OBJECTIVETo apply the Wright-Giemsa stain in micronucleus test and to explore the stain outcomes of Wright-Giemsa dye of various proportions and staining times.

METHODSUse Wright-Giemsa dye, Wright dye (staining time 3 min) and Giemsa dye (staining time 5 min) to stain HepG2 and then observe the staining effect. The Wright-Giemsa dye was applied under 5 different proportions (3:1-1:3) and different staining times (1, 3, 5, 10, 15 min).

RESULTSAfter stained for 3-5 min with the proportion ratio of 3:1 of Wright-Giemsa dye, the HepG2 cells showed much better staining outcomes compared with the single stain of either Wright or Giemsa.

CONCLUSIONSWright-Giemsa stain can be used in cell micronucleus test to obtain good staining outcomes.

Azure Stains ; Coloring Agents ; Hep G2 Cells ; Humans ; Micronucleus Tests ; Staining and Labeling ; methods

OBJECTIVETo investigate the effect of Xianxiong decoction on the mice with acute lung injury induced by lipopolysaccharide.

METHODEighty female ICR mice were randomly divided into 8 groups: model group, Xianxiong decoction group, Daxianxiong decoction group, Xianxiong decoction group without Kansui Radix group, Xianxiong decoction group without Glycyrrhizae Radix et Rhizoma group, Glycyrrhizae Radix et Rhizoma and Kansui Radix group, normal group and control group. Animals of each group, except normal group, were undertaken intraperitoneal injection and intranasal inhalation of lipopolysaccharide (LPS) on day 1, 2, 3 to establish acute lung injury (ALI) model. 30 min after modeling, 0.2 mL corresponding drugs were administrated to each mice, dexam ethasone and normal saline were given to the mice of control group and normal group respectively. White blood cell in blood, neutrophil percentage of blood and bronchoalveolar lavage fluid (BALF) supernatant, the ratio of wet and dry lung tissue ( W/D), histopathological changes of lung tissue were estimated. Sixty ICR mice were randomly divided into normal, model, control, high, middle and low dose Xianxiong decoction groups and were modeled in the same way. ELISA was applied to detect the level of NF-kappaB, TNF-alpha and IL-6 in BALF, PCR for NF-kappaB and TNF-alpha mRNA in lung tissue, and Western blot for NF-kappaB and TNF-alpha. Half of 20 ICR mice were administrated with Xianxiong decoction of its maximum tolerant normal saline.

RESULTCompared with model group, the number of WBC in blood of Xianxiong decoction group mice decreased (P < 0.01), percentage of neutrophils in both blood and BALF decreased as well (P < 0.01, P < 0.05); it also significantly reduced the ratio of W/D (P < 0.01); and found the alveolar wall, the number of inflammatory cells infiltrating improved, compared with model group. Xianxiong decoction reduced the level of NF-kappaB, TNF-alpha and IL-6 in BALF (P < 0.01, P < 0.01, P < 0.05); its high and low dose groups only found TNF-alpha level declined. Five mice died 24 h after administration of Xianxiong decoction which indicated its toxicity when other influential factors were considered.

CONCLUSIONXianxiong decoction is effective on the ALI mice induced by LPS, but it is of toxicity at 3 g x mL(-1).

Acute Lung Injury ; drug therapy ; genetics ; metabolism ; pathology ; Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Interleukin-6 ; genetics ; metabolism ; Lipopolysaccharides ; adverse effects ; Lung ; drug effects ; metabolism ; pathology ; Mice ; Mice, Inbred ICR ; NF-kappa B ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

0.05).However,the shorter the history of the disease,the better the efficacy of the treatment.The younger the patient was,the better the efficacy of the treatment.Conclusions Vulva dystrophy can be treated with focused ultrasound effectively and safely.This approach appears to be a new promising treatment method.

OBJECTIVETo establish a rat model mimicking erectile dysfunction following radical prostatectomy by crush injury or reaction of the cavernous nerve (CN).

METHODSThirty rats were randomized into CN crush group, CN resection group and sham-operated group. Four weeks after surgery, the rat models were evaluated by apomorphine test and ICP/MAP measurement. Fluorogold (FG) retrograde tracer was used to assess the CN injury. The penile tissues were then harvested for immunohistochemical detection of the nNOS-positive fibers to evaluate the CN injury.

RESULTSThe rats in CN crush group and CN resection group exhibited erectile dysfunction in apomorphine test or in response to electrical stimulation of the ganglion stellatum (MPG). In the sham-operated group, the rats showed normal erectile function with increased ICP/MAP following electrical stimulation (P<0.05). Immunohistochemistry revealed reduced nNOS-positive fibers in both CN crush group and CN resection group as compared with those in the sham-operated group (P<0.05), showing no significant difference between the former two groups (P>0.05). The FG-positive MPG cells in CN crush group and CN resection group were significantly less than that in the sham-operated group (P<0.05), and the positive cells were even less in CN resection group (P<0.05).

CONCLUSIONThe rat CN is structurally similar to human CN, and crush injury and resection of the CN are both reliable methods for establishing rat models of erectile dysfunction following radical prostatectomy.

Animals ; Disease Models, Animal ; Erectile Dysfunction ; etiology ; Male ; Nerve Crush ; Penis ; innervation ; Postoperative Period ; Prostatectomy ; adverse effects ; Rats ; Rats, Sprague-Dawley

BACKGROUNDMany researchers studied the possibility of using stem cells as gene therapeutic vector. But few related reports on the adipose tissue-derived stem cells (ADSCs) are available. Therefore we intended to construct a lentiviral VEGF(165) expression vector and then infect the ADSCs to produce therapeutic seed cells.

METHODSEHS1001-68950485313912 clone was mutated by PCR method to produce consensus fragment of VEGF(165) transcript (NM_001025368). Lentivirus was enveloped with pGC-FU, pHelper 1.0 and pHelper 2.0 plasmids in 293T cells. And then the ADSCs (multiplicity of infection = 20) were transfected with the vectors after titer determination. Stable expression of VEGF(165) in ADSCs was confirmed by immunofluorescence staining, enzyme-linked immunosorbent assay (ELISA) and Western blotting analysis.

RESULTSDNA sequencing and 293T transfection verified VEGF(165) was linked to the GFP fused vector. The virus titer is up to 2 × 10(8) determined by quantitative PCR. VEGF(165) transduced cells could show green fluorescence confirmed by immunofluorescence staining (almost 95%). ELISA analyses could detect out the density of VEGF was 850.86 - 1202.13 pg/ml (mean (923.00 ± 31.22) pg/ml) in the supernatant of VEGF(165)-transduced cells but not detected in the GFP-transduced cells (P < 0.001) and the Western blotting analyses also confirmed VEGF(165) expression in VEGF(165)-transduced cells.

CONCLUSIONSThe VEGF(165) over-expression ADSCs were obtained and may be used as a cell therapeutic tool and may be applied for vascular regeneration, especially in the treatment of erectile dysfunction.

Adipose Tissue ; cytology ; Animals ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; Lentivirus ; genetics ; Rats ; Stem Cells ; chemistry ; Vascular Endothelial Growth Factor A ; analysis

OBJECTIVESTo explore the feasibility of DC being in vitro induced from AML cells with cytokine cocktails and their biological properties.

METHODSAML cells were cultured in either presence or absence of cytokine cocktails. DC were studied for morphology, and cytochemical and immunofluorescent staining. Functions of DC were examined by MLC, FITC-conjugated dextran uptake test, and LDH release assay. RT-PCR and FISH were used to analyze the specific fusion genes of culture-derived DC.

RESULTSClassical DC morphological changes occurred in all 15 cultured AML cells. DC-associated surface molecules such as CD(1a), CD(80), CD(86), CD(106), CD(83) and HLA-DR were upregulated (P < 0.05). The allostimulatory abilities of culture-derived DC were significantly higher than those of AML cells uncultured or cultured in the absence of cytokines (P < 0.05). Culture-derived DC only in the presence of GM-CSF + IL-4 have phagocytotic activities. CTL assay was performed in 5 of the 15 samples. At effector/target ratio of 20:1, auto-T lymphocytes primed with the culture-derived DC exhibited no more killing activity to auto-AML cells than those stimulated by IL-2 or uncultured AML cells. Culture-derived DC presenced the native AML-specific aberrant karyotype and related fusion gene.

CONCLUSIONSCytokine cocktails could in vitro induce AML cells into DC with classical morphology, immunophenotype and function. DC maturity induced by different cytokine cocktails could be variable. Culture-derived DC were originated from the native AML cells. AML cells could make the auto-T lymphocyte anergy.

Adolescent ; Adult ; Aged ; Cell Differentiation ; drug effects ; Cytokines ; pharmacology ; Dendritic Cells ; cytology ; Drug Synergism ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; In Vitro Techniques ; Interleukin-4 ; pharmacology ; Leukemia, Myeloid, Acute ; immunology ; pathology ; Male ; Middle Aged ; Monocytes ; cytology ; drug effects ; Tumor Necrosis Factor-alpha ; pharmacology

This paper presents a novel monitor which uses ARM controller AT91SAM7S64 as its main processor, LCM (Liquid Crystal Display Module) for displaying ECG waves, SD (Secure Digital memory) card for data storage and RF module PTR8000 for radio data transmission. This portable monitor boasts alarm function for abnormality and can provide dynamic ECG monitoring for patients.
Computer Communication Networks ; Electrocardiography ; instrumentation ; Humans ; Monitoring, Physiologic ; instrumentation ; Signal Processing, Computer-Assisted ; instrumentation ; Telemetry ; instrumentation ; methods

Background Many researchers studied the possibility of using stem cells as gene therapeutic vector. But few related reports on the adipose tissue-derived stem cells (ADSCs) are available. Therefore we intended to construct a lentiviral VEGF165 expression vector and then infect the ADSCs to produce therapeutic seed cells.Methods EHS1001-68950485313912 clone was mutated by PCR method to produce consensus fragment of VEGF165 transcript (NM_001025368). Lentivirus was enveloped with pGC-FU, pHelper 1.0 and pHelper 2.0 plasmids in 293T cells.And then the ADSCs (multiplicity of infection=20) were transfected with the vectors after titer determination. Stable expression of VEGF165 in ADSCs was confirmed by immunofluorescence staining, enzyme-linked immunosorbent assay (ELISA) and Western blotting analysis.Results DNA sequencing and 293T transfection verified VEGF165 was linked to the GFP fused vector. The virus titer is up to 2x10a determined by quantitative PCR. VEGF165 transduced cells could show green fluorescence confirmed by immunofluorescence staining (almost 95％). ELISA analyses could detect out the density of VEGF was 850.86-1202.13pg/ml (mean (923.00±31.22) pg/ml) in the supernatant of VEGF16s-transduced cells but not detected in the GFP-transduced cells (P ＜0.001) and the Western blotting analyses also confirmed VEGF165 expression in VEGF165-transduced cells.Conclusions The VEGF165 over-expression ADSCs were obtained and may be used as a cell therapeutic tool and may be applied for vascular regeneration, especially in the treatment of erectile dysfunction.

OBJECTIVETo investigate the effect of Compound Xuanju Capsule (CXC) combined with bromocriptine on hyperprolactinemia-induced erectile dysfunction (ED).

METHODSWe randomly assigned 46 patients with hyperprolactinemia-induced ED to receive bromocriptine (trial group, n = 23) and bromocriptine plus CXC (control group, n = 23), respectively, both for 12 weeks. Then we compared the two groups of patients in erectile function and the levels of serum prolactin and testosterone.

RESULTSAfter 12 weeks of treatment, the IIEF-5 scores were significantly improved in both the trial and the control groups as compared with the baseline (19.5 +/- 4.1 vs 13.0 +/- 3.8 and 16.4 +/- 3.7 vs 13.7 +/- 3.5, P<0.05), the level of serum prolactin was remarkably decreased ([156.07 +/- 26.31] vs [478.35 +/- 62.28] mIU/L and [164.73 +/- 28.58] vs [445.26 +/- 57.83] mIU/L, P<0.05), while the level of serum testosterone was markedly increased ([15.34 +/- 5.27] vs [3.80 +/- 1.09] nmol/L and [12.02 +/- 2.36] vs [4.07 +/- 1.25] nmol/L, P<0.05). Post-treatment erectile function was significantly better in the trial than in the control group (P<0.05), and the post-treatment serum testosterone level remarkably higher in the former than in the latter (P<0.05), but there was no significant difference in the serum prolactin level after treatment between the two groups (P>0.05).

CONCLUSIONThe combination of Compound Xuanju Capsule and bromocriptine is highly effective in the treatment of hyperprolactinemia-induced ED, and its effect is even better than that of bromocriptine alone.

Adult ; Bromocriptine ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Erectile Dysfunction ; drug therapy ; etiology ; Humans ; Hyperprolactinemia ; complications ; drug therapy ; Male ; Phytotherapy

OBJECTIVETo explore the possibility of further improvement of the efficacy of neoadjuvant chemoradiotherapy in locally advanced lower rectal cancer and the management of patients with clinical complete regression.

METHODSFrom May 2001 to August 2007, 192 cases with locally advanced lower rectal cancer (T3/T4 or N(+)) received preoperative radiotherapy 40 - 46 Gy/20 - 23 fractions and concomitant oral capecitabine 625 mg/m(2) bid for 10 weeks prior to surgery. Curative resection with total mesorectal excision (TME) was carried out 6 weeks after the end of radiation.

RESULTSAs a result, 117 cases (60.9%) experienced adverse events but only 2 suffered from G3 side effects. Seventeen cases (8.9%) had a clinical complete tumor regression without surgery; 175 patients underwent curative resection, of them 134 cases with low anterior resection (LAR), 32 cases with ultra-low anterior resection with Park's coloanal anastomosis (6 cases with diverting temporary colostomy) and 9 cases with abdominal pelvic resection (APR). Sphincter preservation was achieved in 94.9%. Twenty-four patients (12.5%) got pathological complete response (CR), 17 patients with clinical CR and the overall CR rate was 21.4%. According to the pathological staging post operation: T0N0 41 cases, T2N0 43 cases, T3N0 77 cases, T4N0 5 cases, T2N1 11 cases, T3N1 13 cases, T4N1 2 cases; Graded under Dworak's tumor regression: TRG0 8 patients, TRG1 32 patients, TRG2 28 patients, TRG3 83 patients and TRG4 24 patients, with an overall pathological tumor downstaging in 77.14%. No operative death occurred, 5 patients suffered from rectovaginal fistulas and 4 anastomotic leakages with an overall anastomotic leakage rate of 5.1% (9/175) and all the patients recovered uneventfully after properly managed. All patients were followed up for a median time of 42 months (range, 12 - 87 months). During the time, 11 patients developed lung metastases, 6 liver metastases and 7 had local recurrences. The 3 years disease-free survival (DFS) was 86.6% and overall survival (OS) was 92.6%.

CONCLUSIONSNeoadjuvant chemoradiotherapy has high efficacy in locally advanced lower rectal cancer, resulting in tumor down-staging, improved resectability and sphincter preservation, and reduced local recurrences. Meanwhile the cases with clinical complete response can be followed up closely and safely without surgery.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neoadjuvant Therapy ; Rectal Neoplasms ; drug therapy ; radiotherapy ; Treatment Outcome