OBJECTIVETo observe the effect of tetrandine on gene expression of collagen type I, collagen type III, transformation growth factor-beta1 and to investigate the inhibitory effect of tetrandine on the scar tissue hyperplasia in rabbits' ears.

METHODSAfter the scar model was formed on the rabbits' ears, the rabbits were divided into 4 groups to receive intro-lesion injection with saline, or prednisolone (Pre) or tetrandrine in low concentration (L-Tet, 1.0 mg/ml) or tetrandrine in high concentration (H-Tet, 7.5 mg/ml). The morphological changes of scar tissue were observed. The changes of fibroblasts quantity and collagen expression were observed with HE and Masson staining. Immunohistochemical study was used to observe the expression level of collagen type I and collagen type III and TGF-beta1. Collagen type I and collagen type III and TGF-beta1, and signal factor Smad 3 mRNA were detected with RT-PCR.

RESULTS(1) 24 days after injury, all the wounds healed completely with formation of red, tough and hypertrophic scar. HE and Masson staining showed significant increase of fibroblasts and collagen density with irregularly arrangement. (2) Compared with that in saline group, the scar in other groups became softer, lighter and thinner, especially in H-Tet group. (3) HE and Masson staining shows the scar in Tet and Pre groups contained less fibroblasts and lower collagen dentsity with comparatively regular arrangement than that in saline group (P < 0.01), especially in H-Tet group. (4) According to the immunohistochemical study, the expression of collage type I and III and TGF-beta was positive in all the groups, but the positive rate and the ratio of collagen density I to III decreased in the order of saline, L-Tet, H-Tet and Pre groups (P < 0.01). (5) PT-PCR detection results showed that the amplification bands brightness of collagen type I and III and TGF-beta1 and signal molecular Smad 3 mRNA in scar tissue were obviously different. Compared with that in saline group, the expression of collagen type I and III and TGF-beta1 and Smad 3 mRNA decreased in Tet and Pre groups (P < 0.01). H-Tet group showed the most obvious reduce in the expression of type I collagen and TGF-beta1 and Smad 3 mRNA. Conclusions Tetrandine can significantly suppress the expression of collagen type I and collagen type III and TGF-beta1 on hypertrophic scar of rabbit ears, and reduce signal factor Smad 3 mRNA' s expression. It may be one of the important mechanism for its inhibitory effect on scar hyperplasia.

Animals ; Benzylisoquinolines ; pharmacology ; Cicatrix, Hypertrophic ; drug therapy ; genetics ; pathology ; Collagen Type I ; genetics ; metabolism ; Collagen Type III ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Ear ; Fibroblasts ; Gene Expression ; Male ; RNA, Messenger ; metabolism ; Rabbits ; Smad3 Protein ; genetics ; metabolism ; Transforming Growth Factor beta1 ; genetics ; metabolism

Adolescent ; Adult ; Aged ; Child ; Endoscopes ; Female ; Humans ; Male ; Middle Aged ; Otorhinolaryngologic Surgical Procedures ; Smell ; Turbinates ; surgery ; Young Adult

Nitrites play multiple characteristic functions in invasion and metastasis of hepatic cancer cells, but the exact mechanism is not yet known. Cancer cells can maintain the malignant characteristics via clearance of excess mitochondria by mitophagy. The purpose of this article was to determine the roles of nitrite, reactive oxygen species (ROS) and hypoxia inducing factor 1 alpha (HIF-1 α) in mitophagy of hepatic cancer cells. After exposure of human hepatocellular carcinoma SMMC-7721 cells to a serial concentrations of sodium nitrite for 24 h under normal oxygen, the maximal cell vitality was increased by 16 mg x (-1) sodium nitrite. In addition, the potentials of migration and invasion for SMMC-7721 cells were increased significantly at the same time. Furthermore, sodium nitrite exposure displayed an increase of stress fibers, lamellipodum and perinuclear mitochondrial distribution by cell staining with Actin-Tracker Green and Mito-Tracker Red, which was reversed by N-acetylcysteine (NAC, a reactive oxygen scavenger). DCFH-DA staining with fluorescent microscopy showed that the intracellular level of ROS concentration was increased by the sodium nitrite treatment. LC3 immunostaining and Western blot results showed that sodium nitrite enhanced cell autophagy flux. Under the transmission electron microscopy (TEM), more autolysosomes formed after sodium nitrite treatment and NAC could prevent autophagosome degradation. RT-PCR results indicated that the expression levels of COX I and COXIV mRNA were decreased significantly after sodium nitrite treatment. Meanwhile, laser scanning confocal microscopy showed that sodium nitrite significantly reduced mitochondrial mass detected by Mito-Tracker Green staining. The expression levels of HIF-1α, Beclin-1 and Bnip3 (mitophagy marker molecular) increased remarkably after sodium nitrite treatment, which were reversed by NAC. Our results demonstrated that sodium nitrite (16 mg x L(-1)) increased the potentials of invasion and migration of hepatic cancer SMMC-7721 cells through induction of ROS and HIF-1α mediated mitophagy.
Acetylcysteine ; pharmacology ; Autophagy ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Movement ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Liver Neoplasms ; pathology ; Mitochondrial Degradation ; Neoplasm Invasiveness ; Nitrites ; metabolism ; Reactive Oxygen Species ; metabolism ; Sodium Nitrite ; pharmacology

0.05).The positive rate of Uu in biovar 2 show a significant difference(P0.05).(2)The fallopian tubes infected by biovar2 have a high rate(90%)of ciliary adhesion and exuviation.While there is a low rate(10%)for biovarl with ciliary adhesion and exuviation.There was significant difference between the two groups of Uu (P

The influence of separating effect of different chromatographic conditions of mouse gut flora by high performance ion exchange chromatography analysis was studied. The optimum chromatographic conditions for separating gut bacteria were determined. The sample was applied to the chromatography column packed with Toyopearl SuperQ-650c anion resin, equilibrated with 0.02mol/L piperazin-hydrochloric acid buffer (pH 8.0), and elution salt 1mol/L NaCl, eluted with the gradient of 0-50% NaCl/ 80 min, then 50%~75% NaCl/ 25 min at the flow rate 1ml/min, and injecting volume was 1ml.Under these conditions, intestinal flora were separated into several fractions. The establishment of HPLC analysis method will lay a foundation of further research on the components of mouse gut flora and their dynamic changes.

In gene manipulation, different selectable markers with various linkers are necessary. In order to get selectable markers directly, we constructed from pBlueScript SK(-) a series of particular plasmids, pSKsymKm, pSKsymBle, pSKsymEry, pSKsymHyg and pSKsymGm, each contains Kanamycin, Bleomycin, Erythromycin, Hygromycin or Gentamycin resistance cassette. By restriction enzyme digestion and gel extraction, any of five antibiotic resistance genes with specific ends can be conveniently obtained.

AIM:To observe related biological parameters of 3 minutes dark-room provocative test in patients with laser peripheral iridectomy(LPI) in the fellow eyes of acute primary angle-closure (APAC) by ultrasound biomicroscopy (UBM).To explore the risk factors in primary angle closure suspect(PACS) patients with progressive angle closure after LPI.METHODS: Seventy-eight eyes of APAC patients without peripheral anterior synechia were selected.Each eye underwent 3 minutes dark-room provocative test after LPI.Anterior segment parameters, including anterior chamber depth (ACD), anterior chamber angle open distance500 (AOD500), peripheral iris thickness (PIT), iris convex (IC), the position of iris insertion and trabecular-ciliary process distance (TCPD), and the number of positional angle closure(NPAC) were observed and analyzed by statistic methods.RESULTS:Patients with APAC were examined by UBM after LPI and 26 eyes(33%) occurs at least one positional angle closure,19 eyes(24%)were positive in 3 minutes dark-room provocative test among them.It occurs a positive relationship between the elevation intraocular pressure and the number of positional angle closure in dark-room provocative test(r=0.84, P<0.01).AOD500, IT and IC were significantly changed from normal light to darkroom between positional angle closure positive group and positional angle closure negative group(all P<0.01).In single factor analysis, AOD500(P=0.003), IT(P=0.012), IC(P=0.043), TPCD(P=0.015), the position of iris insertion(P=0.024) were correlative factors of positive results.In multiple-factor analysis, only IT(P=0.011), TPCD(P=0.009), iris root attachment points(P=0.02) were independent risk factors of positive results.CONCLUSION:A certain proportion of patients with PACS after LPI appeared positional angle closure in a dark room.Peripheral iris hypertrophy, anterior displacement of the ciliary body and iris root attachment points are vital risk factors.Long-term follow-up study and intervention treatment are required in these patients after LPI.

Nitrites play multiple characteristic functions in invasion and metastasis of hepatic cancer cells, but the exact mechanism is not yet known. Cancer cells can maintain the malignant characteristics via clearance of excess mitochondria by mitophagy. The purpose of this article was to determine the roles of nitrite, reactive oxygen species (ROS) and hypoxia inducing factor 1 alpha (HIF-1 α) in mitophagy of hepatic cancer cells. After exposure of human hepatocellular carcinoma SMMC-7721 cells to a serial concentrations of sodium nitrite for 24 h under normal oxygen, the maximal cell vitality was increased by 16 mg x (-1) sodium nitrite. In addition, the potentials of migration and invasion for SMMC-7721 cells were increased significantly at the same time. Furthermore, sodium nitrite exposure displayed an increase of stress fibers, lamellipodum and perinuclear mitochondrial distribution by cell staining with Actin-Tracker Green and Mito-Tracker Red, which was reversed by N-acetylcysteine (NAC, a reactive oxygen scavenger). DCFH-DA staining with fluorescent microscopy showed that the intracellular level of ROS concentration was increased by the sodium nitrite treatment. LC3 immunostaining and Western blot results showed that sodium nitrite enhanced cell autophagy flux. Under the transmission electron microscopy (TEM), more autolysosomes formed after sodium nitrite treatment and NAC could prevent autophagosome degradation. RT-PCR results indicated that the expression levels of COX I and COXIV mRNA were decreased significantly after sodium nitrite treatment. Meanwhile, laser scanning confocal microscopy showed that sodium nitrite significantly reduced mitochondrial mass detected by Mito-Tracker Green staining. The expression levels of HIF-1α, Beclin-1 and Bnip3 (mitophagy marker molecular) increased remarkably after sodium nitrite treatment, which were reversed by NAC. Our results demonstrated that sodium nitrite (16 mg x L(-1)) increased the potentials of invasion and migration of hepatic cancer SMMC-7721 cells through induction of ROS and HIF-1α mediated mitophagy.

The role of flavonoids of Echinps latifolius (FELT) in Wnt signaling was investigated in adjuvant arthritis (AA) rats. The therapeutic effects of FELT on AA rats were detected by rat arthritis score and MTT. The effect of FELT gavage treatment on the Wnt signaling key gene β-catenin, C-myc and cyclin D1 in synovium from AA rats was detected by Real-time qPCR, and the effects of FELT gavage treatment on the upstream negative regulation gene SFRP 1,2,4,5 in synovium from AA rats were detected by Real-time qPCR. The results showed that FELT gavage treatment significantly inhibited arthritis score and MTT values in AA rats, significantly inhibited the expression of the Wnt signaling gene β-catenin, C-myc and cyclin D1, significantly up-regulated the expression of the up- stream negative regulation gene SFRP 1,2,4. FELT has a better therapeutic effect for AA rats.
Animals ; Arthritis, Experimental ; drug therapy ; genetics ; metabolism ; Asteraceae ; chemistry ; Disease Models, Animal ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Flavonoids ; administration & dosage ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Male ; Membrane Proteins ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Synovial Membrane ; drug effects ; metabolism ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; metabolism

The clinical efficacy of acupuncture and moxibustion for post-stroke constipation was systematically reviewed. By computerized and manual retrieval of clinical research literature regarding acupuncture and moxibustion for post-stroke constipation, the randomized control trials (RCTs) that met the inclusive criteria were collected. Cochrane systematic review method was used and Revmen 5.2 software was adopted to perform this Meta analysis. Totally 8 articles were included, involving 610 cases of post-stroke constipation. As a result, the total effective rate and cured rate of acupuncture and moxibustion for post-stroke constipation were significantly superior to those of the control group [total effective rate: OR = 2.10, 95% CI (1.25, 3.54), Z = 2.78, P = 0.005; cured rate: OR = 2.37, 95% CI (1.57, 3.58), Z = 4.10, P < 0.0001]. This result indicated that acupuncture was effective for post-stroke constipation and had some advantages compared with other therapies. But the quality of included RCTs was low, and high-quality, large-sample and multi-center RCTs were needed to perform further verification.
Acupuncture Therapy ; Constipation ; etiology ; therapy ; Humans ; Moxibustion ; Randomized Controlled Trials as Topic ; Stroke ; complications ; Treatment Outcome