Objective To investigate the variations of the expression and activity of phosphatidyl inositol-3 kinase(PI-3K)in the adipose tissue of pregnant women with gestational diabetes mellitus(GDM)and to explore its relationship with insulin resistance(IR)in these women.Methods The expression of PI-3K p85aprotein and mRNA in adipose tissue were detected by Western blot and Semi quantitative reverse transcription polymerase chain reaction(RT-PCR)in 20 GDM women(GDM group)and 20 normal pregnant women(normal group).The activity of PI-3K was determined by ELISA and IR index was calculated using homeostasis model assessment(HOMA)according to the results of fasting insulin(FINS)and fasting plasma glucose(FPG),which measured by oxidase assay and immunoradioassay,respectively.Results The expressions of PI-3K p85αmRNA and PI-3K p85a protein were markedly increased in the adipose tissue of GDM group than in the normal group(mRNA:0.83±0.03 vs 0.53±0.07,P＜0.01;protein:0.93±0.04 vs 0.71±0.06,P＜0.01).However,the PI-3K activity in the GDM groups was significantly down-regulated compared with the normal group(1.7±0.6 vs 5.2±0.5,P＜0.01).The levels of FPG and FINS (HOMA-IR)in the GDM group were all significantly higher than the normal group[FPG:(5.8±0.2)mmol/L vs(4.7±0.3)mmol/L;FINS:(14.8±0.2)mmol/L vs(11.2±0.3)mmol/L;HOMA-IR:1.3±0.4 vs 0.9±0.3,a11 P＜0.01]. Conclusions The decreased activity of PI-3K in the adipose tissue may be one of the molecular mechanisms for IR in GDM.

Objective To investigate the expression and tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1) and in the adipose tissue of patients with gestational diabetes mellitus (GDM), and to explore molecular mechanisms of insulin resistance in GDM. Methods The serum and adipose tissue were sampled from patients with GDM (GDM group, n = 20) and normal pregnant women (control group, n = 20). Fasting plasma glucose was measured by glucose oxidase assay. The expressions of IRS-1 protein and mRNA were determined by Western blot and semi-quantitative RT-PCR. The tyrosine phosphorylation of IRS-1 was measured by immunoprecipitation. Results Compared with control group, in GDM group, the expression of IRS-1 mRNA was markedly decreased (0.61 ±0.06 vs 1.12 ± 0.17, P < 0.01), the expression of IRS-1 protein was significantly decreased (0.57 ±0.08 vs O. 83 ±0.07, P <0.01) and tyrosine phosphorylation was significantly reduced (0.23 ± O. 06 vs O. 62 ±0.04, P < 0.01) in the adipose tissue. Conclusion The decline of protein expression and tyrosine phosphorylation of IRS-1 in the adipose tissue of gestational diabetes appears to be one of the moleculemechanisms of insulin resistance in patients with GDM.

Aim To investigate the relationship be-tween the cardioprotection of apigenin ( Api ) from an-oxia/reoxygenation ( A/R) injury and Bcl-2 pathway. Methods H9 c2 cardiomyocytes were cultured and di-vided into normal control group, A/R group, Api pre-treatment group ( Api ) , Api + Bcl-2 inhibitor group ( Api + ABT-737 ) . Expression of Bcl-2 was deter-mined by Western blot,and cell viability was measured by MTT method. LDH, SOD, GSH-Px, MDA activity were determined by chromometry. ROS generation, mi-tochondrial membrane potential and apoptosis were de-termined by flow cytometry. Results 25h after apige-nin precondition,the expression of Bcl-2 was upregulat-ed in cardiomyocytes ( P <0. 01 ) . In the group pre-treated with 40 μmol · L-1 apigenin before A/R, the activity of LDH in culture medium decreased; the ac-tivity of intracellular SOD, GSH-Px increased; the content of MDA and ROS generation decreased; cell viability increased; mitochondrial membrane potential could be more stable and cell apoptosis decreased ( P<0. 01 ) . However, all these protective effects were attenuated significantly in the group pretreated with apigenin and Bcl-2 inhibitor ABT-737 . Conclusion The effect of apigenin against A/R injury in cardiomyo-cytes involves Bcl-2 pathway, and at least partly de-pends on its effect on upregulating the expression of Bcl-2 .

AIM: To observe the effects of Qi Jing Mingmu decoction combined with artificial tears on the clinical results and cell aging of conjunctivochalasis.

METHODS: Forty cases(80 eyes)of grade II-Ⅳ CCH with liver-kidney Yin deficiency were randomly divided into two groups: combined treatment group and artificial tears group, which were treated with Qi Jing Mingmu decoction combined with artificial tears and simple artificial tears respectively. The international ocular surface disease index(OSDI), tear break-up time(BUT)and Schirmer Ⅰ test(SⅠt)were observed and the clinical effects were compared after 3mo treatment. For CCH patients with grade III or above, followed up for 3mo or more and willing to operate, the loose conjunctival tissue was removed and the cell aging related β-gal staining was performed on frozen sections. The results were statistically analyzed.

RESULTS: The OSDI score(14.53±2.68), BUT 9.25±3.02s and SⅠt(8.95±3.57mm/5min)of combined treatment group were significantly better than those of artificial tears group after 3mo treatment(all P<0.05). After drug treatment, 7 cases(7 eyes)in artificial tears group and 4 cases(4 eyes)in combined treatment group of CCH patients were treated by operation. The positive rate of aging cells in combined treatment group was significantly lower than that in artificial tears group(16.00±7.84 vs 39.00±14.09, P=0.013).

CONCLUSION: Qi Jing Mingmu decoction combined with artificial tears to treat CCH is more effective than simple artificial tears in relieving ocular symptoms, improving tear film and promoting tear secretion. Combined treatment can also reduce the cell aging in CCH, which can be used as a safe and effective treatment method in addition to surgical operation.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Neoplasm Invasiveness ; Neoplasm Metastasis ; PTEN Phosphohydrolase ; biosynthesis ; genetics ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

The primary object of this fundamental research was to survey the synergistic cardiovascular effects of iptakalim, a novel ATP-sensitive potassium channel (K(ATP)) opener, and clinical first-line antihypertensive drugs, such as calcium antagonists, thiazide diuretics and β receptor blockers by a 2 x 2 factorial-design experiment. It would provide a theoretical basis for the development of new combined antihypertensive therapy program after iptakalim is applied to the clinic. Amlodipine besylate, hydrochlorothiazide and propranolol were chosen as clinical first-line antihypertensive drugs. Blood pressure, heart rate (HR) and cardiac functions were observed in anesthetized normal rats by an eight-channel physiological recorder. The results showed that iptakalim monotherapy in a low dose could produce significant antihypertensive effect. There was no interaction between iptakalim and amlodipine on the maximal changes of systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial blood pressure (MABP), the left ventricular systolic pressure (LVSP), and the left ventricular end-diastolic pressure (LVEDP) (P > 0.05). However, the effects of combination iptakalim/amlodipine on the maximal changes of SBP, DBP, MABP, LVSP and LVEDP were more obvious than those of iptakalim or amlodipine monotherapy. And there was strong positive interaction between iptakalim and amlodipine on the maximal changes of HR (P>0.05). According to the maximal changes of DBP, MABP, LVSP and LVEDP (P < 0.05) of combination iptakalim with hydrochlorothiazide, there was strong positive interaction between them. But there was no interaction between iptakalim and hydrochlorothiazide on the maximal drop of SBP and HR (P > 0.05). According to the maximal drops of DBP, MABP of combination iptakalim with propranolol, there was strong positive interaction between them (P < 0.05). But there was no interaction between iptakalim and propranolol on the maximal changes of SBP, LVSP, LVEDP and HR (P > 0.05). In conclusion, it was the first time to study the effects of amlodipine, hydrochlorothiazide or propranolol, which had different mechanisms of action from iptakalim, on cardiovascular effects of iptakalim in anesthetized normal rats. This study proved that the combination of iptakalim with hydrochlorothiazide or propranolol respectively had significant synergism on lowering blood pressure, while the combination of iptakalim/amlodipine had additive action on lowering blood pressure. Meanwhile the antihypertensive effect was explicit, stable and long-lasting. Iptakalim thus appears suitable for the clinical treatment of hypertensive people who need two or more kinds of antihypertensive agents.
Amlodipine ; pharmacology ; Animals ; Antihypertensive Agents ; pharmacology ; Blood Pressure ; drug effects ; Drug Synergism ; Heart Rate ; Hydrochlorothiazide ; pharmacology ; Hypertension ; Propranolol ; pharmacology ; Propylamines ; pharmacology ; Rats

Background Researches showed that the incidence rate of cataract is high in the nickel mining area. Nickel sulphate can apparently inhibit the metabolism and proliferation of human lens epithelium cells. But the study on the injury mechanism of nickel on lens is still seldom. Objective Present study was to investigate the effect of nickel sulphate on the lens of SD rats. Methods Forty-five SPF SD rats aged from 7 to 14 days were grouped randomly into subcutaneous injection group, intraperitoneal injection group and blank group. Nickel sulphate of 2 g/L ( 10 mg/kg) was subcutaneously or intraperitonealy injected for 45 days. The opacity of rat lens was examined under the slit lamp at two-week interval and scored based on the criteria of LOCS II and LOCS III. The rats were sacrificed in 45 days after experiment and the lens were obtained for the pathological examination. Result The mean score of the anterior subcapsule opacity of rat lens was obviously higher in subcutaneous injection group compared with blank control group with a significant difference between them (t= 14. 311, P < 0. 05 ) , but no significant difference in the anterior subcapsule opacity between intraperitoneal injection group and blank control group (t = 4. 355 , P>0. 05 ). The score of posterior subcapsule opacity of lens were evidently higher in both subcutaneous injection group and intraperitoneal injection group than the blank control group (t = 9. 316,P = 0. 004;t = 7. 464, P = 0. 009) ,so was the mean score of the anterior +posterior subcapsule opacities(t = 23. 387,P=0. 000;t= 10. 533,P = 0. 002) and the total score of rat lens opacity ( t = 12. 358 , P = 0. 001; t = 10. 188 , P = 0. 003 ) . No significant differences were found in cortex opacity score and nuclear opacity score among three groups ( P > 0.05 ). Histopathology examination revealed that the degeneration of lens collagen protein was more serious in subcutaneous injection group and intraperitoneal injection group than the blank control group,and the injury degree of lens collagen protein was more dominant in subcutaneous injection group. Conclusion System administration of nickel sulphate induced the injury of anterior and posterior subcapsule of lens in SD rat.

Objective To establish a homothermal and fast detecting method on pathogenic bacteria by combining recombinase-aid amplification (RAA) with molecular beacon.Methods The establishment of the methodology.Staphylococcus aureus specific primers were designed from the relative region of the staphylococcal protein A (SPA).Asymmetry amplification was optimized by adjusting the primer concentration ratios.The results of amplification and hybridization were visualized and analyzed by agarose gel electrophoresis and fluorescence detection.The sensitivity was identified by detecting dilute positive plasmids.And the specificity was determined using RAA method by detecting 72 pathogenic bacteria,including Staphylococcus aureus and other Staphylococcus spp.from the Department of Clinical Laboratory of Daping Hospital in December 2016.Besides,the Kappa analysis and the clinical diagnosis efficiency were investigated by analyzing 39 extra strains in the laboratory in December 2016.Results When the concentration ratio of restrictive and non-restrictive primer was 1:20,the yield efficiency of single-stranded DNA (ssDNA) reached the peak.And as for the hybridization efficiency,the asymmetry amplification was higher than symmetry amplification.Twenty copies/μl was proposed as the limits of detection by testing dilute plasmids.And the RAA hybridization method could distinguish Staphylococcus aureus with other Staphylococcus spp.Comparing with traditional detection methods with a Kappa index of 0.860,this method shows a good consistency.By analyzing the 111 bacteria,the sensitivity of the method is 92.5％ (37/40),the specificity is 97.2％ (69/71),the positive predictive value is 94.9％ (37/39),the negative predictive value is 95.8％ (69/72),the positive likelihood radio is 33.04,the negative likelihood radio is 0.077,the Youden index is 0.897 and the Kappa index is 0.902.Conclusion Through the combination of asymmetry recombinase-aid amplification optimization and molecular beacon probe,a new method of detecting bacteria DNA with RAA hybridization technique is established,providing the foundation for its clinical application.

OBJECTIVETo explore the effects of integrative medicine (IM) rehabilitation protocolon motor function, activity of daily living, and quality of life (QOL) in hemiplegia patients after stroke.

METHODSTotally 120 patients with post-stroke hemiplegia were allocated to four groups using sealed envalope drawing, i.e., the rehabilitation group, the Chinese medical treatment group, the acupuncture group, and the comprehensive rehabilitation group, 30 cases in each group. Based on routine rehabilitative training, patients in the Chinese medical treatment group, the acupuncture group, and the compre-hensive rehabilitation group received standardized treatment based on syndrome typing, Shi's Consciousness-Restoring Resuscitation acupuncture, Chinese herbs + acupuncture comprehensive rehabilitatino protocol, respectively. The treatmet cycle consisted of 4 weeks with 24-week follow-ups. Fugl-Meyer motor assessment (FMA), Modified Barthel Index (MBI), and Stroke-Specific Quality of Life Scale(SS-QQL), and safety assessment were taken as main effect indices before treatment, at week 4 of treatment, at week 12 and 24 of follow-ups, respectively.

RESULTSThere was no statistical difference in FMA score, MBI score, SS-QOL score among the four groups before treatment (P > 0.05). These scores were significantly improved in the four groups at week 4 of treatment, week 12 and 24 of follow-ups, respectively (P < 0.05). Besides, FMA score and SS-QOL score were significantly improved in the comprehensive rehabilitation group at each corresponding time point, as compared with other treatment groups (P < 0.05).

CONCLUSIONSThe comprehensive protocol could significantly improve motor function, activity of daily living in hemiplegia patients after stroke, and further improve their QOL. Its effect was better than other single treatment.

Activities of Daily Living ; Acupuncture Therapy ; Hemiplegia ; rehabilitation ; Humans ; Integrative Medicine ; methods ; Medicine, Chinese Traditional ; Motor Skills ; Quality of Life ; Stroke Rehabilitation ; Treatment Outcome

OBJECTIVETo investigate the possible correlation between atherosclerosis and chronic periodontitis by establishing an animal model of chronic periodontitis and atherosclerosis in Wistar rat.

METHODSSixty male Wistar rats were divided into four groups: A (control group), B (chronic periodontitis group), C (atherosclerosis group), D (chronic periodontitis accompany with atherosclerosis group). Every group was accepted the corresponding treatment. Animals were sacrificed after 12 weeks. The periodontal index, levels of serum total cholesterol (TC) and low-density lipoprotein (LDL), the concentration of TNF-alpha and matrix metalloproteinase (MMP-3) were examined. The severity of chronic periodontitis and atherosclerosis was quantified by histopathology. The date were statistically analyzed.

RESULTSThrough detection of periodontal tissue of experimental teeth, serum and histopathology, animal models were successful. Histopathologic observation revealed:obvious inflammation of periodontal tissue was observed in group B and D. Attachment loss level in group B [(137.86 +/- 28.39) microm] and D [(162.36 +/- 22.69) microm] was higher than that in group A [(4.26 +/- 1.07) microm] and C [(68.07 +/- 18.25) microm] (P < 0.05), and that in group C was higher than group A (P < 0.05). Atherosclerotic lesions of abdominal aorta were formed in group C and D. The level of TC, LDL in group C and D was higher than that in group A and B (P < 0.05), and that in group D was higher than group C (P < 0.05). Animals in group B and D showed higher level of TNF-alpha, MMP-3 in serum than that in group A and C (P < 0.05). There was no correlation between the level of MMP-3 and TC (P = 0.971) or LDL (P = 0.604).

CONCLUSIONSChronic periodontitis may be a risk factor and contribute to the pathogenesis of atherosclerosis. MMP-3 may be an independent risk factor of atherosclerosis exclude TC and LDL.

Animals ; Aorta, Abdominal ; Aortic Diseases ; etiology ; pathology ; Atherosclerosis ; etiology ; pathology ; Cholesterol ; blood ; Chronic Periodontitis ; complications ; pathology ; Disease Models, Animal ; Drugs, Chinese Herbal ; Lipoproteins, LDL ; blood ; Male ; Matrix Metalloproteinase 3 ; blood ; Periodontal Index ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; blood