Objective To examine the effect of Caffeine on the cultured cortical neuron apoptosis in neonatal rats.Methods The primary cerebral cortex neurons for cultures were obtained from neonatal mice 2-3 days after birth,Caffeine reconstituted at final concentrations 300μmol/L and 1 000μmol/L was added to the cell cultures and continuously co-incubated for 6-36 h,respectively after the cortical neurons were continuously cultivated 7 days after incubation under temperature of 37℃ incubator with 5% CO_2 and 100% relative humidity,the intracellular calcium concentration,mitochondrial membrane potential and apoptosis rate were determined by the flow cytometry.The activity of Caspase-9 was assayed by enzyme-labeled instrument,and Caspase-9 activity by the enzyme-1inked analyzer.Cell morphological changes were observed under electron microscope and fluorescent microscope after being stained with Hoechst 33258 fluorescent dye.Results Compared with the control group,the average increase in intracellular calcium fluorescence intensity was most significant(P<0.05),which elevated from the normal value 43.13±2.02 to 45.28±1.16 and 46.92±1.99,respectively at 6 h;mitochondrial membrane potentials were reduced most significandy(P<0.05).from the base value 443.58 ±11.77 down to 289.53±16.47 and 165.14±14.72,respectively at 8h.Caspase-9 activity was peaked(P<0.05),from the normal value 1.00±0.000 to 5.33±1.02 and 8.33±0.92,respectively at 10 h.The neuronal apoptosis ratio was increased significantly (P<0.05),from the normal value 4.94±1.74 to 15.98±2.03 and 18.70±2.09,at 36h.The apoptotic bodies were observed at 24 h after administration of 300 μmol/L and 1000 μmol/L Caffeine.Conclusion Caffeine may promote neuronal apoptosis in neonate mice.

Microimmunoperoxidase (Micro-IP) test was established for detection of IgM and IgG antibodies of typhus fever. 71 cases with clinically diagnosed typhus fever were examined and the positive rate was 86.3% for paired sera and 60.0% for single sera. The results coincided with those of Micro-IF in all cases, IgM can be detected as early as 4th day after onset and persisted as long as 9 months. 76.2% sera showed positive reaction in the first week post onset. Micro-IP showed group specificity to sera from immunized rabbits and the patients with typhus fever. The advantages of Micro-IP were simple, rapid, sensitive, specific and economic for reagents and time.

Objective To determine the influence of intense pulsed light (IPL) on the secretion of TGF-β1 in cultured human fibroblasts and the intervention of JNK inhibitor.Methods The callan foreskin fibroblasts were cultured and divided into 2 groups. In the IPL treatment group, cells were irradiated with IPL with fluences of0 (negative control), 10, 18, 27, 36, and 36 J/cm2 × 2 (irradiated with IPL with fluences of 36 J/cm2 twice). In the IPL + inhibitor group, cells were irradiated with IPL with fluences of 36 J/cm2 after incubation with the inhibitor SP600125 for 2 h. TGF-β1 in the culture supernatant was evaluated 48 h after the irradiation using enzyme-linked immunosorbent assay. Results Compared with the negative control, TGF-β1 in the culture supernatant decreased at the IPL irradiation of 10, 18, 27, and 36 J / cm2, whereas TGF-β1 increased at the IPL irradiation of 36 J/cm2× 2. In the IPL + inhibitor group, the concentration of TGF-β1 in the culture supernatant decreased compared with the controls (P<0.05). Conclusion IPL can suppress the secretion of TGF-β1 at the lower fluence and promote the secretion at a higher fluence. JNK inhibitor may play an inhibitive role when IPL regulates the TGF-β1 secretion in cultured human fibroblasts. IPL may stimulate TGF-β1 secretion of the fibroblast cells in human skin via JNK signal pathway.

Objective To investigate the optimal condition for in vitro cultivation of Trichomonas vaginalis for obtaining a better harvest of T\^vaginalis. Methods An isolate of T\^vaginalis from clinical specimens was cultivated in three different media with initial inoculation of 9\^0?10\+4/ml under pH 5\^6. Results There was distinct difference after 96h incubation in the cumulative harvest of T.vaginalis. The highest harvest was received in cysteine/liver/peptone/maltose medium, followed by the liver/peptone/maltose medium and soybean/liver/peptone/maltose medium. Conclusion The cysteine/liver/peptone/maltose medium may be a suitable environment for in vitro multiplication of T.vaginalis.

Objective To explore the energy feature of the spontaneous neural activity in depressive patients with diurnal rhythm.Methods Twenty depressive patients with diurnal rhythm,20 depressive patients without diurnal rhythm and 20 gender-,age-,education-matched healthy controls were scanned with 3.0 T MRI Scanner.According to Results of ANOVA,group-wise two sample t-test was completed.The correlation analysis were conducted between ALFF of significant difference brain regions and the score of Hamilton rating scale for depression.Results The brain regions showing significant differences among three groups were located in the right parahippocampal gyrus(MNI:18,-36,-9;K=136),left parietal lobe(MNI:-54,-45,42;K=86),left cerebellum crus(MNI:-84,-60,36;K=86) and right cerebellum crus(MNI:9,-78,-36;K=91)(P<0.05,Alphasim correct).Compared with non-diurnal rhythm group,the diurnal rhythm group showed significantly increased ALFF in the right parahippocampal gyrus.Compared with healthy group,the diurnal rhythm group showed significantly increased ALFF in right parahippocampal gyrus,left parietal lobe,and significantly decreased ALFF in left cerebellum crus 1 and right cerebellum crus 2.Compared with healthy group,the non-diurnal rhythm group showed significantly increased ALFF in left parietal lobe,and significantly decreased ALFF in left cerebellum crus 1 and bilateral cerebellum crus 2.No correlation between ALFF of significant difference brain regions and clinical symptoms was found(P>0.05).Conclusion The diurnal rhythm observed in depression may be associated with aberrant activity of parahippocampal gyrus.

Objective To perform a risk factor analysis of central retinal vein occlusion (CRVO) and branch retinal vein occlusion (BRVO),and compare the difference in risk factors between CRVO and BRVO.Methods Retrospective observational casecontrol study included 46 CRVO patients,33 BRVO patients and 79 control subjects with senile cataract or refractive error,the risk factors and blood lipid spectrum analysis were performed and compared.Results Multivariate linear regression analysis showed that higher serum levels of homocysteine (P ＜ 0.000 1),total cholesterol (P =0.003 0),lipoprotein (a) (P =0.027 0),hypertension (P =0.022 0) and shorter axial length (P ＜0.000 1) were significantly correlated with CRVO.BRVO was associated with higher serum levels of homocysteine (P ＜0.000 1),total cholesterol (P =0.008 0),hypertension (P =0.002 0),body mass index (P =0.004 0) and shorter axial length (P =0.001 0).There was no significant difference in risk factors between CRVO and BRVO patients on multivariate analysis.Conclusion Multiple systemic (hyperlipidemia,hypertension and hyperhomocystinemia) and ocular (shorter axial length) risk factors are associated with both CRVO and BRVO,but these risk factors are not different between CRVO and BRVO.

Objective To establish a method of measuring adipose content and fat distribution of the thigh in normal glucose tolerance (NGT) subjects,and to investigate its relation to insulin resistance.Methods Insulin sensitivity was evaluated by hyperinsulinemic and euglycemic clamp technique,and femoral adipose content and fat distribution were determined by MRI in 30 individuals with NGT including 15 with normal weight and 15 overweighted or obese subjects.Results Compared to normal weight group,the subscutaneous adipose tissue of thigh (SCAT) [(176.7?21.6) cm~2 vs (115.0?12.8 ) cm~2,P＜0.05],adipose tissue of thigh beneath the fascia (SFAT) [(75.4?4.4 ) cm~2 vs (57.5?4.7 ) cm~2,P＜0.01] and intermuscular adipose tissue (IMAT) [(28.3?3.2) cm~2 vs (14.5?1.1 ) cm~2,P＜0.01] were greater in overweight/obesity group.Overweight/ obesity group had lower insulin sensitivity( glucose disposal rate under steady state of hyperinsulinemic euglycemic clamp:4.54?0.43 vs 7.88?0.75,P＜0.01).SFAT and IMAT were significantly correlated with insulin sensitivity.SFAT showed the most marked correlation with insulin sensitivity.Multiple stepwise regression analysis showed that the increased SFAT played a pivotal role in insulin resistance.Conclusion The adipose content and fat distribution are highly correlated with insulin sensitivity and the adipose tissue of thigh beneath fascia may play the most significant role in insulin sensitivity.

OBJECTIVETo investigate the relationship between the second to the fourth digit ratio (2D:4D) and body mass index (BMI) in infertile men of the Han ethnic group in Ningxia.

METHODSUsing anthropometry, we calculated the mean ratio of 2D:4D and BMI of 197 infertile men and 148 normal healthy male controls, followed by analysis of their relationship.

RESULTSThe BMI was correlated positively with the 2D:4D ratio of the left hand in the infertile men (P < 0.05) and in the patients with a higher 2D:4D ratio of the left hand (P < 0.05), but negatively with the 2D:4D ratio of the righ/left (Dr-1) (left: P < 0.01; Dr-l: P < 0.05). The mean 2D: 4D ratio and BMI were both lower in the normal control than in the infertile men, with statistically significant differences in BMI (P < 0.05) and the 2D:4D ratio of the left hand (P < 0.05).

CONCLUSIONThere is a correlation between the 2D:4D ratio and BMI in infertile men.

Objective To investigate clinical characteristics and emergency managements of postrenal acute renal failure(ARV)induced by melamine in infant.Method Fluid therapy for urine alkalization and hydration,cistoscope drainage and peritoneal dialysis step by step were exerted in those who had both a history of certain milk intake and ARF according to the definition of pediatric ARF which developed by Pediatric Nephrology Assembly of Chinese Pediatric Association in 1994.Results Thirty-four postrenal ARF cases with anuria due to melamine in Nanjing Children's Hospital of Nanjing Medical University were involved in the study.Seventy cases(50%)re-ceived fluid therapy only.Nine cases(26.5%)received fluid thempy and eistoscope drainagemand 4 cases (11.8%)received fluid therapy and cistoscope drainage and peritoneal dialysis.Four cases(11.8%)received ur-gent peritoneal dialysis due to severe hyperkalemia.All cases(100%)survived.The urine pH at the first day.the second day,and after the second day in those who just pass away urine were 6.1±1.0、6.5±0.7.5.3±0.4,respectively(F=4.563,P=0.026).Conclusions Fluid therapy for urine alkalization and hydration and stop sequential thempy are effective in infant with postrenal ARF induced by melamine.

OBJECTIVE: To extract sperm DNA from mixed stain by the modified differential lysis method combined with silicon bead method and to evaluate its application value. METHODS: Fifty-two mixed stains containing female STR genotypes detected by differential lysis method were collected. The sperm DNA was extracted by the modified method combined with silicon bead method, then genotyped with the Identifiler Kit, and compared with the results of genotyping by the conventional differential lysis method as control. RESULTS: Of the 52 samples, 38 samples with sole male STR genotypes in all loci were detected. The detection rate of male STR genotypes was 98.08% through the modified method combined with silicon bead method. CONCLUSION The modified differential lysis method combined with silicon bead method can be used in extraction of sperm DNA from mixed stain.
DNA/isolation & purification* ; DNA Fingerprinting ; Female ; Genotype ; Humans ; Male ; Polymerase Chain Reaction ; Silicon ; Spermatozoa