Investigating the mechanism of interactive regulation of B-cell lymphoma-2/Beclin 1 through electroacupuncture intervention during reperfusion in myocardial ischemia-reperfusion injury in a rat model.

To observe the regulation of B-cell lymphoma-2 (Bcl-2)/Beclin 1 interaction through electroacupuncture (EA) intervention during reperfusion and to investigate the EA mechanism of apoptosis-autophagy interactive regulation against myocardial ischemia-reperfusion injury (MIRI). A total of 48 adult Sprague Dawley (SD) rats were randomly divided into the sham-operated group (group Sham), the model group (group Model), the EA group (group EA), and the JNK inhibitor (SP600125) group (group JNK), with 12 rats in each group. Biospecimens were collected randomly from six rats in each group four hours after reperfusion. Evans Blue and triphenyl tetrazolium chloride double-staining were applied to observe each group's myocardial damage area and risk area. We collected 4 ml of blood by abdominal aortic method to detect serum troponin cTnI level by enzyme-linked immunosorbent assay (ELISA). For the remaining six in each group, a part of myocardial tissue below the ligation line was stored in 4% paraformaldehyde for immunohistochemistry and TUNEL staining; the other amount of myocardial tissue was detected by Western blotting to determine the expression levels of Bcl-2, Beclin1, and the phosphorylation levels of Thr69, Ser70, and Ser87 in Bcl-2. In results: electroacupuncture (EA) intervention during reperfusion significantly reduced the myocardial infarction area, cTnI level, and myocardial apoptosis, upregulated Bcl-2 expression, downregulated Beclin 1 expression and inhibited phosphorylation levels of Thr69, Ser70, and Ser87 in Bcl-2. We concluded that EA effectively inhibited apoptosis by upregulating Bcl-2 expression and inhibiting the phosphorylation of Thr69, Ser70, and Ser87 in Bcl-2. This reduced the separation of Bcl-2 and Beclin 1, restrains excessive autophagy, alleviates MIRI, and has a protective effect on myocardial tissue.

Folic acid supplementation and dietary folate intake, and risk of preeclampsia.

BACKGROUND/OBJECTIVES: Folic acid supplementation has been suggested to reduce the risk of preeclampsia. However, results from few epidemiologic studies have been inconclusive. We investigated the hypothesis that folic acid supplementation and dietary folate intake before conception and during pregnancy reduce the risk of preeclampsia. SUBJECTS/METHODS: A birth cohort study was conducted in 2010-2012 at the Gansu Provincial Maternity & Child Care Hospital in Lanzhou, China. A total of 10,041 pregnant women without chronic hypertension or gestational hypertension were enrolled. RESULTS: Compared with nonusers, folic acid supplement users had a reduced risk of preeclampsia (OR=0.61, 95% CI: 0.43-0.87). A significant dose-response of duration of use was observed among women who used folic acid supplemention during pregnancy only (P-trend=0.007). The reduced risk associated with folic acid supplement was similar for mild or severe preeclampsia and for early- or late-onset preeclampsia, although the statistical significant associations were only observed for mild (OR=0.50, 95% CI: 0.30-0.81) and late-onset (OR=0.60, 95% CI: 0.42-0.86) preeclampsia. The reduced risk associated with dietary folate intake during pregnancy was only seen for severe preeclampsia (OR=0.52, 95% CI: 0.31-0.87, for the highest quartile of dietary folate intake compared with the lowest). CONCLUSIONS: Our study results suggest that folic acid supplementation and higher dietary folate intake during pregnancy reduce the risk of preeclampsia. Future studies are needed to confirm the associations.

Effect of berberine on the pharmacokinetics of substrates of CYP3A and P-gp.

The in vivo effects of berberine (BBR), the widely used bioactive herbal ingredient from many traditional Chinese medicinal herbs, on the pharmacokinetics of carbamazepine (CBZ, a substrate of CYP3A) and its metabolite carbamazepine 10,11-epoxide (ECBZ), digoxin (DIG, a substrate of P-gp) and cyclosporine A (CsA, a dual substrate of CYP3A and P-gp) were evaluated in rats. After a 2-week pretreatment with BBR, the pharmacokinetic parameters of i.g. administered CBZ and ECBZ were not significantly altered. The pharmacokinetics of i.v. administered DIG was not modified by single and 2-week pretreatments with BBR, but a dose-dependent increase in AUC and C(max) was observed in the i.g. administered DIG parameters in rats. The AUCs of DIG with BBR (30 mg/kg, 100 mg/kg) were 133%, 170% (single) and 123%, 169% (2-week) of control, respectively. The AUC and C(max) of i.g. administered CsA with a 2-week pretreatment with BBR increased by 62% and 43% (BBR 30 mg/kg, p < 0.05), 96% and 60% (BBR 100 mg/kg, p < 0.01), compared with the control. In conclusion, berberine produced a dose-dependent increased bioavailability of digoxin and cyclosporine A by inhibition of intestinal P-gp. No significant changes in CYP3A activity by berberine were observed.

Retinoblastoma protein modulates gankyrin-MDM2 in regulation of p53 stability and chemosensitivity in cancer cells.

MDM2 is a key ubiquitin E3 ligase for p53 and its activity is critically regulated by a set of modulators, including ARF, p300, YY1 and recently by gankyrin, an oncoprotein frequently overexpressed in human heptocellular carcinomas. We have previously shown that MDM2 binds to and promotes retinoblastoma protein (Rb) degradation. Here we show that Rb inhibits MDM2 E3 ligase activity resulting in stabilization of p53. In addition, we demonstrated that Rb inhibits MDM2-mediated p53 ubiquitination in a gankyrin-dependent manner and the Rb-gankyrin interaction is critical for Rb-induced p53 stabilization. Furthermore, acute ablation of Rb facilitates gankyrin-mediated p53 destabilization, and desensitizes cancer cells for chemotherapy-induced apoptosis. These results indicate that Rb antagonizes gankyrin to inhibit MDM2-mediate p53 ubiquitination in cancer cells and suggest that the status of both p53 and Rb is important for efficacy of cancer chemotherapy.

Multivitamin use and B vitamin status in a homebound elderly population.

OBJECTIVE: Homebound elderly are at increased risk for micronutrient deficiencies and nutritional status in this population has not been adequately described. There is evidence for beneficial effects of multivitamin use and a greater understanding of their nutritional contribution could identify behaviors that may help alleviate excess chronic disease. The purpose of this analysis is to investigate, in a racially diverse group of homebound elders, the association of multivitamin use with measures of plasma B vitamin concentrations. DESIGN: We examined the cross-sectional association between multivitamin use and plasma concentrations of B vitamins and homocysteine in 236 white and 182 black homebound elders (65-99y). Dietary intake was assessed and demographic and health information was ascertained. RESULTS: White and black elders had a high prevalence of dietary intakes below the Estimated Average Requirement for folate (38.1 and 40.7%), vitamin B6 (16.9 and 19.2%.), and vitamin B12 (3 and 3.9%) respectively. Multivitamin use was associated with higher mean plasma B vitamin concentrations in each group. In whites, multivitamin users had higher concentrations of vitamin B6 (64.6 vs. 32.4 nmol/L; p < 0.001), vitamin B12 (398 vs. 324 pmol/L;p < 0.001) and folate (39.4 vs. 30.4 nmol/L;p < 0.001). Black multivitamin users had higher concentrations of vitamin B6 (53.7 vs. 29.5 nmol/L; p < 0.001), B12 (427 vs. 372 pmol/L; p < 0.05) and folate (35.7 vs. 25.4 nmol/L; < 0.001) than non-users. CONCLUSIONS: Multivitamin supplementation was associated with higher mean plasma concentrations of vitamins B6, B12, and folate and lower prevalence of low plasma B vitamin status in a biracial homebound elderly.

Structural basis of the multispecificity demonstrated by 17beta-hydroxysteroid dehydrogenase types 1 and 5.

17Beta-hydroxysteroid dehydrogenases/ketosteroid reductases (17beta-HSDs/KSRs) catalyze the last step of sex steroid synthesis or the first step of their degradation, and are thus critical for many physiological processes. The multispecificity demonstrated by 17beta-HSDs is important for steroid metabolism in gonadal and peripheral tissues, and is a consequence of the architecture of their binding and catalytic sites. Structurally, most of the family members are short chain dehydrogenase-reductases (SDRs) except the type 5 enzyme, which is an aldo-keto reductase (AKR). 17Beta-HSD type 1, a representative of the SDR family, has been studied extensively since the 1950s. However, its structure was not determined until the 1990s. It has always been considered as estrogen specific, in accord with the narrow binding tunnel that has been structurally determined and has been found to be complementary to estrogens. A recent study revealed that, in spite of the enzyme's narrow binding tunnel, the pseudo-symmetry of C19 steroids leads to its alternative binding, resulting in the multispecificity of the enzyme. Expressed in ovary, breast and placenta, the enzyme catalyzes the formation of another estrogen A-diol from DHEA in addition to the biosynthesis of estradiol; it also inactivates the most active androgen DHT by both 17beta-hydroxysteroid oxidation and 3-ketosteroid reduction. Type 5 17beta-HSD (AKR1C3) differs significantly from the type 1 enzyme by possessing a spacious and flexible steroid-binding site. This is estimated to be about 960 or 470 A3 in ternary complex with testosterone or 4-dione, respectively, whereas the binding site volume of 17beta-HSD1 is only about 340 A3. This characteristic of the 17beta-HSD5 binding site permits the docking of various steroids in different orientations, which encompasses a wider range of activities from 20alpha-, 17beta- and 3alpha-HSD/KSR to prostaglandin 11-ketoreductase. The in vitro activities of the enzyme are significantly lower than the type 1 enzyme. In the ternary complex with testosterone, the steroid C3-C17 position is quasi-reversed as compared to the complex with 4-dione. The multi-specificity contributes significantly to steroid metabolism in peripheral tissues, due to the high levels of 17beta-HSD5 mRNA in both breast and prostate tissues.

Characterization of binding of leukotriene C4 by human multidrug resistance protein 1: evidence of differential interactions with NH2- and COOH-proximal halves of the protein.

Multidrug resistance protein 1 (MRP1) is capable of actively transporting a wide range of conjugated and unconjugated organic anions. The protein can also transport additional conjugated and unconjugated compounds in a GSH- or S-methyl GSH-stimulated manner. How MRP1 binds and transports such structurally diverse substrates is not known. We have used [(3)H]leukotriene C(4) (LTC(4)), a high affinity glutathione-conjugated physiological substrate, to photolabel intact MRP1, as well as fragments of the protein expressed in insect cells. These studies revealed that: (i) LTC(4) labels sites in the NH(2)- and COOH-proximal halves of MRP1, (ii) labeling of the NH(2)-half of MRP1 is localized to a region encompassing membrane-spanning domain (MSD) 2 and nucleotide binding domain (NBD) 1, (iii) labeling of this region is dependent on the presence of all or part of the cytoplasmic loop (CL3) linking MSD1 and MSD2, but not on the presence of MSD1, (iv) labeling of the NH(2)-proximal site is preferentially inhibited by S-methyl GSH, (v) labeling of the COOH-proximal half of the protein occurs in a region encompassing transmembrane helices 14-17 and appears not to require NBD2 or the cytoplasmic COOH-terminal region of the protein, (vi) labeling of intact MRP1 by LTC(4) is strongly attenuated in the presence of ATP and vanadate, and this decrease in labeling is attributable to a marked reduction in LTC(4) binding to the NH(2)-proximal site, and (vii) the attenuation of LTC(4) binding to the NH(2)-proximal site is a consequence of ATP hydrolysis and trapping of Vi-ADP exclusively at NBD2. These data suggest that MRP1-mediated transport involves a conformational change, driven by ATP hydrolysis at NBD2, that alters the affinity with which LTC(4) binds to one of two sites composed, at least in part, of elements in the NH(2)-proximal half of the protein.

Growth and development in term infants fed long-chain polyunsaturated fatty acids: a double-masked, randomized, parallel, prospective, multivariate study.

OBJECTIVE: To evaluate the effects of dietary intake of the long-chain polyunsaturated fatty acids, arachidonic acid (AA), and docosahexaenoic acid (DHA) on multiple indices of infant growth and development. DESIGN: A double-masked, randomized, parallel trial was conducted with term infants fed formulas with or without AA+DHA for 1 year (N = 239). Reference groups of breastfed infants (N = 165) weaned to formulas with and without AA+DHA were also studied. Infants in the formula groups were randomized at

Isolation, characterization, and mapping of a novel human KRAB zinc finger protein encoding gene ZNF463.

A novel human KRAB (Krüppel associated box) type zinc finger protein encoding gene, ZNF463, was obtained by mRNA differential display and RACE. It consists of 1904 nucleotides and encodes a protein of 463 amino acids with an amino-terminal KRAB domain and 12 carboxy-terminal C2H2 zinc finger units. The gene is mapped to chromosome 19q13.3 approximately 4 by FISH. As from Northern blot analysis ZNF463 is only expressed in testis, RT-PCR indicates that ZNF463 is expressed more highly in normal fertile adults than in fetus and azoospermic patients suggesting that it may play a role in human spermatogenesis.

Outgrowth of chondrocytes from human articular cartilage explants and expression of alpha-smooth muscle actin.

The objectives of this study were to investigate the effect of various enzymatic treatments on the outgrowth of chondrocytes from explants of adult human articular cartilage and the expression of a specific contractile protein isoform, alpha-smooth muscle actin, known to facilitate wound closure in other connective tissues. Explants of articular cartilage were prepared from specimens obtained from patients undergoing total joint arthroplasty. The time to cell outgrowth in vitro was determined and the expression of alpha-smooth muscle actin shown by immunohistochemistry. Treatment of the explants with collagenase for 15 minutes reduced the time to outgrowth from more than 30 days to 3 days. Hyaluronidase, chondroitinase ABC, and trypsin applied for the 15-minute period had no effect on the time to cell outgrowth when compared with untreated controls. Pretreatment with hyaluronidase prior to collagenase reduced the time to outgrowth. A notable finding of this study was that the majority of chondrocytes in the adult human articular cartilage specimens and virtually all of the outgrowing cells contained alpha-smooth muscle actin. We conclude that human articular chondrocytes have the capability to migrate through enzymatically degraded matrix and express a contractile actin isoform. Collagenase treatment reduces the time required for cell outgrowth.

[Effects of maxillary growth of Wistar rats with bilateral artificial cleft palate after premaxillary orthopedic treatment on expression of proliferating cell nuclear antigen(PCNA)].

OBJECTIVE: Being based on the principle of Latham's appliance, this experiment is designed to detect activity of osteoblasts in the maxillary sutures of Wistar rats with bilateral artificial cleft palate by immunohistochemistry technique, and evaluate the effects of this appliance on the growth and development of the maxilla. METHODS: The animal models of male infant Wistar rats with bilateral cleft palate were made by removing some palatal bones, splitting a "V" gap of 1.5 cm wide at the line between premaxilla and segments of maxilla. A sort of appliances, which could be fixed in the mouth of Wistar rats with bilateral cleft palate for correcting protrusion premaxilla was constructed basing on the principle of Latham's appliance. Then the diferent pathological changes of osteoblast proliferation between the experimental group and the two controlled groups were examined. The jugomaxillary sutrues, temporomalar sutures and sphenoipalatine suture were harvested 7, 14 days after premaxillar orthopedic treatment, followed by 4% paraformaldehyde fixing about 1 hour, demineralization with 15% EDTA and 0.5% paraformaldehyde for 48 hours, distilled water washing for a night, dimethylbenzene transparant dealing, and paraffin wax embeding. Proliferating Osteoblasts in all these sutures were investigated using immunohiostochemical technique with monoclonal antibodies of proliferating cell nuclear antigen (PCNA). RESULTS: Seven days after orthopedic treatment, no significant difference was observed between the experimental group and the two controlled groups. While after fourteen days, obvious PCNA-positive expression were observed in cells of all these sutures of the experimental group. CONCLUSION: The distribution of proliferating cells and the degree of cell proliferation change after premaxillary orthopedic treatment. And significant cell proliferation is observed in the experimental group, but there are no significant differences between the two controlled groups.

Murine p38-delta mitogen-activated protein kinase, a developmentally regulated protein kinase that is activated by stress and proinflammatory cytokines.

The p38 mitogen-activated protein kinases (MAPK) play a crucial role in stress and inflammatory responses and are also involved in activation of the human immunodeficiency virus gene expression. We have isolated the murine cDNA clones encoding p38-delta MAPK, and we have localized the p38-delta gene to mouse chromosome 17A3-B and human chromosome 6p21.3. By using Northern and in situ hybridization, we have examined the expression of p38-delta in the mouse adult tissues and embryos. p38-delta was expressed primarily in the lung, testis, kidney, and gut epithelium in the adult tissues. Although p38-delta was expressed predominantly in the developing gut and the septum transversum in the mouse embryo at 9.5 days, its expression began to be expanded to many specific tissues in the 12.5-day embryo. At 15.5 days, p38-delta was expressed virtually in most developing epithelia in embryos, suggesting that p38-delta is a developmentally regulated MAPK. Interestingly, p38-delta and p38-alpha were similar serine/threonine kinases but differed in substrate specificity. Overall, p38-delta resembles p38-gamma, whereas p38-beta resembles p38-alpha. Moreover, p38-delta is activated by environmental stress, extracellular stimulants, and MAPK kinase-3, -4, -6, and -7, suggesting that p38-delta is a unique stress-responsive protein kinase.

[Detection of clinical diagnosis and different symptoms of Sjogren's syndrome by traditional chinese medicine (TCM)]

OBJECTIVE:To search for the appropriate diagnostic criteria of SS and confirm the differentiation of SS.METHODS:The European di agnostic criteria has been used in clinic work for about two years,and differentiated the symptoms of SS by TCM. The specialty and relationship between them was made an inquiry.RESULTS:The European criteria set for SS offers the clinic work a practical method to correctly classify the patients suspected of having this disorder. It refers to the signs and symptoms caused by the insufficiency of spleen-qi with deficiency of liver-yin and kidney-yin.CONCLUSION:It has been further proved for easy to make up the diagnostic of SS and its treatment using traditional chinese medicine.

[Experimental research on animal model and treatment of Sjogren's syndrome by traditional chinese medicine (TCM)]

OBJECTIVE:To confirm the function of SS Syrup used in our department. METHODS: Under the basis of TCM and differentiation symptoms of SS,the animal model was established and applied to test the function of SS syrup.There are three indices to be observed,weight change,volume of drinking water daily,cAMP change. RESULTS: The three observed indices of dificiency symptom model had been rectified by SS syrup. CONCLUSION: SS syrup has two main functions:(1)reinforcing of the spleen and tonifying qi;(2)nourishing liver-yin and kidney-yin as well as removing pathogentic heat and fire.

Clinical interpretations of transient otoacoustic emissions.

PURPOSE: The purpose of this study was to characterize the relation of different ordinal patterns of transient otoacoustic emissions (TEOAES) with respect to underlying otologic disorders and auditory status. PATIENTS AND METHODS: The results of TEOAEs in 225 patients with various auditory disorders were investigated and compared with normative data established from 90 subjects of various ages. TEOAEs were categorized according to four patterns: (1) normal (general response level within 90% of normative data, (2) reduced amplitude (general response level was > or =2 dB peak sound pressure level (pSPL), but less than the mean -1.64 SD of the normative data), (3) abnormal morphology of frequency spectrum (general response level was within normal limits, but reduced at > or =2 individual octave frequencies between 1,000 and 5,000 Hz), and (4) total absence (response level <2 dB pSPL). RESULTS: This study showed that the normal pattern of TEOAEs, in terms of response amplitude, varied with age. Our results further indicated that a reduced amplitude pattern of TEOAEs was noted in patients with a mild sensorineural hearing loss (SNHL), negative tympanometric pressure, a pressure-equalization tube, and Meniere's disease. TEOAEs provided good frequency-specific information for patients with a noise-induced hearing loss. All patients with ossicular chain abnormalities, more than moderate SNHL, and a middle ear mass or effusion had total absence of TEOAEs. Patients with acoustic neuroma and brainstem lesions presented a complex profile of TEOAEs. In the follow-up of auditory function in patients undergoing otologic surgery, different patterns of TEOAEs between the preoperative and postoperative recordings were evident, which correlated with the hearing thresholds and middle ear status. The abnormal findings of TEOAEs due to specific auditory diseases were discussed. CONCLUSION: The interpretation of TEOAEs can be facilitated through an analysis of specific patterns and in combination with other audiologic tests.

Current evaluation of pseudohypacusis: strategies and classification.

Some cases of pseudohypacusis may involve medicolegal aspects and require a confirmed and quantitative diagnosis. These challenging cases must be identified, and then evaluated with basic audiologic and sophisticated electrophysiologic tests. Data on 64 patients with pseudohypacusis collected over a 4-year period are reported. A classification system was developed from an analysis of these cases and is presented for clinical evaluation and diagnosis. In many cases, conventional audiologic evaluation involving pure tone and speech audiometry may be adequate and sufficient for diagnosis. In more complex cases, evoked otoacoustic emissions (EOAEs) and auditory brain stem responses (ABRs) are needed for confirmation of peripheral auditory sensitivity. We found that EOAEs were the most rapid economical, and objective method, and confirmed the diagnosis of hearing loss in 78.1% of cases. Fifteen percent of subjects required ABRs to substantiate the diagnosis. The reliability of basic audiologic tests based on previous clinical investigations and data from the literature are discussed. We conclude that a thorough knowledge and understanding of pseudohypacusis is essential to verify the existence of pseudohypacusis, to determine its type, and to quantify the auditory thresholds.

[A randomized clinical trial for the evaluation of head and neck squamous cell carcinoma treating with traditional Chinese medicine]

This paper described the research design,research process and statistical analysis of randomized clinical trial by treatment with traditional Chinese medicine in patients with squamous cell carcinoma of the head and neck.At first we found there was no significant difference between treating group and contral group(P>0.05) using log-rank test on Kaplan-Meier survival curves. With further multivariate analysis Cox model and after adjusting for sex,age,legth of taking the assigned medication,grade of tumor differentiation and using of chemotherapy,the rate of tumor-related death for control group was significantly higher than that for treating group :hazards ratio estimate was 1.939(95% confidence interval 1.008-3.729,(P<0.05).The length of taking Chinese medicine was one of the most important prognostic factors(P<0.001).The results suggest that the prescribed medication contributed to the improved survival,and it was an independent prognostic factor.

[Hyperlipidemia treated with xiaobu jianfei pian].

A total of 51 cases with hyperlipidemia, who were defined deficiency symptom-complex complicated by symptoms of excessiveness in TCM were studied clinically. The patients were divided into two groups at random. One group was treated with Xiaobu Jianfei Pian (XJP) as treated group, another with Fangfeng Tongsheng San as a control. It was found that XJP was able to lower total serum cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and apolipoprotein (apo) B significantly (P less than 0.001, 0.001, 0.001) while it had markedly improved clinical symptoms. It was also observed that XJP had good effects on the ratios of apoA1/B and TC/HDL-C, and was able to reduce body weight index. All of these were better than those of the control group statistically. These evidences indicate that XJP possesses clinical therapeutic effects on both lipid-lowering and lipid-adjusting, which suggest that XJP may be an effective anti-hyperlipidemia medicine.