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1.
Chinese Pharmacological Bulletin ; (12): 631-637, 2023.
Article in Chinese | WPRIM | ID: wpr-1013929

ABSTRACT

Aim To investigate the effect of RORα antagonist T0901317 promoting EMT (epithelial-mesenchymal transition) of human gastric cancer MGC803 cells by RORα/β-catenin signal. Methods Cell proliferation was detected by MTT. Cell migration and invasion were detected by cell scratch and Transwell assay respectively. RORα/β-catenin signaling molecules were detected by Western blot and immunofluorescence. RORα binding to β-catenin protein was detected by immunoprecipitation. Results MTT assay showed that the proliferation ability of T0901317 cells increased in a time-dependent manner compared with MGC803 cells (P < 0. 05). Cell scratches and Transwell experiments showed that the migration and invasion ability of T0901317 cells were significantly enhanced compared with MGC803 cells (P < 0. 05). Western blot analysis showed that RORα protein was significantly down-regulated after T0901317 compared with untreated group (P < 0. 05), and total β-catenin protein and nuclear β-catenin in MGC803 cells were up-regulated after T0901317 (P < 0. 05). Compared with the control group, RORα protein binding to β-catenin protein significantly decreased after T0901317 treatment (P < 0. 05). Compared with MGC803 cells treated with T0901317, the long spindle cells increased and the heteromorphism was more obvious. T0901317 significantly up-regulated the expression of Rac1, TGFβ1 and Vimentin in MGC803 cells (P < 0. 05), and inhibited the expression of E-cadherin (P < 0. 05). Conclusion T0901317 can promote the proliferation, migration, invasion and EMT in human gastric cancer MGC803 cells by RORα/β-catenin signal.

2.
China Journal of Chinese Materia Medica ; (24): 1892-1898, 2023.
Article in Chinese | WPRIM | ID: wpr-981408

ABSTRACT

The present study aimed to explore the chemical constituents from the stems and leaves of Cephalotaxus fortunei. Seven lignans were isolated from the 75% ethanol extract of C. fortunei by various chromatographic methods, including silica gel, ODS column chromatography, and HPLC. The structures of the isolated compounds were elucidated according to physicochemical properties and spectral data. Compound 1 is a new lignan named cephalignan A. The known compounds were identified as 8-hydroxy-conidendrine(2), isolariciresinol(3), leptolepisol D(4), diarctigenin(5), dihydrodehydrodiconiferyl alcohol 9'-O-β-D-glucopyranoside(6), and dihydrodehydrodiconiferyl alcohol 4-O-β-D-glucopyranoside(7). Compounds 2 and 5 were isolated from the Cephalotaxus plant for the first time.


Subject(s)
Cephalotaxus , Lignans/analysis , Plant Leaves/chemistry , Ethanol , Chromatography, High Pressure Liquid
3.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 78-85, 2022.
Article in Chinese | WPRIM | ID: wpr-940695

ABSTRACT

ObjectiveTo investigate the possible mechanism of Bushen Zhuyun prescription (BSZYP) to reduce the level of ovarian apoptosis in Brown Norway (BN) rats with luteal phase deficiency (LPD). MethodFifty SPF female BN rats were randomly divided into a model group, a dydrogesterone group (0.002 g·kg-1), and a low (4.5 g·kg-1), a medium (9 g·kg-1), and a high-dose (18 g·kg-1) BSZYP groups, with ten rats in each group. The rats were administrated with corresponding drugs by gavage, once a day for three estrus cycle. Western blot was used to detected the protein expression levels of c-Jun NH2 terminal kinase (JNK), extracellular signal-regulated kinase (ERK), phosphorylated-ERK (p-ERK), phosphorylated-JNK (p-JNK), p38 mitogen-activated protein kinase (p38 MAPK), phosphorylated-p38 MAPK (p-p38 MAPK ), B-cell lymphoma (Bcl-2), and Bcl-2 associated X protein (Bax) in ovary. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression levels of ERK, JNK, p38 MAPK, Bax, and Bcl-2 in ovary. Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum progesterone (P) and estradiol (E2) levels. Hematoxylin-eosin (HE) staining was used to observe the ovarian tissue morphology. ResultCompared with the model group, the recovery of estrus cycle of rats in all BSZYP groups had statistical significance after 1-circle administration (P<0.05). The ovarian tissue morphology in the low-dose BSZYP group was improved, and that in the medium and high-dose BSZYP groups was significantly improved with clear follicle, less vesicular follicle and atretic follicle, and more granular layers. The number of luteum, especially the fresh luteum, in the medium and high-dose groups was increased with smooth edge and large volume. The mRNA expression level of Bcl-2 was increased in all-dose BSZYP groups, while the mRNA expression level of Bax was significantly decreased in all-dose BSZYP group (P<0.05, P<0.01). The mRNA expression levels of JNK and p38 MAPK were significantly decreased in the high-dose BSZYP group (P<0.01), and the mRNA expression levels of ERK were increased in the low and medium-dose BSZYP groups (P<0.05). The protein expression level of Bcl-2 was significantly increased in the medium and high-dose BSZYP groups (P<0.01), and the protein expression level of Bax was significantly decreased in the all-dose BSZYP groups (P<0.01). No significant difference was observed in the protein expressions of JNK, ERK, and p38 MAPK in the BSZYP groups. The protein expression levels of p-ERK in the ovarian tissues of rats were significantly inoreased in the medium and high-dose BSZYP group (P<0.01), p-JNK, and p-p38 MAPK in the ovarian tissues of rats were significantly decreased in the medium and high-dose BSZYP group (P<0.01). The level of E2 was increased in all-dose BSZYP groups (P<0.05, P<0.01), and the level of P in the medium-dose BSZYP group was increased (P<0.05). ConclusionBSZYP improved the serum sex hormones, restored the estrous cycle, reduced atretic follicle and vesiculation, and maintained luteal morphology and function of BN rats, so as to improve luteal function and treat luteal phase deficiency. The mechanism of BSZYP may be related to reduce the level of ovarian tissue apoptosis in BN rats by regulating MAPKs signaling pathway.

4.
Chinese Journal of Pathology ; (12): 326-331, 2022.
Article in Chinese | WPRIM | ID: wpr-935534

ABSTRACT

Objective: To investigate the clinicopathological and molecular characteristics of hepatic fibrinogen storage disease (FSD) in children. Methods: The clinical, histopathologic, immunophenotypic, ultrastructural and gene sequencing data of 4 FSD cases were collected from September 2019 to January 2021 in the Children's Hospital of Fudan University, Shanghai, China. Retrospective analysis and literature review were conducted. Results: There were 4 cases of FSD, 3 males and 1 female, aged 3 years and 3 months to 6 years (median age, 3 years and 4 months). The clinical manifestations were abnormal liver function and abnormal blood coagulation function, for which 2 cases had family genetic history. Liver biopsies revealed that, besides liver steatosis, fibrosis and inflammation, there were single or multiple eosinophilic inclusion bodies of various sizes and surrounding transparent pale halo in hepatocytes. Immunohistochemistry showed that the inclusion bodies were positive for anti-fibrinogen. Under the electron microscope, they corresponded to the dilated cisternae of the rough endoplasmic reticulum, which were occupied by compactly packed tubular structures and arranged into a fingerprint-like pattern with curved bundles. Gene sequencing revealed that the 2 cases of FGG mutation were located in exon 8 c.1106A>G (p.His369Arg) and c.905T>C (p.Leu302Pro), and 1 case was located in exon 9 c.1201C>T (p.Arg401Trp). No pathogenic variant was detected in the other case. Conclusions: FSD is a rare genetic metabolic disease and clinically manifests as abnormal liver function with hypofibrinogenemia. In the background of liver steatosis, fibrosis and inflammation, there are eosinophilic inclusions with pale halo in the hepatocytic cytoplasm, which can be identified by anti-fibrinogen immunohistochemical staining. The fingerprint-like structures under electron microscope are helpful for the diagnosis, while FGG sequencing detects the pathogenic mutation of exon 8 or 9 that can clearly explain the phenotype. However, the diagnosis of FSD cannot be completely ruled out if the relevant mutations are not detected.


Subject(s)
Child , Child, Preschool , Female , Humans , Male , China , Fibrinogen/chemistry , Liver/pathology , Liver Diseases/pathology , Metabolic Diseases/pathology , Retrospective Studies
5.
China Journal of Chinese Materia Medica ; (24): 3797-3805, 2021.
Article in Chinese | WPRIM | ID: wpr-888101

ABSTRACT

Terpenes are the largest group of natural products and contain the widest assortment of structural types. Terpene cyclization is also the most complex reaction found in nature. For a long time, terpenoids with diverse structures have attracted natural product chemists to explore their biosynthesis mechanism. Such a large number of terpene skeletons are catalyzed by enzymes called terpene synthase. Sesquiterpene synthase is a kind of terpene synthase, which can catalyze the cyclization of linear precursor farnesyl pyrophosphate(FPP) to sesquiterpene skeletons. Sesquiterpene synthase cyclize a single precursor FPP into many sesquiterpene skeletons. With the continuous discovery of sesquiterpene synthase, the cyclization mechanism of sesquiterpene synthase has been studied deeply. In recent years, with the development and improvement of isotope labeling of substrate FPP and structural analysis of sesquiterpene synthase, the structure and cyclization mechanism of sesquiterpene synthase have been studied more systematically and accurately. In this review, we reviewed the progress of the research methods on the mechanism of sesquiterpene cyclization by substrate isotope labeling and protein structure, as well as the summary and prospect of sesquiterpene synthase research.


Subject(s)
Cyclization , Sesquiterpenes , Terpenes
6.
China Journal of Chinese Materia Medica ; (24): 6493-6501, 2021.
Article in Chinese | WPRIM | ID: wpr-921809

ABSTRACT

The present study investigated the effect of Modified Dihuang Decoction in improving ovarian reserve in mice through the Bcl-2-related mitochondrial apoptosis pathway. Forty-eight adult female BALB/c mice were randomly divided into the following six groups with eight mice in each group: a blank group, a model group, a femoston group(three cycles of treatment with 0.13 mg·kg~(-1) estradiol tablets for 2 days and 1.43 mg·kg~(-1) estradiol and dydrogesterone tablets for 3 days), and high(64.74 g·kg~(-1))-, medium(43.16 g·kg~(-1))-, and low-dose(21.58 g·kg~(-1)) Modified Dihuang Decoction groups. Mice in other groups except the blank group received a single intraperitoneal injection of 12 mg·kg~(-1) cyclophosphamide and 1.2 mg·kg~(-1) busulfan to induce a model of diminished ovarian reserve(DOR), while those in the blank group received an equal volume of normal saline. Mice were treated with corresponding drugs for 15 d from the 36 th day, once per day, and the mice in the blank group and the model group were treated with an equal volume of normal saline. The general condition and oestrous cycle were observed. The serum hormone levels were detected with the enzyme-linked immunosorbent assay(ELISA). The morphological changes of ovaries were observed by HE staining. Western blot was used to detect the protein expression of cysteinyl aspartate specific proteinase-9(caspase-9), cleaved caspase-3, Bcl-2 associated X protein(Bax), Bcl-2, superoxide dismutase-2(SOD-2), and glutathione peroxidase-1(GPx-1). The mRNA expression of Bax and Bcl-2 was detected by real-time fluorescence-based quantitative polymerase chain reaction(real-time PCR). The results showed that compared with the blank group, the model group showed body weight loss, disordered oestrous cycle, elevated serum levels of follicle-stimulating hormone(FSH) and luteinizing hormone(LH), reduced serum levels of estradiol(E_2), anti-mullerian hormone(AMH), and inhibin B(INHB), the declining number of ovarian follicles and granulosa layers, increased number of atretic follicles, up-regulated protein expression of caspase-9, cleaved caspase-3, and Bax and Bax mRNA expression in ovaries, and down-regulated protein expression of Bcl-2, SOD-2 and GPx-1, and Bcl-2 mRNA expression. Compared with the model group, the Modified Dihuang Decoction groups displayed restored body weight and oestrous cycle, decreased serum levels of FSH and LH, elevated serum levels of E_2, AMH, and INHB, increased number of ovarian follicles, thickened granulosa layers, and declining number of atretic follicles. Additionally, the protein expression of caspase-9, cleaved caspase-3, and Bax, and Bax mRNA expression was down-regulated, and the protein expression of Bcl-2, SOD-2, and GPx-1, and Bcl-2 mRNA expression was up-regulated. The results suggest that Modified Dihuang Decoction can regulate endocrine hormone, promote follicle growth and improve ovarian reserve by enhancing ovarian anti-oxidant capacity, inhibiting the Bcl-2-related mitochondrial apoptosis pathway, and further inhibiting cell apoptosis.


Subject(s)
Animals , Female , Mice , Apoptosis , Mice, Inbred BALB C , Ovarian Follicle , Ovarian Reserve , Ovary
7.
Chinese Journal of Rehabilitation Theory and Practice ; (12): 423-431, 2020.
Article in Chinese | WPRIM | ID: wpr-905454

ABSTRACT

Objective:To explore the effect of Tuina of Three Handing-Three Points on the recovery of motor function, the expression of neuregulin (NRG) 1 and human epidermal growth factor receptor (ErbB) 2 in the injured point of sciatic nerve and L4-6 spinal cord, and the morphological change of myelin sheath at the injured point of sciatic nerve of rats. Methods:A total of 76 male Sprague-Dawley rats were randomly divided into normal group, sham operation group, model group and Tuina group with 19 rats in each group. The right side sciatic nerve was clamped to make model in the model group and Tuina group. The sham operation group exposed sciatic nerve only. Tuina group received Tuina on Yinmen (BL37), Chengshan (BL57) and Yanglingquan (GB34) with dialing, plucking and kneading using Tuina technique simulator. All of them were tested with Oblique Plate Test before modeling, seven days and 28 days after modeling. Western blotting was used to detect the protein expression of NRG1 and ErbB2 in the injured point of sciatic nerve and L4-6 spinal cord. The change of myelin sheath at the sciatic nerve injury point was observed and analyzed by transmission electron microscope. Results:Seven days and 28 days after modeling, the scores of Oblique Plate Test were lower in the model group and Tuina group than in the normal group and the sham operation group (P < 0.05); 28 days after modeling, the scores was higher in Tuina group than in the model group (P < 0.05). At the sciatic nerve injury point, three days after modeling, the expression of NRG1 and ErbB2 was higher in the model group and Tuina group than in the normal group and the sham operation group (P < 0.05); seven days and 28 days after modeling, there was no significant difference in NRG1 among groups (P > 0.05); 28 days after modeling, the expression of ErbB2 was higher in the model group and Tuina group than in the normal group and the sham operation group (P < 0.05). In L4-6 spinal cord, three days after modeling, the expression of NRG1 and ErbB2 was higher in the model group and Tuina group than in the normal group and sham operation group (P < 0.05); seven days after modeling, the expression of NRG1 was higher in the model group and Tuina group than in the sham operation group (P < 0.05), and the expression of ErbB2 was higher in the model group and Tuina group than in the normal group and the sham operation group (P < 0.05); 28 days after modeling, the expression of NRG1 was higher in Tuina group than in the model group (P < 0.05), and there was no significant difference in ErbB2 among groups (P > 0.05). The electron microscope showed that, 28 days after modeling, the myelin sheath collapsed seriously in the model group; while the ultrastructure of the nerve injury point improved, and the myelin sheath of the nerve fiber was relatively intact in Tuina group; the g-ratio value was lower in the model group than in the sham operation group (P < 0.05), the g-ratio value was higher in Tuina group than in the model group (P < 0.05), and no difference was found in g-ratio value between Tuina group and sham operation group (P > 0.05). Conclusion:Three Handing-Three Points could improve the motor function of hind limbs in rats with sciatic nerve injury, which may be related to the adjustment of NRG1 and ErbB2 in the sciatic nerve and spinal cord, to maintain normal myelin sheath structure.

8.
Chinese Journal of Rehabilitation Theory and Practice ; (12): 964-968, 2020.
Article in Chinese | WPRIM | ID: wpr-905421

ABSTRACT

Objective:To observe the effect of Dredging Correcting Manipulation on cblC methylmalonic aciduria (MMA). Methods:From October, 2017 to October, 2018, 72 children with cblC MMA combined with growth retardation were divided into control group (n = 36) and experimental group (n = 36) according to the consent of their parents. The control group accepted routine medicine, and the experimental group received Dredging Correcting Manipulation in addition. The Griffiths Development Scale-Chinese version (GDS-C) was used to evaluate the two groups before and after treatment. At the same time, body length, body mass and head circumference were measured. Results:Six cases in the control group and five cases in the experimental group were dropped out. There was no significant difference in the development quotients of GDS-C in gross movement, personal and social, hearing and speech, hand-eye coordination, operation and total quotient between two groups before treatment (P > 0.05). After treatment, all the development quotients increased in both groups (t > 6.110, P < 0.001), and the development quotients of GDS-C in gross movement, personal and social, hand-eye coordination and total quotient were higher in the experimental group than in the control group (t > 2.154, P < 0.05), as well as the body length (t = 2.027, P < 0.05). Conclusion:Dredging Correcting Manipulation can promote the neuropsychological and physical development of children with cblC MMA combined with retardation.

9.
Chinese Journal of Rehabilitation Theory and Practice ; (12): 881-884, 2020.
Article in Chinese | WPRIM | ID: wpr-905406

ABSTRACT

Intellectual development disorders are a group of etiologically diverse conditions originating during the developmental period characterized by significantly below average intellectual functioning and adaptive behavior. It is important to combine the personalized intervention with games and activities of daily living in community and family. Moreover, early intervention methods such as multi-sensory stimulation, activity observation training, goals-activity-motor enrichment are needed to improve their cognitive function; special education, intervention of behavioral, medicine and training of social skills, self-esteem and emotional quotient, to improve the outcome of children with intellectual development disorder.

10.
Chinese Traditional and Herbal Drugs ; (24): 3720-3727, 2017.
Article in Chinese | WPRIM | ID: wpr-852518

ABSTRACT

Objective To optimize the preparation process of honokiol long-circulating liposomes (HLCL) and study the in vitro and in vivo release. Methods An orthogonal experiment was designed to optimize the composition of HLCL using entrapment efficiency as evaluation indicator. The liposome surface morphology was observed by transmission electron microscope (TEM), and the liposome release in vitro was studied by dialysis method. The concentration of honokiol in rat plasma was determined by the established LC-MS/MS method, and the differences in pharmacokinetic parameters were compared after honokiol and HLCL (20 mg/kg) were orally administered to SD male rats, respectively. Results The optimal composition of HLCL was 8:1:1 for soya phosphatidyl choline-cholesterol-mPEG2000-DSPE, and 1:10 for honokiol-liposome materials with the ultrasonic time of 12 min. Under the optimized conditions, HLCL was sphere with mean particle size of 121.5 nm and mean Zeta potential of -30.8 mV, the encapsulation efficiency and drug-loading content was 84.7% and 10.4%, respectively. In vitro release results showed that the liposomes could be gently and slowly release with the 24 h cumulative release rate at pH 1.2 and pH 6.9 dissolve medium of 80% and 71%, respectively. Based on the pharmacokinetic results, Cmax, tmax, and t1/2 were (23.29 ± 11.76) ng/mL, (0.13 ± 0.05) h and (10.59 ± 5.72) h for HLCL, and (79.34 ± 56.32) ng/mL, (0.30 ± 0.07) h and (4.44 ± 3.14) h for honokiol, respectively. There was no significant difference about the AUC0-∞ following oral administration of honokiol and HLCL at isodose honokiol (20 mg/kg). Conclusion Compared with honokiol, HLCL was released gently and slowly in vitro, absorpted rapidly and eliminated slowly in vivo.

11.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 20-24, 2017.
Article in Chinese | WPRIM | ID: wpr-238391

ABSTRACT

The risk factors of high trait anger of juvenile offenders were explored through question naire study in a youth correctional facility of Hubei province,China.A total of 1090 juvenile offenders in Hubei province were investigated by self-compiled social-demographic questionnaire,Childhood Trauma Questionnaire (CTQ),and State-Trait Anger Expression Inventory-Ⅱ (STAXI-Ⅱ).The risk factors were analyzed by chi-square tests,correlation analysis,and binary logistic regression analysis with SPSS 19.0.A total of 1082 copies of valid questionnaires were collected.High trait anger group (n=316) was defined as those who scored in the upper 27th percentile of STAXI-Ⅱ trait anger scale (TAS),and the rest were defined as low trait anger group (n=766).The risk factors associated with high level of trait anger included:childhood emotional abuse,childhood sexual abuse,step family,frequent drug abuse,and frequent internet using (P<0.05 or P<0.01).Birth sequence,number of sibling,ranking in the family,identity of the main care-taker,the education level of care-taker,educational style of care-taker,family income,relationship between parents,social atmosphere of local area,frequent drinking,and frequent smoking did not predict to high level of trait anger (P>0.05).It was suggested that traumatic experience in childhood and unhealthy life style may significantly increase the level of trait anger in adulthood.The risk factors of high trait anger and their effects should be taken into consideration seriously.

12.
Chinese Traditional and Herbal Drugs ; (24): 2701-2706, 2016.
Article in Chinese | WPRIM | ID: wpr-853373

ABSTRACT

Objective: To study the dynamic processes of in vivo metabolism of multicomponent in the decoction of Angelicae Sinensis Radix (ASR), select the quality control components, look for a new quality evaluation method for Chinese materia medica (CMM). Methods: The blood was taken from mesenteric vein and abdominal aorta. By analyzing HPLC fingerprints of intestinal metabolism and liver metabolism, we could infer multicomponent changes in the metabolic process of water extract from ASR. Results: Totally 15 components were basically stable in the digestive juice, four of them may be metabolized by intestine, while seven of them were metabolized by liver. And we found one new component was the metabolite of liver, seven new components were the metabolites of intestine, but five of them were metabolized by liver and could not be absorbed into blood. Finally, we identified that four prototype components of water extract from ASR and three metabolites could be absorbed into blood. Conclusion: Based on the multicomponent metabolic profiling of ASR, we initially select the four prototype components including senkyunolide I as the quality control component, and we finally establish a quality evaluation method for "correlation in quality and metabolism".

13.
Journal of Southern Medical University ; (12): 256-260, 2016.
Article in Chinese | WPRIM | ID: wpr-273778

ABSTRACT

<p><b>OBJECTIVE</b>To elucidate the molecular and electrophysiological mechanisms of Brugada syndrome through functional analysis of a novel SCN5A gene mutation G1712C.</p><p><b>METHODS</b>A recombinant plasmid pRc<CMV-hH1 containing the mutant human cardiac sodium channel α subunit (hH1) cDNA was constructed using in vitro PCR-based site-directed mutagenesis technique. LipofectamineTM 3000 was used to transfect the plasmid DNA into HEK293 cell line to induce stable expression of Nachannel β1-subunit, and the positive colonies were selected by screening with G418.The standard liposome method was used to transiently transfect HEK293 cells with either the wild-type or mutant Nachannel subunits (hH1 and mhH1, respectively), and the macroscopic Nacurrents were recorded using whole-cell patch-clamp technique. Data acquisition and analysis, generation of voltage commands and curve fitting were accomplished with EPC-10, PatchMaster and IGOR Pro 6.0.</p><p><b>RESULTS</b>An HEK293 cell line that stably expressed Nachannel β1-subunit was successfully established. After transient transfection with the WT subunit, large Nacurrents were recorded from the stable β1-cell line. Transient transfection with the G1712C subunit, however, did not elicit a Nacurrent in the cells.</p><p><b>CONCLUSION</b>Compared with normal Nachannel, the wild-type channel exhibits a similar sodium current. The characteristic kinetics of sodium channel of WT-hH1 was identical to that in normal cardiac muscle cell, and the missense mutation (G1712C) in the P-loop region of the domain IV may have caused the failure of sodium channel expression.</p>


Subject(s)
Humans , Brugada Syndrome , Genetics , Genotype , HEK293 Cells , Mutagenesis, Site-Directed , Mutation , Genetics , Patch-Clamp Techniques , Polymerase Chain Reaction , Transfection
14.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 314-319, 2015.
Article in Chinese | WPRIM | ID: wpr-297433

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effect of berberine on lipid metabolism disorder and lipid deposition in liver cells of non-alcoholic fatty liver disease (NAFLD) rats induced by high fat diet.</p><p><b>METHODS</b>After one week adaptable feeding, 45 SPF level male SD rats were randomly divided into 3 groups, the normal control group, the model group, and the berberine group, 15 in each group. Except those in the normal control group, all rats were fed with high fat diet to prepare NAFLD model. As for rats in the berberine group, Berberine Hydrochloride was administered by gastrogavage. HE staining and oil red O staining were performed to identify the model after 8 weeks. Hepatocytes were isolated, and their activities and purities were tested by Typan blue staining and flow cytometry (FCM). Serum levels of TC, TG, HDL-C, and LDL-C were detected using automatic biochemical analyzer. mRNA expression levels of LXRα and FAS in liver cells were analyzed by Real-time quantitative polymerase chain reaction (PCR). Protein levels of LXRα and FAS in liver cells were examined by Western blot.</p><p><b>RESULTS</b>The NAFLD rat model was successfully established by high fat diet. The yields of purified liver cells in each rat were (6.0-7.5) x 10(8). The viability of isolated liver cells with purity over 90% (tested by FCM analysis) was higher than 95%. Compared with the normal control group,the expression of LXRα and FAS at mRNA and protein levels was higher in the model group (P < 0.01). Compared with the model group, the expression of LXRα and FAS at mRNA and protein levels was obviously down-regulated in the berberine group (P < 0.01).</p><p><b>CONCLUSIONS</b>LXRα/FAS signaling pathway was one of important signaling pathways of NAFLD lipid metabolism disorders. Berberine could recover hepatocyte fatty deposits in NAFLD rats by adjusting the LXR/FAS signaling pathway of hepatocytes, which might be one of important mechanisms for fighting against NAFLD.</p>


Subject(s)
Animals , Male , Rats , Berberine , Therapeutic Uses , Diet, High-Fat , Down-Regulation , Drugs, Chinese Herbal , Therapeutic Uses , Fatty Liver , Hepatocytes , Lipids , Non-alcoholic Fatty Liver Disease , Drug Therapy , RNA, Messenger , Rats, Sprague-Dawley , Signal Transduction
15.
China Journal of Chinese Materia Medica ; (24): 4027-4033, 2014.
Article in Chinese | WPRIM | ID: wpr-310947

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effects of soothing liver and invigorating spleen recipes on lipopolysaccharide(LPS) induced hepatocyte inflammation of rats and TLR4/p38MAPK signal pathway.</p><p><b>METHOD</b>The hepatocytes of SD rats were cultured and identified in vitro. The medicated serum of soothing liver and invigorating spleen recipes was prepared. The hepatocytes were treated with soothing liver and invigorating spleen recipes. Then Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expression in cultural supernatants were assayed by ELISA. The expressions of Toll-Like 4 (TLR4), p38 mitogen activated protein kinases (p38MAPK) and p-p38 mitogen-activated protein kinase (p-p38MAPK) were detected by Western blot.</p><p><b>RESULT</b>The rat medicated serum of soothing liver and invigorating spleen recipes was extracted for 2-3 mL. The purified rat hepatocytes were 1.5 x 10(8)-2.0 x 10(8). The cell viability was above 95% detected by Typan blue staining. The hepatocytes were identified by immumofluorescence assay. The detection of hepatocyte cultural supernatants: compared with that of the control group, IL-6 and TNF-α expression were increased in the LPS group (P < 0.01). While compared with that of the LPS group, the expressions of IL-6 and TNF-α were decreased after soothing liver and invigorating spleen recipes intervention (P < 0.01). The detection of hepatocyte proteins: compared with that of the control group, the protein expressions of p38MAPK, p-p38MAPK and TLR4 were all increased significantly in the LPS group (P < 0.01). Compared with that of the LPS group, the protein expressions of p38MAPK was decreased significantly in SB239063 group and it was also decreased in the soothing liver and invigorating spleen recipes group, but with no significant difference. Compared with that of the LPS group, p38MAPK expression was reduced significantly in the soothing liver and invigorating spleen recipes group and the SB239063 (p38MAPK pathway inhibitor) group (P < 0.01). TLR4 protein expression was decreased markedly in the soothing liver and invigorating spleen recipes group (P < 0.01) but had no difference between the SB239063 group and the LPS group.</p><p><b>CONCLUSION</b>The soothing liver and invigorating spleen recipes may regulate hepatocyte inflammatory injury of rats through TLR4/p38MAPK signaling pathway.</p>


Subject(s)
Animals , Female , Humans , Male , Rats , Drugs, Chinese Herbal , Hepatocytes , Metabolism , Lipopolysaccharides , Liver , Wounds and Injuries , Metabolism , Non-alcoholic Fatty Liver Disease , Drug Therapy , Genetics , Metabolism , Rats, Sprague-Dawley , Signal Transduction , Spleen , Metabolism , Toll-Like Receptor 4 , Genetics , Metabolism , p38 Mitogen-Activated Protein Kinases , Genetics , Metabolism
16.
National Journal of Andrology ; (12): 72-76, 2013.
Article in Chinese | WPRIM | ID: wpr-256923

ABSTRACT

<p><b>OBJECTIVE</b>To investigate sperm DNA integrity in male infertility patients with hepatitis B virus (HBV) infection.</p><p><b>METHODS</b>This study included 90 infertile men with HBV infection (group A), 82 infertile men without HBV infection (group B) and 70 normal fertile men (group C). We detected sperm DNA integrity among the subjects, including DNA fragmentation index (DFI) and high DNA stainability (HDS), by sperm chromatin structure assay (SCSA), and compared them among the three groups.</p><p><b>RESULTS</b>DFI was higher in group A ([28.17 +/- 13.06]%) than in B ([26.64 +/- 9.79]%) and C ([15.67 +/- 4.73]%), significantly higher in A and B than in C (P < 0.05) but with no significant difference between A and B (P > 0.05). HDS was higher in group A ([10.83 +/- 5.601]%) than in B ([9.04 +/- 3.48]%) and C ([8.04-2.25]%), with significant difference between A and C (P < 0.05).</p><p><b>CONCLUSION</b>Sperm DNA integrity of infertile males is significantly different from that of normal fertile men, and infertility with HBV infection further impairs sperm DNA, which is manifested by abnormal sperm nuclear maturity.</p>


Subject(s)
Adult , Humans , Male , Young Adult , Case-Control Studies , Chromatin , DNA , Genetics , DNA Damage , Hepatitis B , Pathology , Hepatitis B virus , Infertility, Male , Genetics , Virology , Sperm Count , Spermatozoa , Pathology
17.
Chinese Journal of Applied Clinical Pediatrics ; (24): 395-397, 2013.
Article in Chinese | WPRIM | ID: wpr-732981

ABSTRACT

Objective To compare the therapeutic effects of embryonic nature orifice transumbilical endoscopic surgery(ENOTES) with conventional laparoscopic(CL) surgery in Hirschsprung disease(HD).Methods A total of 111 children from Sep.2009 to Dec.2010 were included in the retrospective study.The patients were divided into 2groups:ENOTES group (66 cases) and CL group (45 cases).The general conditions of children were reviewed operative situation,postoperative complications and defecation 1 year after operation.Results In the left colectomy,there was no difference between 2 groups in age,weight,operative blood loss and postoperative stay (all P > 0.05),but ENOTES took up less time than CL(P < 0.05) ;in the subtotal colectomy,no significant difference existed in all series.All children underwent primary radical surgery,neither transfer to open surgery nor dead case.In ENOTES group,there were 9 cases suffering from enterocolitis postoperatively (13.6%),while 7 cases (15.6%) in CL group.Twenty-eight patients in ENOTES group and 12 in CL group were followed up for 1 year.Less abdominal distension postoperatively was noted in ENTOES group than CL group(P < 0.05),but no other difference in other available data.In addition,no obvious operative scar was noted on the abdomen in ENOTES group,what meant better cosmetic benefit than CL.Conclusion HD treated with ENOTES can gain the same outcomes with CL with better cosmetic effect.

18.
Chinese journal of integrative medicine ; (12): 127-131, 2013.
Article in English | WPRIM | ID: wpr-293299

ABSTRACT

<p><b>OBJECTIVE</b>To observe the therapeutic efficacy and the mechanism of Chinese medicine eye-patch with invigorating blood circulation and detoxification on asthenopia.</p><p><b>METHODS</b>A total of 180 rabbits were subjected to three tests, namely the skin microcirculation, the microvascular regeneration in the skin and the skin temperature change, with 60 rabbits for each test. The rabbits in each test were randomly and equally divided into three groups: the normal control group was treated with physiological saline on naked back once per day, the low dose group was treated with the eye-patch on naked back for 15 min once per day, while the high dose group was treated for three times per day. Forty Hartley guinea pigs were randomly and equally divided into four groups for the muscle tone test: the normal control group was treated with physiological saline on naked back, the model group was treated without any medication, the low dose group was treated with the eye-patch on naked back for 10 min once per day, while the high dose group was treated for three times per day. All treatments were continued for 14 days until the termination of the test. The microcirculatory blood flow was observed by using a video-microscopy system. The histological sections were used to detect the microvascular regeneration by observing the expression of factor VIII. The temperature changes on the skin surface were measured by using infrared thermometer, and the muscle tone was tested by the electromyography.</p><p><b>RESULTS</b>In compare with the normal or the model group, the improvement in the skin microcirculation and the blood vascular regeneration, and the decreasing in the muscle tone in low dose and high dose groups were statistically significant with confident level at P<0.05.</p><p><b>CONCLUSIONS</b>The eye-patch with invigorating blood circulation and detoxification has great enhancement in blood vascular regeneration and skin microcirculation, and great improvement in the indexes for muscle tone. The study explains certain therapeutic efficacy and mechanism of the eye-patch and shows that it could reduce the symptoms for patients with asthenopia.</p>


Subject(s)
Animals , Rabbits , Asthenopia , Drug Therapy , Dose-Response Relationship, Drug , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Guinea Pigs , Microcirculation , Protective Agents , Pharmacology , Therapeutic Uses , Regeneration , Regional Blood Flow , Skin , Skin Temperature
19.
Chinese Medical Journal ; (24): 1333-1336, 2013.
Article in English | WPRIM | ID: wpr-342181

ABSTRACT

<p><b>BACKGROUND</b>We previously reported that iodine-131((131)I)-labeled anti-pro-gastrin-releasing peptide (ProGRP(31-98)) monoclonal antibody D-D3 could selectively accumulate in the tumor sites of nude mice bearing small cell lung cancer (SCLC) xenografts. However, (131)I-D-D3 was cleared slowly from the body, and the best radioimmunoimaging time for SCLC was 72 - 96 hours after injection. The aims of this study were to radiolabel anti-ProGRP(31-98) D-D3 monoclonal antibody with technetium-99m ((99m)Tc) and to investigate the biodistribution of this antibody in healthy ICR mice.</p><p><b>METHODS</b>D-D3 was labeled with (99m)Tc via the 2-mercaptoethanol reduction method. (99m)Tc-D-D3 was purified by the gel column separation method. The labeling efficiency and radiochemical purity were measured by thin-layer chromatography. The immunological activity of (99m)Tc-D-D3 was determined with cell conjugation assays. (99m)Tc-D-D3 was injected into healthy ICR mice via a tail vein, and all the healthy ICR mice were sacrificed by cervical dislocation at a designated time. Then, the blood and major organs were removed and weighed, and counted in a gamma scintillation counter to determine the percentage of the injected dose per gram (%ID/g).</p><p><b>RESULTS</b>The labeling rate and the radiochemical purity of (99m)Tc-D-D3 were (73.87 ± 2.89)% and (94.13 ± 4.49)%, respectively. The immunobinding rates of (99m)Tc-D-D3 to the human small cell lung cancer NCI-H446 cell line and lung adenocarcinoma A549 cell line were (81.2 ± 2.37)% and (24.3 ± 1.46)%, respectively. The distribution data of normal ICR mice demonstrated that (99m)Tc-D-D3 was mainly distributed in the liver, kidney and lung, and less in the brain tissue and muscle.</p><p><b>CONCLUSIONS</b>(99m)Tc-D-D3 antibody not only had high radiochemical purity, but also had good stability both in vitro and in vivo, and maintained good immunological activity. (99m)Tc-D-D3 was metabolized mainly in the kidney and liver, and the blood radioactivity decreased rapidly. Thus, (99m)Tc-D-D3 is conducive to the radioimmunoimaging of SCLC.</p>


Subject(s)
Animals , Female , Male , Mice , Antibodies, Monoclonal , Chemistry , Allergy and Immunology , Metabolism , Mice, Inbred ICR , Peptide Fragments , Allergy and Immunology , Recombinant Proteins , Allergy and Immunology , Technetium , Chemistry
20.
Chinese Journal of Pediatrics ; (12): 527-530, 2013.
Article in Chinese | WPRIM | ID: wpr-275661

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the clinical characteristics, diagnosis and therapy of Keutel syndrome, and thereby to minimize the misdiagnosis.</p><p><b>METHOD</b>Data of a case of Keutel syndrome diagnosed at the Provincial Hospital Affiliated to Shandong University were analyzed and related literature were reviewed.</p><p><b>RESULT</b>An 8-month-26-day-old boy was presented with inspiratory and expiratory stridor and wheezing after movement on lung auscultation. His craniofacial appearance was characterized by midfacial hypoplasia with a broad depressed nasal bridge. The nose was small and flat. A grade 2-3/6 systolic murmur was heard between the second and third ribs at left edge of the sternum. The end phalanges of his fingers were thickened. Chest radiograph showed tracheobronchial cartilage calcification and tracheobronchial stenosis. Echocardiographic examination revealed the right pulmonary stenosis. With endoscopic surgery, antiobstructive and antibiotic therapy clinical symptoms were improved. Three weeks later he died of lung reinfection after he was discharged from our hospital. English literature search with "Keutel syndrome" as the key word at "PubMed" showed 22 articles covering 26 patients, and the clinical symptoms were hearing loss (91%), persistent respiratory symptoms (68%), recurrent otitis media/sinusitis (67%), growth development delay (52%) in turn, and signs were brachytelephalangism (100%), low nasal bridge (95%), midfacial hypoplasia (93%), cardiac murmur (69%), and auxiliary examinations showed abnormal cartilage calcification (100%), pulmonary arterial stenosis (72%), tracheobronchial stenosis (50%).</p><p><b>CONCLUSION</b>The diagnosis of Keutel syndrome should be considered in patients with brachytelephalangism, abnormal cartilage calcification, peripheral pulmonary stenosis, and midfacial hypoplasia. Tracheal stenosis was main clinical manifestation in part of patients.</p>


Subject(s)
Humans , Infant , Male , Abnormalities, Multiple , Diagnosis , Diagnostic Imaging , Therapeutics , Bone and Bones , Diagnostic Imaging , Calcinosis , Diagnosis , Diagnostic Imaging , Therapeutics , Cartilage , Diagnostic Imaging , Cartilage Diseases , Diagnosis , Diagnostic Imaging , Therapeutics , Diagnosis, Differential , Hand Deformities, Congenital , Diagnosis , Diagnostic Imaging , Therapeutics , Pulmonary Valve Stenosis , Diagnosis , Diagnostic Imaging , Therapeutics , Radiography, Thoracic , Retrospective Studies , Tomography, X-Ray Computed , Tracheal Stenosis , Diagnosis , Diagnostic Imaging
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