Antiviral Agents ; pharmacology ; Dose-Response Relationship, Drug ; Hep G2 Cells ; Hepatitis B Surface Antigens ; metabolism ; Hepatitis B e Antigens ; metabolism ; Hepatitis B virus ; drug effects ; Humans ; Interferon-alpha ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Recombinant Proteins ; Reverse Transcriptase Polymerase Chain Reaction ; STAT1 Transcription Factor ; genetics ; metabolism

OBJECTIVETo explore the relationship between hypoxia and the apoptosis of corpus cavernosum smooth muscle cells (CCSMC) in SD rats.

METHODSCCSMCs were cultured in vitro and identified by immunohistochemistry, and then underwent hypoxia interference at the concentration of 1% O2 for 12, 24, 48 and 72 hours, with normal oxygen concentration as the control. Flow cytometry was used to determine the cycles and apoptosis of the cells.

RESULTSThe cultured CCSMCs grew well, positive for anti-smooth muscle alpha-actin monoclonal antibody immunohistochemical staining. Flow cytometry showed that the number of CCSMCs in G0/G1 was gradually increased within 48 hours and then decreased, just opposite to the proportion of the S phase cells. But no regular change was found in the proportion of the cells in the G2/M phase.

CONCLUSIONHypoxia promotes the apoptosis of CCSMCs in a time-dependent manner, to the maximum at 48 hours, and then cell lysis may occur, but with no further apoptosis.

Animals ; Apoptosis ; Cell Hypoxia ; Cells, Cultured ; Male ; Muscle, Smooth, Vascular ; cytology ; Penis ; pathology ; Rats ; Rats, Sprague-Dawley

BACKGROUNDRNA interference (RNAi) is a powerful tool to silence gene expression post-transcriptionally. Our previous study has demonstrated that small interfering RNAs (siRNAs) have sufficiently inhibited hepatitis B virus (HBV) replication and expression in vitro. In this study we observed the RNAi-mediated inhibitory effects on HBV replication in mice models and accessed the specificity of these effects.

METHODSA mutant RNAi vector (pSI-C mut) with two base pairs different from the original target gene sequence at the RNAi vector (pSI-C) was constructed according to the method described in this study. A mouse model of acute hepatitis B virus infection was established by injecting naked plasmid pHBV1.3 via the tail vein with acute circulatory overload. pSI-C, pSI-C mut and the irrelevant RNAi control plasmid for green fluorescent protein (GFP) gene, pSIGFP were respectively delivered with pHBV1.3 by tail vein injection method. Six days post injection, enzyme-linked immunosorbent assay (ELISA) assay was used to measure the concentration of HBV surface antigen (HBsAg) in mouse serum, immunohistochemical straining method was used to visualize the expression of HBV core protein (HBcAg) in liver tissues, and the transcriptional level of HBV C mRNA in liver tissues was detected by reverse transcriptase PCR (RT-PCR) analysis.

RESULTSInjection of pSI-C exerted magnificent and specific inhibitory effects on the replication and expression of HBV in the murine model. After 6-day post-injection (p.i.), the OD values were shown to be 5.07 +/- 1.07 in infecting group and 0.62 +/- 0.59 in pSI-C group. The concentration of HBsAg in pSI-C group was significantly lower than that in infecting group (P < 0.01). Liver intracellular synthesis of viral core protein was sharply reduced to 0.9% +/- 0.1%, compared with 5.4% +/- 1.2% of positive hepatocytes in infecting group (P < 0.01), and the transcriptional level of HBV C mRNA was greatly reduced by 84.7%. However, the irrelevant RNAi control plasmid (pSIGFP), and the pSI-C mut did not show the same robust inhibitory effects as pSI-C.

CONCLUSIONpSI-C exert efficient and specific inhibitory effects on HBV replication and expression in mice models.

Animals ; Hepatitis B ; therapy ; virology ; Hepatitis B Core Antigens ; biosynthesis ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; physiology ; Male ; Mice ; Mice, Inbred BALB C ; RNA Interference ; RNA, Messenger ; analysis ; RNA, Small Interfering ; therapeutic use ; RNA, Viral ; analysis ; Virus Replication

BACKGROUND: Eccentric exercise displays many advantages over concentric and isometric contractions, and it has been applied in exercise training and rehabilitation. The existing researches focus on the underlying mechanisms at molecular and neural levels, and the specific adaptation after eccentric exercise may be related to the adaptive signaling pathway. OBJECTIVE: To review the unique physiological characteristics and mechanisms of eccentric exercise, thereby providing reference for in-depth understanding of eccentric exercise. METHODS: The relevant articles were searched in PubMed (1990-2017) and CNKI (2010-2017) with the keywords of "eccentric exercise, negative muscle work, eccentric exercise training, downhill run, concentric exercise, positive muscle work, concentric exercise training, and uphill running" in English and Chinese, respectively. Ultimately, the articles eligible for physiological characteristics, acute response and adaptive mechanism were enrolled. RESULTS AND CONCLUSION: Totally 98 articles were retrieved, and 54 pertinent papers were enrolled for analysis based on the inclusion and exclusion criteria. The mechanical, molecular and neural mechanisms of eccentric contractions differ from those of concentric and isometric contractions. Special metabolic and cardiorespiratory responses after eccentric exercise reflect the advantages in making programs of exercise training and rehabilitation. Particularly, compared with concentric exercise, fewer motor units are recruited during eccentric exercise and exert muscle a greater stimulation, which involves the process of exercise-induced muscle damage and the activation of satellite cells. The content of satellite cells after eccentric exercise is higher than that of concentric exercise, suggesting that satellite cells may play an important role in the muscle damage reconstruction caused by eccentric exercise. In conclusion, eccentric exercise is of great significance for exercise training and rehabilitation, while further study on the adaptive mechanism of eccentric exercise is necessary.

OBJECTIVETo compare the accuracy of preoperative T staging of gastric cancer by oral and intravenous contrast-enhanced gastric ultrasonography.

METHODSOne hundred and forty three patients who had been diagnosed as gastric cancer by endoscopic biopsy and confirmed by pathology after operation were examined by oral and intravenous contrast-enhanced gastric ultrasonography, and they were divided into satisfied group and non-satisfied group according to the 2-D image quality of lesion. The results were compared with postoperative pathologic findings.

RESULTSAll the patients with gastric cancer presented regional gastric wall thickening. Among them, 117 cases were clearly presented with good image quality. The remaining 26 cases were presented with vague profile, the ulcerative surface of lesion was filled with hyperechogenicity combined with rear shadow. The accuracy of oral contrast-enhanced ultrasonography in determining the T stage of gastric cancer was 74.1%. The accuracy in satisfied group and non-satisfied group was 78.6% and 53.8%, respectively. The enhancement pattern of 143 cases was showed as hyperenhancement during the arterial phase and hypoenhancement during the portal phase in DCUS. The accuracy of double contrast-enhanced ultrasongraphy in determining the T stage of gastric cancer was 86.7%, but the accuracy in satisfied group and non-satisfied group was 88.9% and 76.9%, respectively. There was a significant difference between the two methods (χ(2) = 9.031, P < 0.01).

CONCLUSIONDCUS is more accurate than oral contrast-enhanced ultrasonography as a useful diagnostic method for preoperative T staging of gastric cancer.

Adult ; Aged ; Contrast Media ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; methods ; Preoperative Period ; Stomach Neoplasms ; diagnostic imaging ; pathology ; Ultrasonography ; methods

OBJECTIVETo investigate the effects of different subtypes IFN alpha (IFN alpha2b, IFN alpha2a, and IFN alpha1b) transduction molecular STAT1, STAT2, IFNAR, PKR, and RNase L, and to study the differences of their antiviral effects and to evaluate the key signaling transduction molecules.

METHODS(1) After HepG2 cells were treated with IFN alpha2b, IFN alpha2a, or IFN alpha1b, the mRNA levels of STAT1, STAT2, IFNAR, PKR, and RNase L were detected by RT-PCR. (2) After HepG2 cells were treated with 1000 U/ml IFN alpha2b, IFN alpha2a, or IFN alpha1b, the protein expression levels of STAT1 and IFNAR were examined by Western blot.

RESULTSRT-PCR results: (1) IFNAR, STAT1, and STAT2 mRNA expression levels were slightly higher in the IFN alpha1b group than those in the IFN alpha2b group (P > 0.05). The mRNA expression levels in IFN alpha1b or IFN alpha2b groups were significantly higher than in the IFN alpha2a group (P < 0.05). (2) The PKR mRNA expression showed no significant differences among IFN alpha1b, IFN alpha2b, and IFN alpha2a groups. (3) The RNase L mRNA expression was very weak. We could not compare the differences of the RNase L mRNA levels in different groups by RT-PCR. Western blot results: (1) The IFNAR, and STAT1 protein expressions were greatly up-regulated after IFN alpha induction compared with the untreated group (P < 0.05). (2) The IFNAR, and STAT1 protein expression levels in IFN alpha1b group were slightly higher than the IFN alpha2b group. IFNAR, and STAT1 protein levels of IFN alpha1b or IFN alpha2b group were significantly higher than IFN alpha2a group (P < 0.05).

CONCLUSIONSTAT1, STAT2, IFNAR mRNA and protein expressions could all be markedly up-regulated after IFN alpha treatment. Effects of IFN alpha1b or IFN alpha2b were greatly stronger than IFN alpha2a. The PKR mRNA expression also was greatly up-regulated after IFN alpha treatment. Expression levels of PKR in IFN alpha1b, IFN alpha2b, and IFN alpha2a groups were all similar. The mRNA level results were consistent with the protein level results. Our results showed that the antiviral activity of IFN alpha1b or IFN alpha2b were stronger than that of IFN alpha2a. The signal transduction molecules STAT1, STAT2, and IFNAR could be regarded as a key index to evaluate antiviral activity of IFN alpha. Further confirmation is still needed to see whether PKR could be regarded as a key index.

Antiviral Agents ; pharmacology ; Carcinoma, Hepatocellular ; virology ; Humans ; Interferon-alpha ; pharmacology ; Liver Neoplasms ; virology ; Recombinant Proteins ; STAT1 Transcription Factor ; biosynthesis ; genetics ; STAT2 Transcription Factor ; biosynthesis ; genetics ; Signal Transduction ; Tumor Cells, Cultured

OBJECTIVETo study clinical treatment effects of inactivating myofascial trigger points with needling and muscle stretching for the treatment of knee osteoarthritis(OA).

METHODSRetrospective analyses were made to investigate the clinical data of pain clinic outpatient in our hospital from 2010 to 2014, and 108 patients with knee OA, including 35 males and 73 females, were treated with acupuncturing of myofascial trigger points and stretching of muscles and structure around knee. The puncturing of trigger points, and the back and forth movement of needle were required to elicit local twitch response of muscle. After acupuncture treatment, muscle stretch around the knee joint was performed by a therapist. All patients must do homework of self stretching exercise. The extent of stretching should be to gradually increased under a tolerable pain. The ROM and walking pain VAS scores were measured before and after whole therapy and were statistically analyzed during 1 year of follow up.

RESULTSAll the patients were followed up, and 95 patients had no pain after 1 year. The VAS scores were improved from preoperative severe 7.6±0.5, moderate 4.9±0.7, to mild 1.9±0.6 and painless 0.3±0.2.

CONCLUSIONSThe walking pain of knee OA might be alleviated by the acupuncture and stretch to inactivate the myofascial trigger point.

OBJECTIVETo evaluate the quantitative relationship between white matter tract (WMT) variation resulting from cerebral tumors and shifting of the fractional anisotropy (FA) index in magnetic resonance (MR) diffusion tensor imaging (DTI).

METHODSFour female and 8 male patients aged from 21 to 62 years with brain malignancies (2 malignant lymphomas, 2 low-grade astrocytomas, and 8 high-grade cerebral gliomas) underwent conventional contrast-enhanced MR and DTI examinations before operation. Routine T(2)-weighted image, fractional anisotropic (FA) map, color-coded directional map, three-dimensional white matter tractography (WMT), and the FA index of bilateral internal capsule were obtained in every patient.

RESULTSFiber tractography derived from DTI was consistent with known white matter fiber anatomy. The DTI patterns in WMT altered by the tumor were categorized on the basis of FA1/FA2 ratio as follows: pattern 1, FA1/FA2> or =75% with normal or only slightly decreased FA; pattern 2, 50%< or =FA1/FA2<75% with WMT displacement; pattern 3, 25%< or =FA1/FA2/50% with WMT involvement; pattern 4, FA1/FA2<25% with WMT destruction.

CONCLUSIONSDTI allows for visualization of WMT and benefits surgical planning for patients with intrinsic brain tumor. There is a positive relationship between the bilateral FA ratio (FA1/FA2) variation and WMT alterations resulting from the tumor.

Adult ; Brain ; diagnostic imaging ; Brain Neoplasms ; diagnosis ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Glioblastoma ; diagnosis ; Glioma ; diagnosis ; Humans ; Male ; Middle Aged ; Nerve Fibers ; diagnostic imaging ; Neural Pathways ; radiation effects ; Preoperative Care ; Radiography ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVETo explore the effect of hypoxia on the fibrosis of corpus cavernosum smooth muscle cells (CCSMC) in SD rats.

METHODSCCSMCs were cultured in vitro, identified by immunohistochemistry, and then exposed to hypoxia at the concentration of 1% O2 for 12, 24, 48 and 72 hours. Those exposed to normal oxygen concentration for the corresponding lengths of time were used as the control. The relative expressions of TGF-131, type I collagen and type DI collagen were determined by RT-PCR.

RESULTSThe in vitro cultured CCSMCs grew well, and the anti-a-smooth muscle actin monoclonal antibodies were positive on immunohistochemical staining. The relative expression levels of TGF-beta1, type I collagen and type mI collagen were positively correlated with the time of hypoxia interference within 48 hours, and did not increase further with prolonged exposure.

CONCLUSIONWhen exposed to hypoxia, the relative expressions of TGF-beta1, type I collagen and type mI collagen in the CCSMCs of SD rats increased with the length of time, and reached the peak at 48 hours. Hypoxia can cause fibrosis of CCSMCs in SD rats.

Animals ; Cell Hypoxia ; Cells, Cultured ; Collagen Type I ; metabolism ; Extracellular Matrix ; Fibrosis ; Male ; Muscle, Smooth ; cytology ; pathology ; Oxygen ; metabolism ; Penis ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo assess the value of white matter fiber tractography based on magnetic resonance diffusion-tensor imaging (MR-DTI), a new noninvasive technique that helps estimate the structural connectivity of the brain, in understanding the anatomy of the white matter fiber tract.

METHODSSix consecutive volunteers received MR-DTI examination using a GE 1.5T Sign CV/I whole-body MR system and version 2.0 Functool software for image processing. The protocol included T1WI for localization and single-shot echo-planner diffusion tensor imaging of the entire brain in axial, sagittal, and /or coronal sections.

RESULTSWhite matter fiber tracts imaged by MR-DTI were consistent with known white matter fiber anatomy. The major fiber tracts were well observed in all the cases. The tracts visualized included the corpus callosum, anterior commissure, uncinate fasciculus superior and inferior longitudinal fascicules, inferior occipito-frontal fasciculus, internal capsule and corona radiate.

CONCLUSIONA set of detailed white matter fiber anatomy maps of the normal brain is obtained by means of single-shot echo-planner diffusion tensor imaging at high resolution. This technique allows rapid visualization of the white matter tracts in vivo, and provides a new reliable means for studying fiber connectivity in the brain.

Adult ; Brain ; anatomy & histology ; Diffusion Tensor Imaging ; methods ; Female ; Humans ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional ; methods ; Male ; Middle Aged ; Young Adult