Objective:To investigate the expression of serum caspase-cleaved cytokeratin 18(CCCK-18) in patients with cerebral ischemic stroke and its diagnostic value.Methods:One hundred and six patients with cerebral ischemic stroke who were diagnosed and treated in Affiliated Dongfeng Hospital from October 2018 to October 2019 were selected as the study group. Ninety patients who underwent digital subtraction angiography (DSA) during the same period and showed no other abnormalities inside or outside the skull were selected as control group. The baseline data of gender, age, drinking history, smoking history, hypertension history, diabetes history, coronary heart disease, and other subjects in the two groups had no significant differences ( P>0.05). Cubital venous blood of 5 ml from two groups of subjects were collected, and the level of serum CCCK-18 was detected by enzyme-linked immunosorbent assay. The levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) were detected by enzymatic method. The receiver operating characteristic curve (ROC curve) was used to analyze the diagnostic efficacy of serum CCCK-18 in patients with ischemic stroke, and the relationship between serum CCCK-18 and TC, TG, LDL-C, HDL-C were analyzed by Pearson test. Results:The levels of serum CCCK-18, TC, TG, and LDL-C in the study group were significantly higher than those in the control group: (158.10 ± 50.89) U/L vs. (85.57 ± 35.25) U/L, (4.26 ± 0.92) mmol/L vs. (3.92 ± 0.80) mmol/L, (2.34 ± 0.53) mmol/L vs. (1.83 ± 0.47) mmol/L, (3.12 ± 0.73) mmol/L vs. (2.61 ± 0.67) mmol/L, and HDL-C level was lower than that in the control group: (1.20 ± 0.24) mmol/L vs. (1.32 ± 0.28) mmol/L, and there were significant differences ( P<0.05). Logistic regression analysis showed that CCCK-18, TC, TG, LDL-C, and HDL-C were independent risk factors for patients with ischemic stroke ( P<0.05). The area under the curve(AUC) of serum CCCK-18 to distinguish ischemic stroke from the control group was 0.878, with a sensitivity of 84.91% and a specificity of 78.89%. The AUC of serum CCCK-18 to identify patients with mild ischemic stroke was 0.763, with a sensitivity of 70.37% and a specificity of 78.89%. Correlation analysis showed that serum CCCK-18 was positively correlated with TC, TG, and LDL-C in patients with ischemic stroke ( r = 0.711, 0.722, 0.705), and negatively correlated with HDL-C ( r = - 0.714), and there were significant differences ( P<0.01). Conclusions:Serum CCCK-18 levels are significantly increased in patients with cerebral ischemic stroke, which can be used as a biomarker for diagnosis and judgment of disease severity.

Objective In order to investigate the roles of hIGF\|1 in treatment of diabetes mellitus and diabetic syndromes,the gene of human insulin like growth factor type Ⅰ(IGF\|1) was cloned and constructed into eukaryotic expression vector,then the expression and activity were determined. Methods Total cellular RNA of human fetal liver was abstracted and the RT\|PCR amplification of the cDNA fragment was performed.The fragment was cloned into pUCM\|T vector and sequenced.The eukaryiotic expression vector was recombined and transfected into fibroblast cell line,COS\|7.The expression of hIGF\|1 was examined by in situ hybridization and immunohistochemistry.The effect of hIGF\|1 on cultured islet cells was observed by glucose\|stimulated insulin release assay. Results The cDNA fragment of 710bp with additional Kozak sequence was amplified by RT\|PCR.Eukaryiotic expression vector pCI\|neo/hIGF\|1 was constructed and IGF\|1 gene expressed in COS\|7.The biological activity of hIGF\|1 was proved by increasing inslin secretion from islet cells.Conclusion\ The newly constructed vector,pCI\|neo/hIGF\|1 could be transfected into COS7 cells and its expressed product showed to have the biology activity of hIGF\|1.\;[

10.3969/j.issn.2095-4344.2013.23.026

Aim To investigate the interactions be-tween the neuroinflammation caused by lipopolysaccha-ride(LPS) and brain CYP2E1.Methods The human cholinergic neuroblastoma cell line IMR-32 was treated with LPS ( 0.1 mg · L-1 , 1.0 mg · L-1 ) , and the LDH and SOD activities were determined after 24 h in-cubation .In order to determine the roles of MAPK sig-naling pathway in the regulation of CYP 2E1 by LPS, the IMR-32 cells were treated with p38 pathway inhibi-tor SB203580 or ERK pathway inhibitor U 0126 for 45 min before the incubation with LPS .The human do-paminergic neuroblastoma cell line SH-SY5Y with CYP2 E1 over-expression was established . The LDH and SOD activities were determined in SH-SY5 Y cells over-expressed CYP2 E1 and control cells treated with LPS(0.1 mg· L-1 , 1.0 mg· L-1 ) for 24 h.Results
The levels of LDH in IMR-32 cells treated with high-dose LPS were increased by 1.38-fold ( P <0.01 ) compared with the control group , and the levels of SOD reduced by 15.0%( P <0.01 ) .Compared with the control, CYP2E1 mRNA and protein levels in IMR-32 cells treated with high-dose LPS were increased by 1.25-fold(P<0.01) and 1.19-fold(P<0.05).The up-regulation of CYP2E1 by LPS could be attenuated by SB203580 and U0126 pretreatment.Compared with the control cells, the CYP2E1 over-expression in-creased LDH levels by 1.28-fold ( P<0.01 ) and de-creased SOD levels by 3.53-fold ( P<0.01 ) after the low-dose of LPS treatment .The CYP2E1 over-expres-sion increased LDH levels by 1.54-fold ( P <0.01 ) and decreased SOD levels by 2.17-fold( P<0.01) af-ter the high-dose of LPS treatment , compared with the control cells.Conclusions LPS can induce CYP2E1 mRNA and protein levels , and the p38 and ERK sig-naling pathway may be involved in the regulation .The elevated CYP2 E1 levels aggravate the damage to neuro-nal cells caused by LPS .Brain CYP2E1 may be an im-portant contributing factor to the pathological process of neuroinflammatory injury .

Objective To compare the infectivity between laboratory adapted human inununodefi- ciency virus(HIV-1) and primary HIV-1 isolates for different mucosal epithelial cell lines. Methods Mu-cosal epithelial cells Caco-2, T-84, HeLa and lymphocyte MT-4 were infected with laboratory adapted HIV-1 SF33 and 2 primary HIV-1 isolates (02010561, 02010141). Culture supernatant and cells were collected respectively on 3-4 days interval after virus inoculation. The former was tested for HIV-1 antigen P24 level and viral load, and the latter was tested for total viral DNA and integrated viral DNA. Results All 3 virus strains could infect MT-4 cells and integrate into their genome. Only HIV-1 SF33 could infect Caco-2 cells but could not integrate into their genomic DNA. Both HIV-1 SF33 and 02010561 infected HeLa cells but only integration of HIV-1 SF33 was detected. All the 3 HIV-1 strains infected T-84 cells but only the integra-tion of HIV-1 SF33 and 02010141 was observed. Conclusion Although laboratory adapted and primary HIV-1 strains are able to infect human mucosal epithelial cell lines, transient or productive infection estab-lished in different mucosal epithelial cells is dependent on the character of cells and virus strains.

Purpose To detect the expressions of stress induced phosphoprotein 1 (STIP1) and estrogen receptor-α(ER-α) in papil-lary thyroid carcinoma and to analyse the relationship between STIP1 and ER-α. Methods 54 cases of paraffin-embedded tissues of papillary thyroid carcinoma, 18 cases of papillary thyroid carcinoma with lymph node metastasis, 15 cases of Hashimoto’ s thyroiditis, 10 cases of adjacent normal thyroid tissue were collected. The expressions of STIP1 and ER-αwere detected by immunohistochemistry, and the relationship between the expressions and clinicopathological features of papillary thyroid carcinoma was analyzed. Results The expression of STIP1 and ER-α in papillary thyroid cancer group ( 55. 6% and 44. 4%) were higher than that of normal thyroid group (10% and 0) and Hashimoto’s thyroiditis group (8. 3% and 0, all P<0. 05). STIP1 expressions was related to lymph node metastasis ( P<0. 05 ) , while ER-α expression was related to gender, TG-Ab and the merger of nodular goiter, but not related to lymph node metastasis (P>0. 05). The expressions of STIP1 and ER-α in papillary thyroid carcinoma were not related to patients’ age , tumor location, number of tumors, tumer size, invasion of capsule, the concomitant Hashimoto’ s thyroiditis and TPO-Ab ( all P>0. 05). And the expressions of STIP1 was not related to gender, TG-Ab and the merger of nodular goiter (all P>0. 05). A positive correlation was found between the expressions of STIP1 and ER-αin thyroid papillary carcinoma (P<0. 05). Conclusion STIP1 and ER-α in papillary thyroid carcinoma may be related with lymph node metastasis.

OBJECTIVETo investigate the effect of oligochitosan in promoting intestinal absorption of protoberberine alkaloids in extracts from Corydalis saxicola total alkaloids.

METHODThe in vitro single-pass intestinal perfusion model in rats was established to study the changes in absorption kinetic parameters of dehydrocavidine, berberine hydrochloride and palmatine chloride in C. saxicola total alkaloids after the addition of different concentrations oligochitosan and evaluate the effect of oligochitosan in promoting intestinal absorption of the drugs.

RESULTThe concentration of oligochitosan had different effects on the absorption rate constant (Ka) and apparent permeability coefficient (Peff) of the three active component in rat intestines. Ka and Peff in 0.5% oligochitosan group significantly increased, indicating a stronger effect in promoting the absorption.

CONCLUSIONOligochitosan has a certain effect in promoting the intestinal absorptions of protoberberine alkaloids in C. saxicola total alkaloids.

Animals ; Berberine Alkaloids ; administration & dosage ; pharmacokinetics ; Chitin ; administration & dosage ; analogs & derivatives ; Corydalis ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo compare the efficacy difference in the treatment of functional dyspepsia between acupuncture at the acupoints selected by pattern/syndrome differentiation and domperidone.

METHODSSeventy cases were randomized into an acupuncture group (35 cases) and a western medication group (35 cases). In the acupuncture group, Zusanli (ST 36) and Neiguan (PC 6) were selected. Taichong (LR 3) and Neiting (ST 44) were added for excess syndrome while Gongsun (SP 4) and Yinlingquan (SP 9) were added for deficiency syndrome. A pair of electrodes was attached to one acupoint and an assistant point (2 mm next to the acupoint centripetally) and stimulated with disperse-dense wave at 2 Hz/100 Hz, once a day. In the western medication group, domperidone was prescribed for oral administration, 10 mg each time, three times a day. In the two groups, the treatment of 5 days made one session and 4 sessions were required totally. Nepean dyspepsia index (NDI) was compared after treatment, 1, 2, 3, 4 and 5 months after treatment between the two groups respectively.

RESULTSThe score of symptom and score of life quality in NDI after treatment and at each follow-up time point were improved obviously in the acupuncture group as compared with those before treatment (all P < 0.01). In the western mediation group, the score of symptom and the score of life quality in NDI after treatment and in follow-up of 1, 2 and 3 months were improved obviously as compared with those before treatment (all P < 0.01), but the differences were not significant in follow-up of 4 and 5 months (both P > 0.05). Compared with the western medication group, the symptom score of NDI was reduced obviously after treatment and in each time point of follow-up in the acupuncture group (P < 0.05, P < 0.01), and the score of life quality was increased obviously (P < 0.05, P < 0.01).

CONCLUSIONAcupuncture at the acupoints selected by pattern/syndrome differentiation and domperidone are effective in the treatment of functional dyspepsia. Domperidone is unsatisfactory in the long-term effect, but acupuncture achieves the positive short-term and long-term effects on functional dyspepsia.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Dyspepsia ; diagnosis ; psychology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Quality of Life ; Time ; Treatment Outcome ; Young Adult

The poor prognosis of hepatocellular carcinoma (HCC) is strongly associated with invasion and metastasis.Recently,the epithelial-mesenchymal transition (EMT) has been confirmed to be the cytological foundation of invasion and metastasis.Yet,the molecular mechanism of inducing and maintaining EMT has not been expounded completely.However,it has been demonstrated that transforming growth factor-β (TGF-β),phosphatidyl inositol 3-kinase/protein kinase B (PI3K/AKT),nuclear factor-κB (NF-κB),Wnt/β-catenin signaling pathways play pivotal roles in the initiation and development of EMT.These signaling pathways can affect the prognosis of HCC by regulating EMT.From a drug development perspective,these signal pathways are potential and attractive targets for HCC treatment.

Objective To observe the expression of synaptotagmin I(syt I)protein in the prefrontal cortex of adult-onset hypothyroidism rats and the effects of replicated therapy in different doses of thyroid hormone on the syt I protein.Methods All 44 aduh male Sprague-Dawley rats were divided into 4 groups randomly according to their body mass:hypothyroidism group,routine dosage thyroxine treatment group,high dosage thyroxine treatment group and control group.The adult male Sprague-Dawley rats were replicated to the adult-onset hypothyroidism and treatment models with propyhhiouracil(PTU).The levels of serum T3,T4 were assayed by the radioimmunoassay method and the level of the syt I protein in the molecular layer,external granular layer,external pyramidal layer,internal granular layer and internal pyramidal layer in prefrontal cortex was analyzed by immunohistochemistry.Results In the hypothyroidism group,the levels of serum T3 and T4[(0.34±0.04),(43.01±2.95)nmol/L]were significantly lower than those in the control group[(0.65±0.15), (55.20±3.56)nmol/L, F value: 6.026,5.940,4.503,P<0.05 or <0.01 ], the levels of the syt I protein in the molecular layer(0.018±0.010), external granular layer (0.020±0.007), external pyramidal layer(0.013±0.008), internal granular layer(0.011±0.005), internal pyramidal layer(0.024±0.013) of prefrontal lobe were significantly lower compared to the control group[(0.028±0.010,0.031 ± 0.010,0.028 ± 0.010,0.022 ± 0.008,0.038 ± 0.013), F value: 5.697,8.965,14.668,13.597,6.807,P<0.05 or <0.01 ]. In the routine dosage of the thyroxine treatment group, the levels of serum T3,T4 [(0.63 ±0.05), (55.04 ± 3.77)nmol/L] were not significantly different compared to the control group(F value: 3.162,0.367,all P>0.05), and the level of the syt I protein in the molecular layer, external granular layer, external pyramidal layer, internal granular layer and internal pyramidal layer in prefrontal cortex showed a significant improvement of the syt I protein(0.027 ± 0.013,0.025 ± 0.009,0.022 ± 0.008,0.020 ± 0.010,0.033 ± 0.010), which were similar to that of the control group(F value: 0.094,2.208,2.467,0.350,0.693, all P>0.05). In the high dosage thyroxine thyroid hormone treatment group, the levels of serum T3 and T4[ (1.11 ± 0.10), (96.68 ± 6.42)nmoL/L] were higher than the control group(F value: 6.291,12.031, all P<0.01), the expression of the syt I protein(0.028 ± 0.008,0.031 ±0.011,0.026 ± 0.012,0.023 ± 0.011,0.038 ± 0.010) were not significantly different compare to the control group (F value: 0.001,0.019,0.111,0.061,0.001, all P>0.05). Conclusions The expression of the syt I protein in the prefrontal cortex of adult-onset hypothyroidism can be decreased, which can be reversed by routine dosage of thyroxine treatment.