OBJECTIVETo explore the effect of Lignum Sappan (LS) containing serum on the proliferation cycle arrest of human lung cancer cell line PG and its molecular mechanism.

METHODSThe lung cancer PG cells were divided into four groups, i.e., the blank control group, the LS group, the LS plus cisplatin group, and the cisplatin group. They were cultured by RPMI-1640 with 20% blank serum, RPMI-1640 with 20% LS containing serum, RPMI-1640 with 20% LS containing serum plus 1 microg/mL cisplatin, and RPMI-1640 with 20% blank serum plus 1 microg/mL cisplatin, respectively. The morphology of PG cells was observed using light microscope and laser scanning confocal microscope in each group. The cell cycle arrest was observed using flow cytometry. The expression of P16 and Rb1 mRNA was tested by PCR method.

RESULTSUnder the light microscope and laser scanning confocal microscope, the apoptosis degree of PG cells in the LS group was significant, but less than that of the LS plus cisplatin group as well as the cisplatin group. Compared with the blank control group, the proportion of PG cells increased at G0/ G1 and S phases (P < 0.05) and decreased at G2/M phase (P < 0.01) in the LS group; The proportion of PG cells increased at G2/M and S phases (P < 0.05, P < 0.01) and decreased at G0/G1 phase (P < 0.01) in the LS plus cisplatin group as well as the cisplatin group. Compared with the LS group, the proportion of PG cells increased at G2/M and S phases (P < 0.05, P < 0.01) and decreased at G0/G1 phase (P < 0.01) in the LS plus cisplatin group as well as the cisplatin group. There was no statistical difference in PG cells at each phase between the cisplatin group and the LS plus cisplatin group (P > 0.05). The expression of P16 and Rb1 mRNA increased in the LS group, when compared with the blank control group. They also increased in the cisplatin group and the LS plus cisplatin group, higher than that of the LS group (P < 0.05). There was no statistical difference in the expression of P16 and Rb1 mRNA between the cisplatin group and the LS plus cisplatin group (P > 0.05).

CONCLUSIONLS containing serum induced PG cell apoptosis by up-regulating the mRNA transcription levels of P16 and Rb1, thus resulting in PG cell arrest at G0/G1 and S phases, which was different from the manner of cisplatin (achieved by arresting PG cells at G2/M and S phases through regulating cyclinB1 mRNA transcription).

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Lung Neoplasms ; pathology

Objective To study the normal values of blood pressure in healthy full-term infants in the first 7 days of life,and to determine the critical hypertension and hypotension in newborn infants.Methods Omni-Trak~(TM)NVS monitor with DINAMAP was used to measure the blood pressure.Systolic,diastolic and mean blood pressure were measured every day by DLNAMAP in a cohort of 50 healthy full-term infants in the first 7 days of life.Results 1.Systolic,diastolic and mean blood pressure of healthy full-term infants were significantly influenced with the age of birth.2.Stepwise linear multiple regressions was used to examine the multiple correlations among days of life,birth weight and gestational age.3.Optimal regression equations were built respectively.SBP=32.4+0.7X_1+(3.7X_2+)(0.6)X_3(mm Hg).DBP=13.1+ 0.6X_1+2.9X_2+0.6X_3(mm Hg).MAP=17.3+0.4X_1+2.4X_2+0.8X_3(mm Hg).(3.MAP=)(DBP+)(0.45)(SBP-DBP)(mm Hg).4.The critical hypertension and hypotension in newborn infants were obtained.Conclusions (Du)ring the first 7 days of life,there is a progressive rise in blood pressure of healthy full-term infants.There are linear reliance correlations between systolic,diastolic,mean blood pressure and day of life,birth weight gestational age.The critical hypertension and hypotension in newborn infants is important for clinic diagnosis.

Objective To investigate the characteristic of ?-amyloid precursor protein (A?) processing in activated platelet in AD.Methods Thirty-six sporadic AD patients and 30 control subjects were included in this study.Blood was collected from the subjects to separate platelets.After treated by thrombin,the soluble amyloid precursor protein (APP) level in the snpernatants of platelets from 36 were analyzed by means of western blot with a specific antibody recognizing soluble APP.Meanwhile A? level was measured by radioimmunoassay.Results After treated with thrombin,the level of soluble APP in the supernatants of platelets in patients with AD decreased by 31.0% (P

Enterotoxins ; analysis ; Humans ; Microbial Sensitivity Tests ; Staphylococcal Food Poisoning ; microbiology ; Staphylococcal Protein A ; analysis ; Staphylococcus aureus ; isolation & purification

OBJECTIVETo study the effects of sodium butyrate (SB) on growth, apoptosis and telomerase activity in Hep-2 cells.

METHODSGrowth inhibition effect of SB on Hep-2 cells was assessed by methyl thiazolyl tetrazolium (MTT) assay. Morphological alterations were observed by electronic microscope. Cell apoptosis was confirmed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling (TUNEL) method, DNA fragmentation and flow cytometry (FCM). Cell cycle was analyzed by FCM. Telomerase activity was examined by telomeric repeat amplification protocol (TRAP)-silver staining. The expression status of telomerase subunits was analyzed by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSA time-and dose-dependent inhibition was detected in cells treated with SB. Typical morphological changes of apoptotic cells were observed under electronic microscopy. The characteristic DNA fragmentation of apoptotic cells was detected by agarose gel electrophoresis. Apoptosis and the changes of cell cycle were confirmed by TUNEL method and FCM. The apoptosis indexes of the cells before treatment and at 72 h after SB (2.5 mmol/L) treatment were 2.27 +/- 1.18 and 33.50 +/- 2.75 respectively, the apoptosis rates were 2. 86% and 31. 28% respectively, the proportion of the cells at G0/G1 stage were 50.38% and 70.88% respectively, the proportion of the cells at S stage were 27.40% and 8.20% respectively, and the proliferation indexes of the cells were 49.62% and 29.12% respectively. Telomerase activity and expression level of human telomerase reverse transcriptase (hTERT), the key subunit of telomerase, decreased after SB treatment. No significant changes were observed in the expression of human telomerase RNA (hTR) and human telomerase associated protein (hTP1), the other two subunit of telomerase.

CONCLUSIONSB could inhibit growth of Hep-2 cells and induce apoptosis in the cells, and inhibit telomerase activity by decrease expression level of hTERT.

Apoptosis ; drug effects ; Butyrates ; pharmacology ; Cell Cycle ; drug effects ; Hep G2 Cells ; Humans ; Sodium ; pharmacology ; Telomerase ; metabolism

OBJECTIVE To explore the influences of Qingre Chushi Jiedu Formula on skin physiological indexes and im-mune indexes in patients with acne due to damp-heat in spleen and stomach.METHODS 102 cases with acne caused by damp-ness-heat in the spleen and stomach hospitalized in our hospital from June,2015 to September,2016 were chosen and randomly divided into the observation group and control group based on random number method,with 51 cases in each group.Patients in the control group were given Danshentong Capsule,while those in the observation group were treated with Qingre Chushi Jiedu Formula.The treatment lasted for four weeks in total.Then the improvement of clinical symptoms were observed,with skin physiological indexes(including skin moisture content,skin oil and pH value),serum interleukin(IL-2,IL-4,IL-10)as well as interferon(INF-γ) being tested before and after treatment to evaluate the clinical therapeutic effects of both groups.RESULTS Scores of lesion extent,lesion amount,lesion color,lesion swelling and pain as well as total lesion scores and TCM symptom scores of both groups went down significantly(P<0.01),with the observation group being better than the control group(P<0.01).The total therapeutic effect rate of observation group was 90.2%,notably higher than 66.7% of the control group(P<0.01);Heart,liver and kidney function,together with blood routine test and urine routine test of the both groups were within the normal range,with no side effects being noticed.The skin moisture extent of the two groups after treatment went up evi-dently(P<0.01),whereas the skin oil went down significantly(P<0.01).The two groups enjoyed evidently increased level of IL-2 and INF-γafter treatment(P<0.01)and decreased level of IL-4 and IL-10(P<0.01).while the recurrence rate of the observation group either four weeks or eight weeks after treatment was much lower than that of the control group(P<0.05). CONCLUSION Qingre Chushi Jiedu Formula can efficiently improve the clinical symptoms of patients with acne due to damp-heat accumulation in spleen and stomach and reduce the recurrence rate,whose mechanism might be related to the improve-ment of patients'skin physiological indexes and immune function.

OBJECTIVETo investigate the association between a new polymorphism (IVS3-20 T>C GenBank accession number: AY463003) in intro 3 of the parkin gene and the risk for Parkinson's disease (PD) in Chinese, particularly the relation between this polymorphism and the age of onset of PD patients.

METHODSPD was diagnosed according to the criteria of Core Assessment Program for Intracerebral Transplantations(CAPIT). All patients and controls were examined by two neurologists and were of the Han ethnic background. Polymerase chain reaction (PCR), denaturing high performance liquid chromatography (DHPLC) and sequencing were used to determine the genotype of each subject.

RESULTSA total of 312 PD patients (including 99 early-onset PD patients and 213 late-onset PD patients) and 236 controls were studied. The C/C homozygote was not found in this study. Chi-square analysis revealed that the frequencies of the C allele and T/C genotype were higher in total PD group but were not statistically different from those of the control group (P=0.6350 and 0.6331, respectively). After being stratified by age of onset, the frequency of T/C genotype was significantly higher (OR=3.52, 95%CI 0.97-13.13) in PD group with an onset age at or below 45 years old (7.07%), compared with that in the control group (2.12%). Similarly, C allele was much higher (OR=3.42, 95%CI 0.96-12.57, P=0.0276) in the early-onset PD group (3.90%) than that in the control group (1.06%). The linear trend analysis showed that both the T/C genotype and C allele increased significantly in the PD group with the increase of the onset age [chi-square(trend of Genotypes)=4.414, P=0.036; chi-square(trend of Alleles)=4.344, P=0.037]. On the other hand, there was no difference in the frequencies of allele and genotype between the late-onset PD patients and controls.

CONCLUSIONThe above results suggest that the parkin IVS3-20 T>C polymorphism might be a genetic risk factor for early-onset PD in Chinese.

Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Base Sequence ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Parkinson Disease ; genetics ; Polymorphism, Genetic ; Sex Distribution ; Ubiquitin-Protein Ligases ; genetics

OBJECTIVETo investigate the possible association of IVS5-5G>A polymorphism, positioned in the upstream region of exon 5 of PINK1 gene with the risk for sporadic late onset Parkinson disease (LOPD) in Chinese.

METHODSIntronic regulatory sequence analysis was performed using the web-based in-silico analysis. The authors performed an association study using a case-control series (comprising 382 LOPD patients and 336 controls, Chinese of Han ancestry). Genotyping was performed by PCR-based denaturing high performance liquid chromatography (DHPLC) combined with sequencing analyses. Allele and genotype frequencies were compared by the Chi-square test.

RESULTSIn-silico analysis showed that the intronic IVS5-5G>A polymorphism was located within acceptor site of exon 5 and may be the functional single polymorphism (SNP) in the regulatory region with impact on the splicing of PINK1 gene. Those result yielded statistical significant evidence for the association of PINK1 IVS5-5G>A polymorphism with risk for typical PD in Chinese Han population (OR=1.95, 95%CI: 1.29-2.94, P=0.0012). Homozygote of A allele may have increased risk for LOPD (OR=2.45, 95%CI: 1.27-4.72, P=0.009).

CONCLUSIONThe authors provide the first evidence that the common genetic variation PINK1 IVS5-5G>A may contribute to the risk of LOPD in Chinese population.

Age Distribution ; Age of Onset ; Aged ; Aged, 80 and over ; Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Exons ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Parkinson Disease ; genetics ; Polymorphism, Genetic ; Protein Kinases ; genetics ; Sex Distribution

OBJECTIVETo screen for polymorphisms in alpha 4 subunit (principal subunit of nAChR) gene (CHRNA4).

METHODSDNA was extracted from leukocytes of all subjects including 100 healthy senior people and 100 patients with Parkinson's disease (PD). The exons and adjacent intron regions of CHRNA4 were amplified with PCR. SNPs were screened by denatured high performance liquid chromatography (DHPLC) techniques and restriction fragment length polymorphisms. Potential mutations were confirmed by sequencing.

RESULTSTotal 10 polymorphisms were detected and identified in coding and adjacent intron regions of nAChR alpha 4 gene, that are 420C/T (0.873/0.127), 870C/T (0.828/0.172), 1440A/C (0.858/0.142), 1860C/T (0.738/0.262), 1890C/T (0.605/0.395), intron 5 +14T/C (0.553/0.447), intron 2 +22G/A (0.873/0.127), intron 3 +182 Del22bp (0.813/0.187), 1758C/T and 1809C/T (reference for coding sequence is GenBank SNPs 000744), of which the last three are novel mutations. PD patients appeared higher frequency of deletion in intron 3+182(0.235) than normal controls (0.140)(P=0.015).

CONCLUSIONnAChR alpha 4 gene is polymorphic. PD patients take higher frequency of intron3+182 Del 22 bp.

Aged ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Female ; Humans ; Male ; Molecular Sequence Data ; Parkinson Disease ; genetics ; Polymorphism, Genetic ; Receptors, Nicotinic ; genetics ; Sequence Analysis, DNA

OBJECTIVETo investigate the association between the C609T polymorphism of NADP (H): quinoneoxidoreductase 1 (NQ01) gene and decreased cognitive function and sporadic Alzheimer's disease (AD) in a community cohort.

METHODSPolymerase chain reaction (PCR), denaturing high performance liquid chromatography (DHPLC) and sequencing were used to determine the genotype of NQ01 in 110 subjects without cognitive dysfunction, 21 with cognitive dysfunction, and 65 AD patients from a community cohort.

RESULTSSignificantly different distributions of C/T and T/T genotypes were found between MMSE normal and abnormal subjects (OR=2.8, 95%CI 0.96-8.18, P=0.024), and between AD patients and healthy controls (OR=3.27, 95% CI 1.54-6.94, P=0.001), respectively. The frequencies of T allele of NQ01 C609T were significantly higher in MMSE abnormal subjects and AD patients (P=0.034 and 0.005) as compared to normal controls.

CONCLUSIONThe C609T polymorphism of NQ01 gene may be a genetic risk factor for cognitive dysfunction and sporadic AD in Chinese population.

Alzheimer Disease ; genetics ; Base Sequence ; Cognition Disorders ; genetics ; Molecular Sequence Data ; Mutation ; NAD(P)H Dehydrogenase (Quinone) ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Risk Factors ; Sequence Analysis, DNA