Measurement of α-dicarbonyl compounds in human saliva by pre-column derivatization HPLC.

Background α-Dicarbonyl compounds (α-DCs) have been detected in body fluids including plasma and urine and elevation of this sort of compounds in vivo has been associated with the development of many kinds of chronic diseases. However whether α-DCs are present in human saliva, and if their presence/absence can be related with various chronic diseases is yet to be determined. Methods In this study, a pre-column derivatization HPLC-UV method was developed to measure 3-deoxyglucosone (3-DG), glyoxal (GO), methylglyoxal (MGO), diacetyl (DA), and pentane-2,3-dione (PD) in human saliva employing 4-(2,3-dimethyl-6-quinoxalinyl)-1,2-benzenediamine (DQB) as a derivatizing reagent. The derivatization of the α-DCs is fast and the conditions are facile. The method was evaluated and the results show that it is suitable for the quantification of α-DCs in human saliva. Results In the measurements of these α-DCs in the saliva of 15 healthy subjects and 23 type 2 diabetes mellitus (T2DM) patients, we found that the concentrations of GO and MGO in the saliva of the diabetic patients were significantly higher than those in healthy subjects. As far as we know, this is the first time that salivary α-DC concentrations have been determined and associated with T2DM. Conclusions The developed method would be useful for the measurement of the salivary α-DC levels and the data acquired could be informative in the early screening for diabetes.

Polymorphisms of 23 Y-chromosome STR Loci in A Han Population in Jiangsu Province.

Objective To investigate the polymorphisms of 23 Y-STR loci in a Han population in Jiangsu province. Methods Blood samples were collected from 4821 unrelated healthy Han males in Jiangsu province. DNA templates were amplified by PowerPlex Y23 kit,and the amplification products were detected by 3500xL genetic analyzer. Then,we calculated the allele frequencies and gene diversities respectively,as well as the haplotype frequencies and haplotype diversities. Results The gene diversity of these 23 Y-STR loci ranged 0.4099-0.9696. A total of 4781 haplotypes were detected,of which 4743 were found once. The haplotype diversity was 0.99999812. Conclusion The 23 Y-STR loci used in this study are highly polymorphic in Han individuals in Jiangsu province and therefore suitable for population genetic study and forensic individual identification.

Human amniotic epithelial cells combined with silk fibroin scaffold in the repair of spinal cord injury.

Treatment and functional reconstruction after central nervous system injury is a major medical and social challenge. An increasing number of researchers are attempting to use neural stem cells combined with artificial scaffold materials, such as fibroin, for nerve repair. However, such approaches are challenged by ethical and practical issues. Amniotic tissue, a clinical waste product, is abundant, and amniotic epithelial cells are pluripotent, have low immunogenicity, and are not the subject of ethical debate. We hypothesized that amniotic epithelial cells combined with silk fibroin scaffolds would be conducive to the repair of spinal cord injury. To test this, we isolated and cultured amniotic epithelial cells, and constructed complexes of these cells and silk fibroin scaffolds. Implantation of the cell-scaffold complex into a rat model of spinal cord injury resulted in a smaller glial scar in the damaged cord tissue than in model rats that received a blank scaffold, or amniotic epithelial cells alone. In addition to a milder local immunological reaction, the rats showed less inflammatory cell infiltration at the transplant site, milder host-versus-graft reaction, and a marked improvement in motor function. These findings confirm that the transplantation of amniotic epithelial cells combined with silk fibroin scaffold can promote the repair of spinal cord injury. Silk fibroin scaffold can provide a good nerve regeneration microenvironment for amniotic epithelial cells.

Systemic evaluation of hypoxic-ischemic brain injury in neonatal rats.

The objective of the study was to evaluate the systematically rat model of neonatal hypoxic-ischemic brain damage. The right carotid arteries of 7-day-old healthy Wistar rats were ligated, and then, the rats were subjected to an environment with 8 % of oxygen. Four weeks after the birth, neurobehavioral test, water maze test, and motor-evoked potential and neuropathologic examinations were performed. The footprint analysis showed significantly larger and instable paces in the hypoxic-ischemic group (P < 0.05); the time that rats crossed the balance beam in the hypoxic-ischemic group was longer than the control group (P < 0.05). The water maze test showed that the escape latency of hypoxic-ischemic group was significantly longer than that of control group (P < 0.05). The hindlimb quadriceps compound muscle-evoked potential CMEP of rats in hypoxic-ischemic group showed that the wave amplitude was lower than that of control group (P < 0.05). HE staining showed visible periventricular leukomalacia in hypoxic-ischemic groups; disrupted nuclear membrane was detected in the IH group with transelectronmicroscopy; Immunohistochemistry: compared with control group, MBP-positive neurocytes decreased, glial fibrillary acidic protein positive neurocytes increased in the periventricular zone (P < 0.05). Carotid artery ligation combining the hypoxic chamber created a reliable and stable rat model of neonatal hypoxic-ischemic brain damage and can be used for experimental research related to management of cerebral palsy.

[Intranasal vaccination with mycobacterial 65-kD heat-shock protein can prevent insulitis and diabetes in non-obese diabetic mice].

AIM: To study the efficacy of heat shock protein 65 kDa (HSP65) of Mybobacterium tuberculosis var. bovis in prevention of autoimmune diabetes by intranasal. METHODS: The HSP65 gene was derived from Mybobacterium tuberculosis var. bovis genome by PCR and successfully expressed as soluble protein in Escherichia coli. The recombinant protein HSP65 was purified by anion exchange column chromatography, then used to immunize prediabetic NOD (non-obese diabetic) mice via three intranasal (i.n.) delivery in absence of adjuvants. Serum samples from the immunized mice were collected at monthly intervals. The anti-HSP65 antibody was detected by enzyme-linked immunosorbent assay (ELISA) and verified by Western blot analysis. The concentration of blood glucose was measured by automatic analyzer. RESULTS: Specific anti-HSP65 antibodies were successfully induced in mice immunized via intranasal routes. Histochemical analysis of mice pancreas tissue showed that HSP65 intranasal vaccination could decrease pathological changes in NOD mice. CONCLUSION: Intranasal vaccination with HSP65 in NOD mice could prevent the development of diabetes. Our results demonstrate that intranasal vaccination with HSP65 reduces significantly the inflammatory process associated with auto-immune diabetes. This approach may offer novel therapeutic avenues for the treatment for of type 1 diabetes mellitus.

[Clinical study on a randomized, double-blind control of Shenwu gelatin capsule in treatment of mild cognitive impairment].

OBJECTIVE: To evaluate the effectiveness of Shenwu gelatin capsule (compound Chinese ginseng and fleeceflower root etc) in the treatment on senile mild cognitive impairment. METHOD: 166 patients meeting criteria for mild cognitive impairment were selected from patients in Dongzhimen Hospital and were randomly assigned into a treatment group (n = 83 cases), given 5 capsules of Shenwu gelatin capsule, and a positive control group (n = 83 cases) , given 2 capsules of aniracetam with 3 placebo capsules. All subjects took this medication 3 times a day for 3 months, double-blind and double-moulding control were used in this study. At a baseline and end (three months later), all subjects were assessed using a battery consisting of MMSE and clinical memory scale (CMS). RESULT: Both Shenwu gelatin capsule and aniracetam can remarkably increase the score of memory quotient (MQ), P < 0. 01. They have the similarly effectiveness, there are no statistical difference between the two groups in effectiveness of increasing memory scores. CONCLUSION: Shenwu gelatin capsule has certain effects on mild cognitive impairment.

[Comparative research of the influence factors of DNA extraction of bloodstain on the filter paper with Chelex-100 method].

OBJECTIVE: To improve DNA extraction from bloodstain on the filter paper and to establish a rapid, simple, and cost-effective method for DNA extraction suitable for database construction. METHODS: Seven hundred and fifty two aged bloodstains on filter paper were randomly divided into four groups. The four different DNA extraction methods were compared with each other, and two DNA extraction methods used for 63 fresh bloodstains on filter paper were also compared with each other. RESULTS: There were no statistically significant differences observed among the four DNA extraction methods (P > 0.05) for aged bloodstains on filter paper; But the difference between the two DNA extraction methods for fresh bloodstains on filter paper was obviously (P < 0.05). CONCLUSION: Extraction of DNA samples from aged bloodstains on filter paper can be accomplished by using Chlex-100 methodology directly with no need to wash the bloodstains.

[A clinical study on a randomized, double-blind control of Chinese medicine granules in treatment of vascular dementia].

OBJECTIVE: To evaluate the effectiveness of Chinese medicine granules (Compound Chinese extract from herbs) in the treatment on senile vascular dementia. METHOD: One hundred and twenty patients meeting criteria for vascular dementia were selected from patients in Dongzhimen Hospital and were randomly assigned into a treatment group (n = 70 cases), given 1 package of Chinese medicine granules with 1 placebo tablets, and a positive control group (n = 50 cases), given 1 tablets of Duxil with 1 package of placebo. All subjects took this medication 3 times a day for 2 months. Double-blind and double-moulding control were used in this study. At a baseline and end (two months later), all subjects were assessed using a battery consisting of MMSE and Blessed behavior measuring scale. RESULT: Both Chinese medicine granules and Duxil could remarkably increase the score of cognition and activity (P < 0.01). They had Similarly effectiveness and there was no statistical difference between the two groups in effectiveness of increasing memory scores. Chinese medicine granules was better than Duxil in increasing the scores of behavior (P < 0.05). CONCLUSION: Chinese medicine granules has certain effects on vascular dementia. And it has remarkable effectiveness in ameliorating the status of total body. And it can relieve the symptoms in vascular dementia.

[The influence of CGE on expression of IL-1beta and c-fos protein in the hippocampus region in rats following cerebral ischemia-reperfusion].

OBJECTIVE: To observe inhibiting effect of CGE (compound ginseng extract) on increased expression of IL-1beta and c-fos protein following cerebral ischemia-reperfusion. METHOD: The vascular dementia model was made by middle cerebral artery occlusion for 2 hours. Expression of IL-1beta and c-fos were determined by immunohistochemistry in the hippocampus regions in brain tissue at the 0.5 h-7 d after reperfusion. CGE was diluted by CMC and poured into the stomach by 0.7 mL x (100 g)(-1) with a high dosage (19.34 x 10(3) g x L(-1) row herbs), a middle dosage (9.67 x 10(3) g x L(-1)), a low dosage (4.83 x 10(3) g x L(-1)). There were an IL-1ra (rhIL-1ra 20 microg injected into the left cerebral ventricle), a sham operation (NaCl 20 microL injected into the left cerebral ventricle) and a model as control. RESULT: Compared with control group, three dose groups (low, middle and high) in CGE showed significant inhibiting effects on the expression of c-fos protein at 2, 3, 4, 12 hours and 3 day following cerebral ischaemic-reperfusion. The level of the inhibiting effects in small and middle groups were lower at all time points than that in IL-1ra group (P < 0.05 to P < 0.01). CGE inhibited the expression of hippocampus IL-1beta protein, taking effect from the 2 h after reperfusion. Both HD group (531 +/- 151.1) and MD group (589.3 +/- 78.6) showed more obvious effect which lasted until the 72 h compared with the model group (687.6 +/- 116.7) (P < 0.01 and 0.05). Large dose group (81.3 +/- 16.1) showed the same level of the inhibiting effect on expression of c-fos protein as IL-1ra group (67.2 +/- 25.7) from 4 hour on following cerebral ischaemic reperfusion (P > 0.05). CONCLUSION: CGE with function of Yiqi Bushen, Huoxue Huatan has effect of inhibiting up-regulated expression of IL-1beta and c-fos protein following cerebral ischemia-reperfusion. However, this effect of CGE starts relatively later than that of IL-1ra. The effect of CGE is associated with its dosage, i.e. a larger dosage has a better effect on expression of c-fos protein in post-stroke dementia.

[A follow-up study on a randomized, single-blind control of King's Brain pills in treatment of memory disorder in elderly people with MCI in a Beijing community].

OBJECTIVE: To evaluate the effect of King's Brain pills (Compound Chinese ginseng extract from herbs) on the treatment and the delaying of memory decline in the elderly with mild cognitive impairment (MCI) in a community by a year follow-up of neuropsychology. METHOD: 75 patients with MCI were selected from Beixinqiao community of Beijing by a cutoff score of 0.0/0.5 on CDR and were randomly assigned into a treatment group (n = 30 cases), given 4 pills of a compound Chinese ginseng extract (King's Brain) with 2 placebo tablets, and a positive control group (n = 30 cases), given 2 tablets of Piracetam with 4 placebo tablets, as well as a placebo group (n = 15 cases), given a placebo of 4 tablets and 2 pills. All subjects took this medication 3 times a day for 3 months. Single-blind and double-moulding control were used in this study. At a baseline and a middlepoint (after 3 months), and a follow-up end (one year later) following a three months of medication therapeutics, all subjects were assessed using a battery consisting of MMSE and 5 memory items on BNPT battery. RESULT: In the treatment group, MMSE score 27.50 +/- 1.68 was increased to 28.27 +/- 1.70 after 3 months but decreased to 26.90 +/- 1.90 after one year of the treatment. However, the latter score was higher than that in a placebo group 26.33 +/- 1.03 (P < 0.05). Verbal Learning Test score was significantly increased from 68.73 +/- 28.74 at baseline to 87.33 +/- 29.78 at follow-up point in the treatment group, which was significantly higher than that in the placebo group (P < 0.01). The total score of memory items on BNPT battery was significantly increased from 78.23 +/- 28.98 at baseline to 93.53 +/- 35.56 at follow-up point in the treatment group, higher than that in Piracetam group (P < 0.05) and the placebo group (P < 0.01). CONCLUSION: Both King's Brain spills and piracetam tablets have protective effect on cognitive and memory decline in elderly with MCI.