Objective:To detect the expression of six miRNAs for miR-146a,miR-224,miR-34c,miR-200a,miR-148b,miR-375 level in primary hepatocellular carcinoma patients tissues and serum,and to explore the correlation between the expression of miRNA and inflammatory factors,in the hope of verification whether circulating miRNA could be used as an ideal blood borne biomarker for early detection of primary liver cancer.Methods:Venous blood samples were collected from patients with primary liver cancer,hepatitis,cirrhosis and healthy controls,cancer tissues and para cancerous tissues were collected from patients with cancer.After the total RNA was extracted,the relative expression levels of miRNA were detected by real-time quantitative PCR,and the relationship between the expression level of miRNA and the expression of serum tumor markers AFP,CEA,CA19-9 and CA125 was detected.The correlation between the expression level of miRNAs and the expression of HBsAg,anti-HBs,HBeAg,anti-HBe,anti-HBc and inflammatory factors IL-12,IL-4,IL-6,IL-10,IFN-γ,TNF-α was detected as well.Results:Compared with the healthy group,the expression of miR-34c,miR-224 and miR-146a in serum and tissues of PHC group was significantly up-regulated,and the expression of miR-200a,miR-148b and miR-375 in serum and tissues of PHC group was significantly down regulated.The difference was statistically significant.There was a regression relationship between HBsAg and serum miR-375 and miR-146a,and miR-375 decreased with the increase of HBsAg expression,while miR-146a increased with the increase of HBsAg.There was a regression relationship between IFN-γ and miR-146a,miR-146a increased with the decrease of IFN-γ expression.The diagnostic ability of miR-375 and miR-146a was greater than that of CA19-9 and AFP.Conclusion:MiR-146a,miR-224,miR-34c,miR-200a,miR-148b,miR-375 in serum and tissue of primary hepatocellular carcinoma are differentially express,among them,the diagnostic ability of miR-375 and miR-146a is better than that of AFP and CA19-9,and serum miR-375 and miR-146a may be the new markers for early diagnosis of PHC.

Niemann-Pick disease type C (NPC) is an autosomal recessive lysosomal lipid storage disease associated with impaired intracellular cholesterol trafficking. A wide spectrum of clinical phenotype has been described, with a possible onset at all ages of life from the neonatal period to adulthood, more often in childhood. Typically, hepatosplenomegaly, dystaxia, dysphagia, dysarthria and dementia are presented in NPC patients. Neurologic symptoms vary according to the onset age, but prolonged neonatal cholestasis, splenomegaly, cataplexy and vertical supranuclear gaze palsy are more specific signs to the diagnosis of the disease. Impaired cholesterol trafficking and unesterified cholesterol accumulation in the late endosomes and lysosomals, as a results of mutations in NPC1 or NPC2 genes, are initial for the disease, and defective cellular autophagy, defective lysosomal calcium homeostasis and oxidative stress may all play roles in the physiological processes. The definite diagnosis requires demonstration of unesterified cholesterol accumulated in fibroblasts cultured from skin biopsies or of pathogenic mutation of NPC1/NPC2 genes. Miglustat, the only available treatment approved to date, can alleviate neurological symptoms and slow disease progression when administered earlier.
Diagnosis, Differential ; Humans ; Niemann-Pick Disease, Type C ; diagnosis ; etiology ; genetics ; therapy

This study is to investigate the effect of total flavonoids of Uygur medicine bugloss (BTF) on rats with myocardial ischemia/reperfusion injury, and to explore the mechanisms by which it acts. Left anterior descending (LAD) coronary artery in rats was occluded for 30 min followed by 4 h reperfusion. Meanwhile, BTF dissolved in saline was administered intraperitoneally at dosage of 10, 30 and 50 mg x kg(-1). Electrocardiograph, infarction index, serum myocardial enzymes and heart function were determined to evaluate the effect of BTF. Some other observations were carried out to explore whether inhibiting inflammation and apoptosis is involved in the mechanisms underlying BTF. Our results showed that in ischemia/reperfusion injured rats BTF could dose-dependently reduce myocardial infarction index and myocardial enzyme leakage, and enhance heart function, indicating that it possesses significant cardio protection. ELISA analysis showed that BTF could decrease the content of myocardial inflammatory cytokines such as IL-1beta, IL-6 and TNF-alpha. Western-blotting confirmed that BTF could increase the expression of anti-apoptotic protein Bcl-2 and reduce the expression of proapoptosis protein Bax. Further more, the phosphorylation level of PI3K and Akt was upregulated by BTF treatment. BTF can protect rat against myocardial ischemia/reperfusion injury. Anti-inflammation and inhibition of apoptosis through upregulating PI3K/Akt signal pathway may contribute to the protective effect of BTF.
Animals ; Apoptosis ; Apoptosis Regulatory Proteins ; Boraginaceae ; chemistry ; Flavonoids ; pharmacology ; Heart ; Interleukin-6 ; Myocardial Infarction ; Myocardial Reperfusion Injury ; drug therapy ; Myocardium ; Phosphatidylinositol 3-Kinases ; Phosphorylation ; Protective Agents ; Proto-Oncogene Proteins c-akt ; Rats ; Signal Transduction ; Tumor Necrosis Factor-alpha ; bcl-2-Associated X Protein

OBJECTIVETo verify the clinical efficacy of acupuncture on motor dysfunction in ischemic stroke of subacute stage.

METHODSThe multi-central randomized controlled trial was adopted. One hundred and twenty-six cases of ischemic stroke of subacute stage were randomized into an acupuncture group (61 cases) and a conventional treatment group (65 cases). The basic treatment of western internal medicine and rehabilitation training were applied to the patients of the two groups. In the acupuncture group, acupuncture was supplemented at the body points located on the extensor of the upper limbs and the flexor of the lower limbs. In combination, scalp acupuncture was applied to NS5, MS6 and MS6 on the affected side. The treatment was given 5 times a week and totally 8 weeks were required. The follow-up observation lasted for 3 months. The scores in Fugl-Meyer scale and NIHSS scale and Barthel index were compared between the two groups before treatment, in 4 and 8 weeks of treatment and the 3-month follow-up observation after treatment separately.

RESULTSIn 4 and 8 weeks of treatment and the follow-up observation, Fugl-Meyer scale score was improved obviously in the patients of the two groups (all P<0. 01). In 8 weeks of treatment and the follow-up observation, Fugl-Meyer scale score in the acupuncture groupwas im proved much apparently as compared with that in the conventional treatment group [68. 0 (43. 0,86. 5) vs 52. 5 (30.3, 77.0), 77.0 (49.5, 89.0) vs 63. 0 (33.0, 84.0), both P<0. 05]. Except that NIHSS scale score was not reduced apparently in 4 weeks of treatment in the conventional treatment group (P>0.05), the results of NIHSS scale at the other time points were all decreased obviously as compared with those before treatment in the patients of the two groups (all P<0. 01). In 8 weeks of treatment and the follow-up observation, the results in the acupuncture group were reduced much apparently as compared with those in the conventional treatment group [5. 0 (3.0,8.0) vs 7. 0 (3.0,13.8), 4. 0 (1.5,7.0) vs 6.0 (2.0,11.7) ,both P<0. 05]. In 8 weeks of treatment and the follow-up observation, Barthel index was improved obviously as compared with that before treatment in the patients of the two groups (all P<0. 05). The improvement in the acupuncture group was much more significant as compared with the conventional treatment group [75. 0 (60. 0,87. 5) vs 65. O (36. 3, 87. 5), P<0. 051.

CONCLUSIONBased on the conventional treatment, Acupuncture achieves the satisfactory clinical efficacy on motor dysfunction in ischemic stroke of subacute stage.

Acupuncture Therapy ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Motor Activity ; Stroke ; physiopathology ; therapy ; Treatment Outcome

The aim of this study was to investigate the effect of alanine solution as α-N-acetylgalactosaminidase enzyme reaction buffer on the enzymatic activity of A antigen. The binding ability of α-N-acetylgalactosaminidase with RBC in different reaction buffer such as alanine solution, glycine solution, normal saline (0.9% NaCl), PBS, PCS was detected by Western blot. The results showed that the efficiency of A to O conversion in alanine solution was similar to that in glycine solution, and Western blot confirmed that most of enzymes blinded with RBC in glycine or alanine solution, but few enzymes blinded with RBC in PBS, PCS or normal saline. The evidences indicated that binding of enzyme with RBC was a key element for A to O blood group conversion, while the binding ability of α-N-acetylgalactosaminidase with RBC in alanine or glycine solution was similar. It is concluded that alanine solution can be used as enzyme reaction buffer in A to O blood group conversion. In this buffer, the α-N-acetylgalactosaminidase is closely blinded with RBC and α-N-acetylgalactosaminidase plays efficient enzymatic activity of A antigen.
ABO Blood-Group System ; immunology ; Alanine ; Blood Grouping and Crossmatching ; methods ; Humans ; Solutions ; alpha-N-Acetylgalactosaminidase ; immunology

OBJECTIVETo observe the effect of Yangjing Capsule on the expressions of P-Erk1 and P-Akt1 in the corpus cavernosum of aged rats and to explore its action mechanism.

METHODSForty 24-month-old SD rats were equally randomized into an experimental and a control group, 10 selected from each group for detection of the eNOS expression before medication. The rats in the experimental group (n = 10) were treated intragastrically with Yangjing Capsule, while those in the control with physiological saline, both at 0.5 g per kg body weight per day for 4 weeks. Then all the rats were killed, and the corpus cavernosum tissues harvested for determination of the expressions of eNOS, P-Erk1 and P-Akt1 by HE staining, RT-PCR and immunohistochemistry.

RESULTSObvious distension was observed in the experimental group, but not in the control. The expression of eNOS in the rat corpus cavernosum was significantly higher after treatment than before treatment in the experimental group (24.10 +/- 2.40 vs 18.80 +/- 2.04, P < 0.05), but presented no obvious difference in the control (18.15 +/- 1.98 vs 19.35 +/- 1.50, P > 0.05), and it was remarkably higher in the former than in the latter (P < 0.05). The expression of P-Erk1 was markedly lower in the experimental than in the control rats (0.71 +/- 0.13 vs 0.83 +/- 0.17, P < 0.01), but that of P-Akt1 exhibited no remarkable difference in the same condition of GAPDH between the two groups (0.58 +/- 0.17 vs 0.48 + 0.13, P > 0.01).

CONCLUSIONBoth P-Erk1 and P-Akt were expressed in the corpus cavernosum of the aged rats. Yangjing Capsule could improve the sexual function of aged rats, which might be associated with the lowered expression of P-Erk1.

Aging ; metabolism ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Male ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Penis ; drug effects ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Rats ; Rats, Sprague-Dawley

AIM: To investigate the relationship between cerebroprotection of pinocembrin and epoxyeicosatrienoic acids (EETs) and their regulating enzyme soluble epoxide hydrolase (sEH). METHODS: Rats underwent middle cerebral artery occlusion (MCAO) to mimic permanent focal ischemia, and pinocembrin was administrated via tail vein injection at 10 min, 4 h, 8 h and 23 h after MCAO. After 24 MCAO, rats were re-anesthetized, and the blood and brain were harvested and analyzed. RESULTS: Pinocembrin displayed significant protective effects on MCAO rats indicated by reduced neurological deficits and infarct volume. Importantly, co-administration of 0.2 mg·kg(-1) 14, 15-EEZE, a putative selective EET antagonist, weakened the beneficial effects of pinocembrin. 14, 15-EET levels in the blood and brain of rats after 24 h MCAO were elevated in the presence of pinocembrin. In an assay for hydrolase activity, pinocembrin significantly lowered brain sEH activity of MCAO rats and inhibited recombinant human sEH activity in a concentration-dependent manner (IC50, 2.58 μmol·L(-1)). In addition, Western blot and immunohistochemistry analysis showed that pinocembrin at doses of 10 mg·kg(-1) and 30 mg·kg(-1) significantly down-regulated sEH protein in rat brain, especially the hippocampus CA1 region of MCAO rats. CONCLUSION Inhibiting sEH and then increasing the potency of EETs may be one of the mechanisms through which pinocembrin provides cerebral protection.
Animals ; Arachidonic Acids ; metabolism ; Brain ; drug effects ; enzymology ; metabolism ; Brain Ischemia ; drug therapy ; enzymology ; genetics ; metabolism ; prevention & control ; Disease Models, Animal ; Epoxide Hydrolases ; genetics ; metabolism ; Flavanones ; administration & dosage ; Humans ; Male ; Protective Agents ; administration & dosage ; Rats ; Rats, Sprague-Dawley

To screen potential hamster chymase 2 inhibitors, a high-throughput screening (HTS) model was established. Recombinant hamster chymase 2 with active form was cloned and expressed in E. coli. The HTS model with total volume of 50 microL in 384-well microplate was based on fluorescence analysis and was proved sensitive as well as specific (Z' = 0.84). A total of 40 080 samples (including 28 060 compounds and 12 020 natural products) were screened, and 613 samples with inhibition greater than 90% were selected for further rescreening. Finally, compounds J16647 and J16648 were identified with high inhibitory activity on chymase 2, and whose IC50 values were 0.823 and 0.690 micromol x L(-1), respectively.
Animals ; Chymases ; analysis ; antagonists & inhibitors ; Cricetinae ; Enzyme Inhibitors ; analysis ; pharmacology ; Escherichia coli ; metabolism ; High-Throughput Screening Assays ; methods ; Inhibitory Concentration 50 ; Rats ; Structure-Activity Relationship

OBJECTIVETo search for an effective therapy for functional anejaculation (AE).

METHODSFifty-five AE patients were randomized into a treatment group (n = 30) and a control group (n = 25), the former treated with Yangjing Capsule (once 5 pills, tid) and low-dose tadalafil (5 mg, qd alt, 1 h before bedtime), while the latter with oral ephedrine (25 mg before bedtime). Meanwhile, the patients were advised to expose themselves to sexual stimulation, reduce the frequency of sexual intercourses and quit masturbation. The medication lasted 1-3 months, followed by observation of the therapeutic effects.

RESULTSThe total effectiveness rate was 83.34% in the treatment group, 11 cases cured, 8 obviously improved, 6 improved and 5 unimproved, significantly higher than 40.00% in the control group, 4 cases cured, 3 obviously improved, 3 improved and 15 unimproved (P < 0.05).

CONCLUSIONYangiing Capsule plus low-dose tadalafil is safe and effective for the treatment of functional anejaculation.

Adult ; Capsules ; Carbolines ; administration & dosage ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Infertility, Male ; drug therapy ; Male ; Phytotherapy ; Sexual Dysfunction, Physiological ; drug therapy ; Tadalafil ; Treatment Outcome ; Young Adult

OBJECTIVETo observe the effects of Yixin Kangtai Capsule (YKC) on sperm concentration and motility, superoxide dismutase (SOD) and malondialdehyde (MDA) in aged SD rats, and to explore its possible mechanisms of action.

METHODSWe randomly assigned fifty 24-month-old male SD rats to five groups of equal number: blank control, testosterone undecanoate, high-dose YKC, medium-dose YKC and low-dose YKC, and treated them intragastrically with physiological saline at 10 ml/kg, testosterone un-decanoate suspension at 4 mg/kg, and YKC at 0.6 g/kg, 0.3 g/kg and 0.15 g/kg, respectively, once a day for 30 days. Then we harvested the epididymides, which were stripped of fat and envelope, cut into pieces and placed in the saline at 37 degrees C for 10 to 15 minutes. Then we detected the sperm parameters with the Weili CASA sperm analysis system, measured the levels of serum SOD and MDA, and, following HE staining, observed the histomorphological changes of the testicular tissue under the light microscope.

RESULTSCompared with the controls, the rats treated with high-, medium- and low-dose YKC exhibited significantly increased sperm concentration ([152.3 +/- 24.0] x 10(6)/ml vs [334.5 +/- 97.7], [302.7 +/- 158.9] and [221.4 +/- 75.6] x 10(6)/ml, P < 0.05) and markedly improved sperm motility ([26.5 +/- 2.0]% vs [35.5 +/- 5.9], [33.1 +/- 5.4] and [28.4 +/- 2.1]%, P < 0.01). High-dose YKC significantly increased the level of serum SOD ([22.0 +/- 5.3] U/ml, P < 0.01) and reduced the content of MDA ([13.6 +/- 4.6] nmol/ ml, P < 0.01). Testicular histology showed that YKC induced more active proliferation of cells, and increased the number of dividing cells and that of mature sperm in the lumen, with little interstitium, abundant blood vessels, and well-developed stromal cells.

CONCLUSIONYixin Kangtai Capsule can improve spermatogenesis in aged SD rats by regulating the levels of SOD and MDA.

Aging ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Male ; Malondialdehyde ; blood ; Rats ; Rats, Sprague-Dawley ; Sperm Motility ; drug effects ; Spermatogenesis ; drug effects ; Superoxide Dismutase ; blood