomas.

METHODSThirty-six pituitary adenomas were collected immediately after surgery together with five normal pituitary tissue. Real time-PCR, Western blot and immunohistochemistry analysis were performed to detect the expression of CD151 mRNA and protein in thirty-six pituitary adenomases and five normal pituitary tissues.

RESULTSThe expression of CD151 in all pituitary adenomases was observed to be significantly higher than that in normal pituitary tissues by Western blot, real time PCR, and immunohistochemistry analysis (P < 0.01). The expression levels of protein and mRNA in invasive pituitary adenomas were much higher than those in non-invasive pituitary adenomas (P < 0.01).

CONCLUSIONThe results suggested that the expression of CD151 was closely correlated with malignant degree of pituitary adenomas, which indicated the expression of CD151 was intimately correlated with occurrence and development of pituitary adenomas. Detecting CD151 might be a vital index to predict prognosis of pituitary adenomas.

Adenoma ; metabolism ; Blotting, Western ; Humans ; Immunohistochemistry ; Pituitary Gland ; pathology ; Pituitary Neoplasms ; metabolism ; Prognosis ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Tetraspanin 24 ; metabolism

OBJECTIVETo investigate the feasibility of in vivo magnetic resonance imaging (MRI) tracking of transplanted adipose-derived stem cells (ADSCs) labeled with superparamagnetic iron oxide (SPIO) in rat heart.

METHODSADSCs were labeled with poly-L-lysine (PLL)-SPIO complexes. Intracellular iron uptake was identified by Prussian blue stain and transmission electromicroscopy. Trypan blue staining was used to test the viability of the labeled cells. In vitro MRI of labeled cells was performed. SPIO-labeled ADSCs were transplanted into normal rat hearts and were in vivo imaged with MRI. Image findings on MRI were correlated with histological findings of the rat hearts.

RESULTSThe labeling efficacy of ADSCs with PLL-SPIO was nearly 100%. Light microscopy revealed the SPIO particles were located in the cytoplasm of the ADSCs by Prussian blue staining. Transmission electromicroscopy revealed that the SPIO particles were located in the endosomes in the cytoplasm. There was no significantly deference in viability between labeled and unlabeled groups demonstrated by Trypan blue test (P > 0.05). MRI showed signal loss in gel mixed with labeled cells as compared with the unlabeled cells group and blank group. Signal void on rat hearts were demonstrated on MRI and were well correlated with histological findings where Prussian-blue-stain positive cells presented.

CONCLUSIONMRI can be used to in vivo track the transplanted ADSCs labeled with SPIO into rat hearts and facilitate to understand the conditions of the labeled cells in the transplanted areas.

Adipocytes ; cytology ; Animals ; Cell Differentiation ; Contrast Media ; administration & dosage ; Dextrans ; administration & dosage ; Feasibility Studies ; Image Enhancement ; methods ; Magnetic Resonance Imaging ; methods ; Magnetite Nanoparticles ; administration & dosage ; Male ; Myocardium ; cytology ; pathology ; Rats ; Rats, Wistar ; Stem Cell Transplantation ; methods ; Stem Cells ; cytology

The paper is to report the development of a method of quantitative analysis of multi-components by high performance liquid chromatography (HPLC) for simultaneously determining paeoniflorin sulfonate (PS), paeoniflorin (PF) and albiflorin (AF) in sulfated Paeoniae Radix Alba. Moreover, the cytotoxicity of paeoniflorin sulfonate by MTT-assay and the acute toxicity of mice by administration of paeoniflorin sulfonate were evaluated. Chromatographic separation of paeoniflorin sulfonate, PF and AF were performed on a SHISEIDO CAPCELL PAK C18 column (250 mm x 4.6 mm, 5 microm) for HPLC and a mixture of acetonitrile and 0.02% phosphoric acid solution (15 : 85) as the mobile phase. As detector a spectrophotometer set at 230 nm; column temperature 30 degrees C; flow rate 1.0 mL x min(-1). The toxicity of paeoniflorin sulfonate was evaluated by in vitro cytotoxicity carried out on mouse and human primary hepatocytes, and by acute oral toxicity test carried out on mice. The calibration curve of paeoniflorin sulfonate, PF and AF revealed linearity in the range of 0.041 8 - 1.045 0, 0.023 5 - 0.587 5, and 0.039 8 - 0.995 0 mg x mL(-1), respectively (r > 0.999 8). The average recovery was ranged from 99.11% to 101.71%, RSD < 2%. Paeoniflorin sulfonate does not have any cytotoxicity to cells at all the tested concentrations (< or = 300 micromol x L(-1)) in the in vitro cytotoxicity assay. The maximum tolerance dose of paeoniflorin sulfonate solution and extraction of Paeoniae Radix Alba to mouse is 5 g x kg(-1) and 80 g x kg(-1) respectively. The contents of these three components in the samples were determined with the developed method. It is a rapid, convenient and accurate method to determine multi-components. The content of PF in sulfated Paeoniae Radix Alba is significantly lower, and there is negative correlationship between the content of paeoniflorin sulfonate and PF. The in vitro cytotoxicity assay and in vivo mouse acute toxicity test showed that there is no obvious toxicity of paeoniflorin sulfonate and water-soluble extract of sulfated Paeoniae Radix Alba.
Animals ; Benzoates ; analysis ; isolation & purification ; toxicity ; Bridged-Ring Compounds ; analysis ; isolation & purification ; toxicity ; Chromatography, High Pressure Liquid ; Glucosides ; analysis ; isolation & purification ; toxicity ; Hepatocytes ; drug effects ; Humans ; Mice ; Mice, Inbred ICR ; Monoterpenes ; Paeonia ; chemistry ; drug effects ; Plant Roots ; chemistry ; drug effects ; Plants, Medicinal ; chemistry ; drug effects ; Sulfur ; pharmacology ; Volatilization

OBJECTIVETo compare therapeutic effects of pecking moxibustion therapy and common acupuncture on irritable bowel syndrome (diarrhea type).

METHODSFifty cases of irritable bowel syndrome were randomly divided into a pecking moxibustion group (n=30) and an acupuncture group (n=20). The pecking moxibustion group was treated by pecking moxibustion with a ZHOU's pecking moxibustion pen, and the acupuncture group by common acupuncture. In the two groups, Ganshu (BL 18), Pishu (BL 20), Weishu (BL 21), Shenshu (BL 23), etc. were selected. After treatment of 15 sessions the scores of symptoms and clinical therapeutic effects were compared between the two groups.

RESULTSAfter treatment, the scores of symptoms significantly decreased (both P < 0.01), with a significant difference in the decrease of symptom scores between the two groups (P < 0.05); the total effective rate of 93.3% in the pecking moxibustion group was higher than 75.0% in the acupuncture group (P < 0.05).

CONCLUSIONPecking moxibustion therapy can significantly improve the clinical symptoms of irritable bowel syndrome (diarrhea type), with a better therapeutic effect than common acupuncture.

Acupuncture Points ; Adolescent ; Adult ; Aged ; Diarrhea ; therapy ; Female ; Humans ; Irritable Bowel Syndrome ; therapy ; Male ; Middle Aged ; Moxibustion ; Young Adult

OBJECTIVETo explore the apoptosis and the express of proliferating cell nuclear antigen (PCNA) in spermatogenic cells, and study generation function of the rabbit after scrotal reconstruction with flaps.

METHODSThe 48 New Zealand white rabbits were randomly divided into three groups as experimental group (the scrotum reconstructed with flaps, n = 48), the control group (the sham operated group, n = 24) and the blank group (n = 18). The apoptosis and the expression of PCNA in the spermatogenic cells were detected with TUNEL and the immunohistochemistry from the 3rd to the 8th week after operation. 8 weeks later, 12 animals in each group were fed respectively with one female rabbit to observe the procreation.

RESULTSThe apoptotic index of the spermatogenic cells in blank group was 7.73 +/- 4.95. 3 weeks after operation, the apoptotic index of spermatogenic cells was 22.59 +/- 3.04 in the experimental group, and 21.13 +/- 1.68 in control group, showing no significant difference between the two groups (P > 0.05). 8 weeks after operation, the apoptotic index of spermatogenic cells was 71.85 +/- 2.69 in the experimental group, and 13.64 +/- 2.09 in control group, show a significant difference between them (P < 0.05). The apoptotic index in experimental group increased gradually from the 3rd to 8th week after scrotal reconstruction , which was markedly higher than that in the blank group (P < 0.05). The apoptotic index in control group was higher than that in the blank group at the 3rd week (P < 0.05), but not at the 8th week (P > 0.05). The proliferation index of spermatogenic cells was 9.32 +/- 9.30 and 12.52 +/- 3.87 in experimental group at the 3rd and 4th week, respectively, which was significantly lower than that in blank group (43.07 +/- 2.25) and control group (45.69 +/- 4.98) at the 3rd week (P < 0.05). The proliferation index of spermatogenic cells was 46.98 +/- 18.92 and 49.53 +/- 9.79 in experimental group at the 7th and 8th week, respectively, 39.90 +/- 5.10 in control group at the 8th week, showing no difference between the two groups (P > 0.05). The proliferation index of spermatogenic cells in control group at the 3rd and 8th week was not different from that in the blank group (P > 0.05). The female pairing rabbits in the blank and control group were all pregnant, and the average childbirths were 6.0 +/- 1.28 and 5.92 +/- 1.31 respectively, with no difference between the two groups (P > 0.05). All the female pairing rabbits in the experimental group were not pregnant, showing a significant difference from those in the blank and control group (P < 0.05).

CONCLUSIONSThe rabbit generation functional disturbance after scrotal reconstruction with flaps is due to the excessive apoptosis of spermatogenic cell. The spermatogenic cell proliferation is affected only in the early postoperative period, but can recover later.

Animals ; Apoptosis ; Cell Proliferation ; Male ; Proliferating Cell Nuclear Antigen ; metabolism ; Rabbits ; Scrotum ; surgery ; Seminiferous Epithelium ; cytology ; Skin Transplantation ; Surgical Flaps

OBJECTIVETo study serum gastrin levels in response to early minimal feeding in premature infants and evaluate the clinical effect of early minimal feeding.

METHODSPremature infants with critical score < or = 90 were randomly assigned into two groups: early minimal feeding group (n = 48), non-early minimal feeding group (n = 47). Other premature infants (n = 30) without any complications (critical score > 90) were assigned as normal control group. The premature infants in normal control group were fed with water at 6 h after birth, 1 - 2 ml/kg every time, after once or twice, they were fed with formula, increasing in the amount of formula gradually, until adequate. The premature infants in early minimal feeding group were fed with formula within 72 h after birth, 0.5 - 1 ml/kg, once every 3 h, the amount of formula was increased gradually, until adequate. The premature infants without early minimal feeding were not fed with formula until the illness was stable, the amount of formula was increased gradually until adequate. Situation of gastrointestinal feeding tolerance, growth and development, and clinical symptoms were observed and recorded for the three groups. Serum gastrin levels were monitored at 1, 3, 7 day after birth by radioimmunoassay.

RESULTSSerum gastrin concentrations in the three groups elevated from 1 to 7 days. In early minimal feeding group [(82.4 +/- 24.5) ng/L] and non-early minimal feeding group [(87.0 +/- 40.2) ng/L], the concentrations were significantly higher than those in normal control group [(66.4 +/- 19.7) ng/L] at day 1 (F = 3.36, P < 0.05). At day 3 and 7, the concentrations in early minimal feeding group [(96.3 +/- 14.6) ng/L, (113.0 +/- 16.5) ng/L] were significantly higher than those in non-early minimal feeding group [(73.9 +/- 13.5) ng/L, (92.4 +/- 12.2) ng/L] (P < 0.05). There were significant differences among the three groups in infants with feeding intolerance (2/30, 5/48, 14/47), the period reached full enteral feeding [(20.6 +/- 5.7) d, (27.8 +/- 6.1) d, (39.5 +/- 4.7) d], and in number of hospital day [(29.0 +/- 4.6) d, (39.0 +/- 4.8) d, (48.0 +/- 5.6) d] (P < 0.05). There were significant differences between early minimal feeding group and non-early minimal feeding group in the weight gain three and four weeks after birth [(19.1 +/- 2.4) g/d, (11.9 +/- 3.3) g/d], the period reached birthweight [(19.8 +/- 4.2) d, (25.2 +/- 5.1) d] (P < 0.05). There were no significant difference among the three groups in the weight gain in one and two weeks after birth [(5.9 +/- 2.9) g/d vs. (5.0 +/- 2.1) g/d], the numbers of premature infants with infection, anemia, apnea, or hypoglycemia.

CONCLUSIONEarly minimal feeding in premature infants leads to secretion of gastrin, promotes the development of gastrointestine and may not be associated with occurrence of complications.

Birth Weight ; Enteral Nutrition ; methods ; Female ; Gastrins ; blood ; Gastrointestinal Tract ; growth & development ; Humans ; Infant Nutritional Physiological Phenomena ; Infant, Newborn ; Infant, Premature ; blood ; Infant, Small for Gestational Age ; Male

OBJECTIVETo explore the effect of burying testis in thigh pocket on spermatogenesis.

METHODSGuizhou miniature male pigs at child-bearing period were randomly divided to receive operation of scrotum incision and dissection with the testis burying in thigh pocket (experimental group) or without (control group). 3 months later, testis biopsy was performed on 2 pigs from each group for pathological examination. Then every male pig from both experimental (n = 6) and control group (n = 6) got a mating partner and lived together for 3 months. The fertility of the male pigs was observed. 6 months after operation, testis biopsy was performed again on all the animals from both the groups.

RESULTSBoth at 3 months and 6 months after operation, the pathological examination showed the spermatogenic cells of all stage in contorted seminiferous tubules markedly decreased with no mature sperm in experimental group, while normal spermatogenic cells with mature sperm in control group. After the male pigs lived with mating partners for 3 months, no female pigs staying with the experimental group became pregnant, but the male pigs in control group had a normal fertility.

CONCLUSIONSBurying testis in thigh pocket impedes spermatogenesis in the miniature male pig. So burying testis in thigh pocket is not recommended for patients with scrotum skin defect who wish to remain fertile.

Animals ; Female ; Fertility ; Male ; Pregnancy ; Pregnancy Rate ; Scrotum ; Skin ; injuries ; Spermatogenesis ; Swine ; Swine, Miniature ; Testis ; physiology ; Thigh ; surgery

OBJECTIVETo observe the apoptosis of spermatogenic cells and the expressions of Bcl-2 and Bax proteins after burying the testis in the inguinal pocket, and to investigate their relationship.

METHODSWe randomly divided 36 healthy male New Zealand white rabbits into an experimental group (n = 18) and a control group (n = 18). Models were established by burying testes in the inguinal pocket in the experimental group, while the controls were left untreated. At the end of the 8th week after surgery, 6 animals were randomly taken from each group for measurement of the testis surface temperature and testicular biopsy. The apoptosis of spermatogenic cells in the testis tissues was detected by TUNEL assay, and the expressions of Bcl-2 and Bax proteins determined by immunohistochemistry and imaging analysis.

RESULTSAt 8 weeks after burying the testis in the inguinal pocket, the testicular surface temperature was significantly higher in the experimental group than in the control ([ 38.02 +/- 0.36] degrees C vs [36.15 +/- 0.64 ] degrees C, P < 0.05), and so was the apoptosis index (AI) of spermatogenic cells ([89.69 +/- 3.76] % vs [7.73 +/- 4.95 ] %, P < 0.05). The expression of the Bax protein in the testis was significantly increased, while that of the Bcl-2 protein remarkably decreased in the experimental group as compared with the control group (P < 0.05). The apoptotic cells were mostly primary spermatocytes and round spermatids.

CONCLUSIONElevated local temperature of the testis buried in the inguinal pocket increases the apoptosis of spermatogenic cells, and the spermatogenic cell apoptosis is highly correlated with the decreased expression of Bcl-2 and increased expression of Bax. The changes in the expressions of Bcl-2 and Bax proteins were a main mechanism behind the temperature elevation-induced apoptosis of spermatogenic cells.

Animals ; Apoptosis ; Groin ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rabbits ; Spermatids ; metabolism ; Temperature ; Testis ; metabolism ; pathology ; bcl-2-Associated X Protein ; metabolism

Derris eriocarpa, a traditional Chinese medicine belonging to the family of Leguminosae, is widely distributed mainly over Yunnan, Guangxi and Guizhou of China. Modern pharmacological researches on this herb showed that it had extensive bioactivities, such as promoting urination, removing dampness and cough and reducing inspissated mucus and other biological activities. The extensive studies on the chemical constituents of this plant have resulted in the isolation of triterpenoids, steroids, fatty acid and others, but the flavone compounds haven't reported before. In our further research on the ethyl acetate of this plant, nine flavone compounds were obtained by column chromatography on silica gel, Sephadex LH-20, semi-prep HPLC, polyamide column chromatography and recrystallization for separation and purification. The structures were determined on the basis of extensive spectroscopic analysis, including MS, NMR experiments and comparison with spectroscopic data in the literature, respectively, as diosmetin (1), 3, 3'-di-O-methylquercetin (2), afromosin (3), 6, 3'-dihydroxy-7, 4'-dimethoxyisoflavone (4), odoratin (5), 7, 3'-dihydroxy-8, 4'-dimethoxyisoflavone (6), 6, 4'-dihydroxy-7, 3'-dimethoxyisoflavone (7), 5, 7, 4'-trihydroxy-3, 3', 5'-trimethoxyflavone (8), and alpinumisoflavone (9). All these compounds were isolated from Derris eriocarpa How for the first time. And the in vitro assays showed that compound 2 possessed moderate inhibitory activity against human cancer cells K562 and HEL.
Derris ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; pharmacology ; Humans ; K562 Cells