OBJECTIVETo observe the effect of Zhibai Dihuang Decoction (ZDD) on mRNA and protein expressions of transient receptor potential family vanilloid subtype 1 (TRPV1) and transient receptor potential family vanilloid subtype 5 (TRPV5) in Ureaplasma urealyticum (UU)-infected rat semens and spermatogenic cells, and to explore the pathomechanism of UU-infected infertility and the intervention of ZDD.

METHODSTotally 45 were randomly selected from 60 4-5 months old SD rats. UU testicular infected animal models were set up after bladder inoculation of UU suspension. The remaining 15 rats were simultaneously injected with normal saline as a normal control group. After a successful modeling, UU infected model rats were randomly divided into the model group, the azithromycin group, and the ZDD group, 15 in each group. Rats in the ZDD group were administered with ZDD at the daily dose of 1 g/kg by gastrogavage. Rats in the azithromycin group were administered with azithromycin suspension at the daily dose of 0. 105 mg/kg by gastrogavage. Equal volume of normal saline was administered to rats in the normal control group and the model group. All medication was performed once daily for 21 successive days. Testes and epididymis were extracted after rats were killed and UU positive rates were compared among all groups. Sperm cells were separated using a mechanical separation technique. Sperm motility parameters were detected using color sperm motion detection system. mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells were determined by real-time quantitative PCR and Western blot.

RESULTSThe UU positive rate was obviously higher in the model group than in the normal control group [(86.7% (13/15 cases) vs. 0] P < 0.05). It was lower in the ZDD group [33.3% (5/15 cases)] and the azithromycin group [26.7% (4/15 cases)] than in the model group (P < 0.05). Compared with the normal control group, class A and B sperms were reduced, the linear velocity and the average velocity were significantly lowered, mRNA and protein expressions of TRPV1 and TRPV5 in spermated genic cells significantly decreased in the model group (P < 0.05). Compared with the model group, class A and B sperms were increased, linear and curve velocities and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group and the azithromycin group (P < 0.05, P < 0.01). Compared with azithromycin group, class A and B sperms were increased, the linear velocity and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group (P < 0.05, P < 0.01).

CONCLUSIONZDD could fight against UU infection and elevate semen quality, which might be associated with up-regulating mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells.

Animals ; Calcium Channels ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Infertility ; Male ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Semen Analysis ; Sperm Motility ; Spermatozoa ; TRPV Cation Channels ; metabolism ; Testis ; Ureaplasma Infections ; Ureaplasma urealyticum

Objective:To evaluate the endothelial functions and autoregulation capacity of cerebral blood flow in patients with Fabry disease .Methods:Brachial artery vasodilation was assessed in 8 pa-tients with Fabry disease and 14 healthy controls by means of flow-mediated dilation ( FMD) and Nitro-glycerin-mediated dilation ( NMD) .Cerebrovascular reactivity was calculated in terms of breath-holding index ( BHI) and vascular motor reactivity ( VMR) by TCD-CO2 test in 4 patients and 14 healthy con-trols.Results:Compared with the controls , brachial artery vasodilation experiment showed no difference (the patients:FMD 15.94%±5.03% and NMD 23.92%±7.23%, the controls: FMD 14.57%± 5 .84% and NMD 22 .64%±6 .96%) , there was no relationship between FMD or NMD and the age , course of disease , MSSI or enzyme activity .In respect of cerebrovascular autoregulation capacity , there was no difference in anterior circulation , while cerebrovascular reactivity tended to be impaired in posteri-or circulation .Conclusion:Endothelial function showed no decline in patients with Fabry disease , but cerebrovascular autoregulation capacity tended to be impaired in posterior circulation .

ObjectiveTo explore the effects of the conjugate prepared from the cholera toxin B subunit(CB) and nerve growth factor(NGF) on the spatial learning and memory abilities and cholinergic function.MethodsThe conjugate of CB-NGF was prepared by the improved sodium metaperiodate method and nasally administrated to the β-amyloid protein(Aβ25-35) induced amnesic mice for 7 days with 2 dosage (7-5 μg/d、15 μg/d). Spatial learning and memory abilities were evaluated by Morris water maze and cholinergic function was assessed with the choline acetyl transferase (ChAT) immunohistochemical methods.ResultsMorris water maze test showed that the escape latency in Aβ25-35-treated mice prolonged and the staying time reduced in the crossed first quadrant where the platform had been located, compared with the control mice (P<0-01). In addition, the number of ChAT positive neuron declined in the model mice(P<0-001). CB-NGF nasal administration significantly shortened the escape latency and elevated the staying time and number of ChAT positive neuron(P<0-01).ConclusionCB-NGF treatment can improve the spatial and memory performance which may involve the neuroprotection to cholinergic system.

To investigate the effects of sorafenib and octreotide combination treatment on cellular proliferation and explore the underlying molecular mechanisms by using an in vitro cell culture system with the human hepatoma cell line, HepG2. HepG2 cells were treated with different concentrations of sorafenib and octreotide alone or in combination. Untreated HepG2 cells were used as controls. Treatment-induced cytotoxicity was determined with the cell counting kit-8 by Sigma-Aldrich, and rate of apoptosis was detected by flow cytometry. Fluorescent microscopy was used to observe rates of cell growth under the various treatments. Treatment-induced changes in protein expressions were detected by enzyme-linked immunosorbent assay (for vascular endothelial growth factor (VEGF)) and Western blotting (for the Mcl-1 apoptosis mediator and the ERK1/2 and PERK1/2 kinases). Sorafenib and octreotide, used alone or in combination, inhibited proliferation and induced apoptosis in HepG2 cells. Combination treatment was more effective than either mono-treatment (F = 200.398, P less than 0.05). Fluorescent microscopy showed that combination treatment stimulated phosphatidylserine, the marker of early apoptosis, better than either mono-treatment. VEGF expression in cultures exposed to combination treatment was also significantly lower than in mono-treatment or untreated control cultures (F = 1019.725, P less than 0.05). Western blotting showed that octreotide mono-treatment had no effect on Mcl-1 expression (vs. control group; P more than 0.05) and that combination treatment significantly lowered Mcl-1 expression (vs. mono-treatment and control groups; P less than 0.05). None of the treatments affected ERK1/2 expression (all, P more than 0.05), while all treatments significantly lowered PERK1/2 expression (vs. control group; F = 2.401, P less than 0.05) and the combination treatment lowered PERK1/2 significantly more than either mono-treatment (P less than 0.05). Sorafenib and octreotide can inhibit proliferation and induce apoptosis in the human hepatoma cell line, HepG2. Combination treatment is significantly more efficacious (P less than 0.05) and produced synergistic effects. The mechanism underlying this phenomenon may depend on synergistic inhibition of VEGF, the anti-apoptotic protein Mcl-1, and the proliferation-inducing PERK1/2.
Apoptosis ; drug effects ; Benzenesulfonates ; pharmacology ; Cell Proliferation ; drug effects ; Hep G2 Cells ; drug effects ; Humans ; Niacinamide ; analogs & derivatives ; Octreotide ; pharmacology ; Phenylurea Compounds ; Pyridines ; pharmacology

Objective To investigate the value of the Wells score and D-dimer assay in assisting pulmonary perfusion imaging (PPI) for the diagnosis of acute pulmonary embolism (APE). Methods One hundred twenty-one patients with suspected APE were studied from January, 2006 to December, 2008. All patients underwent the Wells score, the quantitative D-dimer assay, chest X-ray photography, and PPI. The diagnostic sensitivity, specificity, positive predictive value and negative predictive value of PPI with the assistance of Wells score and D-dimer assay were calculated. Results Fifty (41.3%) patients were diagnosed with APE. PPI combined with chest X-ray photography (Q/X scan) showed positive results in 49 patients. The sensitivity, specificity, positive predictive value and negative predictive value of the Q/X scan were 86.0% (43/50), 91.5% (65/71), 87.8% (43/49) and 90.3% (65/72), respectively. With assistance of Wells score >4 and D-dimer≥0. 5 mg/L, Q/X scan had a positive predictive value of 100.0% (29/29), for patients with Wells score ≤4 and D-dimer<0.5 mg/L, the negative predictive value for Q/X scan was 100.0% (41/41). Conclusion Combined with Wells score and D-dimer assay, PPI can make accurate diagnosis of APE.

ObjectiveTo investigate the Jiuqiang Naoliqing's (JNQ) histological influence on hearts, brains and kidneys of spontaneous hypertension rats (SHR). MethodsThe rats were randomly divided into four groups: Wistar control group, SHR group, higher dose JNQ treated SHR group(0.530 g/kg) and lower dose JNQ treated SHR group(0.265 g/kg). The treatment lasted five weeks, and the rats' blood pressure were monitored through tail pulse. After the perfusion procedure, rats' hearts, brains and kidneys were rapidly removed in low temperature condition and stored in 10% formalin solution of 4 ℃.Then routine sections were obtained and the slides were stained with HE.ResultsBefore treatment, the blood pressure of SHR groups were distinctly higher than that of the control group(P<0.01), nevertheless, no obvious blood pressure downgrade were observed after three week and five week treatment. Histopathologic study showed: in SHR group,heart with hypertrophic cardiac muscle, proliferative arterial wall and strictured lumina; Cortex with angiostenosis, proliferative vascular wall and large perivascular space; Glomerulus atrophy with hyaline degeneration. These pathologic changes got respective alleviation after five week treatment of JNQ, particularly in higher dose group.ConclusionSHR who got five week treatment of JNQ didn't gain obvious blood pressure downgrade. But the treatment did good to their vital organs and can obviously alleviate hearts, brains and kidneys' pathologic changes.

OBJECTIVETo analyze the therapeutic effect of human neural precursor cells transplantation in treatment of neonates with severe brain injury.

METHODThe transplantation was performed on 6 newborns, one of them was diagnosed as extremely severe carbon monoxide poisoning at 5(th) day after birth; one of them was diagnosed as severe hypoglycemia; the others had asphyxia at birth with Apgar scores from 1 to 3 and were diagnosed as severe neonatal asphyxia, severe hypoxic ischemic encephalopathy according to images, electroencephalogram, biochemical examination and clinical manifestation. With the approval of hospital ethics committee and informed consent of the family members, the newborns received human neural precursor cells transplantation at the 4(th) to 20(th) day after birth. With the agreement of a pregnant woman, forebrain cells were obtained from the forebrain of her 12-week old fetus after spontaneous abortion. The cells from the fetal brain were amplified into human neural precursor cells in vitro and were injected into the cerebral ventricle of the patients.

RESULTOn the 2(nd) day after transplantation, sucking and swallowing reflexes gradually appeared in all the patients, muscular tension was also improved, and convulsion stopped. NBNA scoring in 3 of the patients reached normal level on the 28(th) day after birth. The 6 patients were followed up for 12 months. Four patients were normal in psychomotor development and scores of each scale reached normal level. Two patients have cerebral palsy.

CONCLUSIONhNPCs transplantation is safe and effective in treatment of severe neonatal brain injury. More clinical trials and further observation are needed.

Brain Injuries ; surgery ; Female ; Humans ; Hypoxia-Ischemia, Brain ; surgery ; Infant, Newborn ; Male ; Neural Stem Cells ; cytology ; transplantation

OBJECTIVESTo investigate the therapeutic effect of rehabilitation, arthroscopy and "hybrid technique" for posttraumatic knee stiffness (PTKS), and to make the best choice for the treatment.

METHODSFrom February 2004 to November 2009, 66 patients suffered from PTKS were treated, and the clinical data were studied retrospectively, 36 male and 30 female patients with an average age of 41 years were analyzed, knee stiffness time averaged 15 months (0.5 - 108.0 months), 21 cases of patients were treated with rehabilitation (rehabilitation group), 22 cases of patients with arthroscopy + rehabilitation (arthroscopy group) and 23 cases of patients with mini-invasive "hybrid technique" + rehabilitation (hybrid technique group). For each case, the difference of range of motion (ROM) and hospital for special surgery (HSS) score of the knee before and after the treatment were analyzed statistically. The characters of PTKS including the course of the disease, the degree of extensor mechanism involving, physical examination and other ancillary data were also analyzed. The management methods for PTKS were summarized.

RESULTSTotal 66 cases were followed up ranging from 24.0-72.5 months and the mean time was 34.2 months. The average ROM was improved obviously: rehabilitation group increased from 45° ± 22° to 95° ± 24° (t = -11.2, P < 0.05), arthroscopy group from 47° ± 26° to 118° ± 11° (t = -11.0, P < 0.05) and hybrid technique group from 36° ± 22° to 110° ± 14° (t = -13.4, P < 0.05). Both ROM and HSS score of the knee before and after the treatment for each group showed significant difference statistically (t = -9.1, -6.0, -5.2, P < 0.05). Wound necrosis, tearing, re-fracture and extension lag were not found. According to Judet standard at final follow-up, 15 cases were excellent, 3 cases good and 3 cases normal in rehabilitation group; 15 cases were excellent, 5 cases good and 2 cases normal in arthroscopy group; 14 cases were excellent, 8 cases good and 1 case bad.

CONCLUSIONSPathology of PTKS is complex, satisfactory result could be obtained through individualized treatment program, which were established depend on the course of the disease, the degree of extensor mechanism involving, physical examination and ancillary data. The timely and effective surgical interference followed by a comprehensive rehabilitation program is the key point for satisfied outcome.

Adolescent ; Adult ; Aged ; Ankylosis ; etiology ; surgery ; Arthroscopy ; Female ; Follow-Up Studies ; Humans ; Knee Injuries ; complications ; Knee Joint ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Range of Motion, Articular ; Retrospective Studies ; Treatment Outcome ; Young Adult

BACKGROUND/AIMS: Kruppel-like factor 4 (KLF4) is an epithelial-specific transcription factor primarily expressed in the gastrointestinal tract that mediates growth arrest in the colonic epithelium. We tried to find whether KLF4 expression is associated with the progression and differentiation of colorectal cancer. METHODS: We detected KLF4 expression in 109 colorectal specimens (40 normal appearing mucosa, 7 adenomas, and 62 carcinomas) by immunohistochemistry using a tissue microarray. Western blot and RT-PCR analyses were also performed. RESULTS: The upregulation of KLF4 expression in carcinoma tissue was statistically significant (p<0.05) when compared to normal appearing mucosa. The negative and weak positive staining rates in normal appearing mucosa, adenoma, and carcinoma were 42.5%, 71.4%, and 82.3%, respectively, indicating a decreased degree of KLF4 expression over the course of progressive transformation of normal cells into malignant derivatives. KLF4 protein levels showed no correlation with sex, age, or metastatic state (p>0.05), while KLF4 protein expression correlated with the diagnostic stage (p<0.05). Furthermore, strong KLF4 staining was detected in 22.9% (11/48) and 0% (0/14) of well/moderately and poorly differentiated colorectal cancers, respectively. Our results clearly indicate that KLF4 protein expression significantly correlates with the degree of differentiation in colorectal cancers (p<0.05). KLF4 expression in RKO cells is also upregulated by butyrate, an inducer of differentiation. CONCLUSIONS: Downregulation of KLF4 expression may lead to more poorly differentiated tumors.
Adenoma ; Blotting, Western ; Butyrates ; Colon ; Colorectal Neoplasms ; Down-Regulation ; Epithelium ; Gastrointestinal Tract ; Immunohistochemistry ; Kruppel-Like Transcription Factors ; Mucous Membrane ; Transcription Factors ; Up-Regulation

OBJECTIVETo investigate whether inflammatory stress exacerbates hepatic cholesterol accumulation and liver fibrosis using a C57BL/6J mouse model of chronic inflammation.

METHODSTwelve male C57BL/6J mice were given a high-fat diet (15.0% fat, 1.25% cholesterol, 0.5% cholic acid) and randomly assigned to the normal control group (n=6; subcutaneously injected with 0.5 mL of isotonic saline, every other day for 14 weeks) or the chronic inflammation model group (n=6; subcutaneously injected with of 0.5 mL of 10% casein, every other day for 14 weeks). At the end of week 14, the animals were sacrificed and blood was collected from the left ventricle for serological analysis of inflammatory markers and lipid profile, including serum amyloid A (SAA), interleukin-6 (IL-6), total cholesterol (TC) and free cholesterol (FC), low-density lipoprotein (LDL), and high-density lipoprotein (HDL)). Extracted liver tissues were divided for use in histological analysis (lipid accumulation and fibrosis evaluated by Oil Red O, Sirius red and Masson's trichrome staining) and quantitative fluorescence real-time PCR (to measure b-actin normalized expression of TNF-a MCP1, SREBP-2, LDLr, HMGCoA-r, ATF-6, GRP78, BMP-7, TGF-b, and collagens type I and type IV). Comparisons between groups were made by the two-samples t-test or Satterthwaite t-approximation test, collagen type I and type IV.

RESULTSCompared to the normal control group, the inflammation model group showed elevated serum IL-6 (12.55+/-4.75 vs. 32.41+/-7.42 pg/mL, P less than 0.01), reduced serum TC (14.82+/-1.56 vs. 10.62+/-0.48 mmol/L, P less than 0.01), up-regulated hepatic TNF-a mRNA expression (1.05+/-0.35 vs. 2.12+/-0.72, P less than 0.01), and elevated hepatic TC (12.10+/-2.57 vs. 23.21+/-8.75 mmol/L, P less than 0.05). In addition, the inflammation group showed abnormal lipid deposition, and increased and thickened reticular fibers. The livers of the inflammation group also showed up-regulated mRNA expression of SREBP-2 (normal control: 1.01+/-0.19 vs. 2.63+/-0.13, P less than 0.05), GRP78 (1.07+/-0.47 vs. 2.21+/-0.99, P less than 0.05), TGF-b (1.01+/-0.14 vs. 1.38+/-0.28, P less than 0.05), and collagen type I (1.02+/-0.27 vs. 1.71+/-0.51, P less than 0.05) and down-regulation of BMP-7 (1.01+/-0.15 vs. 0.55+/-0.25, P less than 0.01).

CONCLUSIONActivation of the inflammatory system exacerbates hepatic cholesterol accumulation and hepatic fibrosis in C57BL/6J mice.

Animals ; Cholesterol ; metabolism ; Disease Models, Animal ; Fatty Liver ; metabolism ; pathology ; Inflammation ; Liver ; metabolism ; pathology ; Liver Cirrhosis ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred C57BL