[Linkage analysis of susceptibility loci in 2 target chromosomes in pedigrees with paranoid schizophrenia and undifferentiated schizophrenia].

OBJECTIVE: To investigate the relationship of susceptibility loci in chromosomes 1q21-25 and 6p21-25 and schizophrenia subtypes in Chinese population. METHODS: A genomic scan and parametric and non-parametric analyses were performed on 242 individuals from 36 schizophrenia pedigrees, including 19 paranoid schizophrenia and 17 undifferentiated schizophrenia pedigrees, from Henan province of China using 5 microsatellite markers in the chromosome region 1q21-25 and 8 microsatellite markers in the chromosome region 6p21-25, which were the candidates of previous studies. All affected subjects were diagnosed and typed according to the criteria of the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revised (DSM-IV-TR; American Psychiatric Association, 2000). All subjects signed informed consent. RESULTS: In chromosome 1, parametric analysis under the dominant inheritance mode of all 36 pedigrees showed that the maximum multi-point heterogeneity Log of odds score method (HLOD) score was 1.33 (α = 0.38). The non-parametric analysis and the single point and multi-point nonparametric linkage (NPL) scores suggested linkage at D1S484, D1S2878, and D1S196. In the 19 paranoid schizophrenias pedigrees, linkage was not observed for any of the 5 markers. In the 17 undifferentiated schizophrenia pedigrees, the multi-point NPL score was 1.60 (P= 0.0367) at D1S484. The single point NPL score was 1.95(P= 0.0145) and the multi-point NPL score was 2.39 (P= 0.0041) at D1S2878. Additionally, the multi-point NPL score was 1.74 (P= 0.0255) at D1S196. These same three loci showed suggestive linkage during the integrative analysis of all 36 pedigrees. In chromosome 6, parametric linkage analysis under the dominant and recessive inheritance and the non-parametric linkage analysis of all 36 pedigrees and the 17 undifferentiated schizophrenia pedigrees, linkage was not observed for any of the 8 markers. In the 19 paranoid schizophrenias pedigrees, parametric analysis showed that under recessive inheritance mode the maximum single-point HLOD score was 1.26 (α = 0.40) and the multi-point HLOD was 1.12 (α = 0.38) at D6S289 in the chromosome 6p23. In nonparametric analysis, the single-point NPL score was 1.52 (P= 0.0402) and the multi-point NPL score was 1.92 (P= 0.0206) at D6S289. CONCLUSION: Susceptibility genes correlated with undifferentiated schizophrenia pedigrees from D1S484, D1S2878, D1S196 loci, and those correlated with paranoid schizophrenia pedigrees from D6S289 locus are likely present in chromosome regions 1q23.3 and 1q24.2, and chromosome region 6p23, respectively.

[Systemic oxidative stress induced by intratracheal instilling with PM10 in rats].

OBJECTIVE: To observe the oxidative stress of serum, lung and heart tissues in rats caused by PM10. METHODS: In the study,24 male Wistar rats weighed 320-360 g were randomly divided into four groups, including one saline control group and three PM10 exposed groups. PM10 was administered to the exposed groups by intratracheal instillation at the doses of 3.75, 7.5, 15 mg/kg, respectively. The rats were sacrificed 24 hours after exposure. The levels of MDA and activities of SOD in the serum, lung and heart tissues were measured. In addition, the contents of protein carbonyl in lung and heart tissues were determined. RESULTS: The activities of SOD in the serum, lung and heart tissues decreased markedly in the 7.5 and 15 mg/kg groups. The levels of MDA in the lung tissues significantly increased in the 7.5 and 15 mg/kg groups, while those in the serum and heart tissues significantly increased only in the 15 mg/kg group. The contents of protein carbonyl in the lung tissues markedly increased in the 15 mg/kg group, but no significant change was found in the heart tissues. CONCLUSION: The study showed that PM10 can cause oxidative stress effects in rats. The lipids of cell membrane are more susceptible to oxidative stress than the proteins.

Nanoemulsion-based delivery system for enhanced oral bioavailability and caco-2 cell monolayers permeability of berberine hydrochloride.

Berberine hydrochloride (BBH) has a variety of pharmacological activities such as antitumor, antimicrobial, anti-inflammation, and reduce irritable bowel syndrome. However, poor stability and low oral bioavailability limited its usage. Herein, an oil-in-water nanoemulsion system of BBH was developed to improve its stability and oral bioavailability. The pseudoternary phase diagrams were constructed for the determination of composition of various nanoemulsions. The nanoemulsions of BBH composed of Labrafil M 1944 CS (oil phase), RH-40 (surfactant), glycerin (co-surfactant), and water (aqueous phase). The O/W nanoemulsion of BBH showed a relative bioavailability of 440.40% compared with unencapsulated BBH and was stable in our 6-month stability study. Further, there was a significant increase in intestinal permeability of BBH as assessed by Caco-2 cell monolayers and a significant reduction in efflux of BBH by the multidrug efflux pump P-glycoprotein. This study confirmed that the nanoemulsion formulation could be used as an alternative oral formulation of BBH to improve its stability, oral bioavailability and permeability.

[Development and challenge of modified hemoglobins as red blood cell substitutes].

The problems of blood shortage and the virus infection risk of blood transfusion have promoted the study of blood substitutes. Modified hemoglobin has become the focus of the challenges research because of its excellent oxygen carrying ability. To overcome the toxicity effect on direct use of purified native hemoglobin, various modification technologies have been developed, including diaspirin cross-linking, glutaraldehyde polymerization, O-raffinose polymerization, polyethylene glycol conjugation, liposome encapsulation and biodegradable polymer encapsulation. Some of the products have been in clinical trials, and one of the products has been approved in a country for clinical use. Research on red blood cell substitutes in China has also developed fast. This paper provides an overview of the history and current status in development of different hemoglobin-based red blood cell substitutes, especially the problems encountered, the challenges faced, and the prospects in future.

[Simultaneous purification of superoxide dismutase, catalase and hemoglobin from bovine erythrocyte lysate with PEG 600 as chaperon].

Superoxide dismutase, catalase and hemoglobin were purified simultaneously from the same batch of bovine erythrocyte lysate. The process involves an initial anion exchange chromatography, followed by a hydrophobic interaction chromatography and gel filtration chromatography. 0.75% polyethylene glycol 600 was added as a purification chaperon before the anion exchange chromatography. The hemoglobin fraction passed through the ion exchange column without being retained. The superoxide dismutase and catalase were adsorbed by the column and were eluted separately during elution. The two eluted fractions containing crude superboxide dismutase and catalase were further purified with hydrophobic interaction chromatography and gel filtration chromatography in sequence. The specific activities of superoxide dismutase and catalase were 15932u/mg and 65918u/mg, respectively. SDS-polyacrylamide gel electrophoresis and gel filtration chromatography were used to analyze the purity of the proteins. The purity of superoxide dismutase, catalase and hemoglobin were 77.6%, 81.9% and 99.9%, respectively. The total recoveries for superoxide dismutase, catalase and hemoglobin were 47.4%, 29.6% and 88.7%, respectively.

Conjugate of bovine hemoglobin and human serum albumin as a candidate for blood substitute: characteristics and effects on rats.

Conjugate of bovine hemoglobin (bHb) and human serum albumin (HSA) was prepared. The product was simply composed of 89.7% one-to-one Hb-HSA conjugate, 6.0% oligomer of Hb and HSA, 3.5% unconjugated HSA and 0.8% unconjugated Hb, with an average molecular weight of 157 kD. The physicochemical characteristics were determined. Effects of single replacement on blood pressure and long-term survival of rats with 30% and 60% acute blood loss were studied, in comparison with Ringer-lactate solution, stroma-free hemoglobin (SFHb), 5% HSA in Ringer-lactate, whole blood and no resuscitation fluid. Results showed that Hb-HSA conjugate maintained the mean arterial pressure of rats to initial level with no pressor effect. Long-term effects of the replacement fluids on 30% bleeding rats showed that, for the group infused with Hb-HSA conjugate, histology of five major organs, heart, kidney, liver, spleen and lung, were essentially normal, similar to that of whole blood, while obviously renal side-effects appeared in other groups. The efficacy of the conjugate was further demonstrated by the resuscitation of lethal hemorrhagic shock rats (60% acute blood loss) with 100% survival rate (followed for 14 days), the same result as whole blood. The Hb-HSA conjugate can thus be another candidate for blood substitute in emergency.

[Progress in purification of human serum albumin].

Human serum albumin(HSA) has been used clinically to treat a number of diseases with high dosage. Extremely pure puoduct is required in large-scale production. Plasma-derived HSA(pHSA) has long been produced by precipitation methods. Among them cold ethanol precipitation is dominant. However, chromatographic purification of HSA has been increasingly studied in the last few years. Application of chromatography, especially ionexchange, affinity, and size-exclusion, has opened a new area in the production of pHSA. A new challenge is the purification of recombinant HSA(rHSA). A successful approach involves STREAMLINE expanded bed adsorption to direct capture the target product from the fermentation broth. This novel process eliminates the need to separate the cells by centrifugation or membrane filtration. Ion exchange chromatography and hydrophobic chromatography play a central role in the purification scheme. Integration with other chromatographic techniques such as size-exclusion, metal chelate, and affinity gives improved purification results. Though innovative, the purification of rHSA still needs further improvement and optimization to increase product purity and process recovery.