OBJECTIVETo explore the diagnostic value of the measurement of serum Golgi protein 73 (GP73) in the diagnosis of hepatocellular carcinoma (HCC).

METHODSEnzyme-linked immunosorbent assay (ELISA) was used to detect the serum GP73 in the 81 cases of HCC, 71 cases of chronic hepatitis or cirrhosis (CH/LC) and 65 cases of healthy blood donors, and to evaluate the sensitivity and specificity in the diagnosis of HCC through the ROC curves.

RESULTSThe average levels of serum GP73 in HCC, CH/LC and Normal groups were (152.67 +/- 33.59) ng/ml, (93.15 +/- 20.02) ng/ml and (58.95 +/- 17.29) ng/ml(o) After calculating through the ROC curves, 120 ng/ml was set as the optimal cut-off point, GP73 has a sensitivity of 77.80% and a specificity of 78.00%.

CONCLUSIONSGP73 as a serum marker in the diagnosis of HCC had a higher sensitivity than AFP, and the combined detection of GP73 and AFP could improve HCC diagnosis.

Carcinoma, Hepatocellular ; diagnosis ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Liver Cirrhosis ; blood ; Liver Neoplasms ; diagnosis ; Male ; Membrane Proteins ; blood ; alpha-Fetoproteins ; analysis

Objective To investigate influence factors on neonatal necrotizing enterocolitis (NEC) and find a preventive way against the said disorder. Methods A retrospective study was conducted of 63 cases NEC (observation group) and 70 cases non-NEC (control group) admitted to the neonatal intensive care units (NICU) of two hospitals from January 2010 to May 2014. The cases were grouped in 29 factors. The individual factors of 28 out of the 29 factors were analy-sised with SAS 9.1 statistical software, and the screened important factors were investigated by logistic regression. Re-sults The ratios of 4 factors, application of prenatal corticosteroids, breast feeding prior to the ailment, oral probiotics and hyperbilirubinemia in the observation group were lower than those in the control group, while the ratios of 14 factors were higher than those in the control group, premature rupture of membrane, asphyxia, respiratory distress syndrome, respiratory failure, umbilical venous catheters, oxygen uptake, sepsis, septic shock, disseminated intravascular coagula-tion (DIC), congenital heart disease, scleredema, hyponatremia, hypocalcaemia and transfusion. The differences of fac-tors mentioned were of statistic significance (P<0.05), while the differences of other factors were not statistically signifi-cant between the two groups (P>0.05). Logistic regression analysis indicated that breast feeding, oral probiotics and hy-perbilirubinemia were protective factors against NEC; while sepsis, congenital heart disease and transfusion were major factors that lead to NEC. Conclusion NEC resulted from a combination of multiple factors, with sepsis, congenital heart disease and transfusion as the leading ones. Breast feeding, oral probiotics, and non-excessive curing against hyper-bilirubinemia will help reduce the incidence of NEC.

OBJECTIVETo compare the sensitivities of MALDI-TOF MS and direct PCR sequencing on gene mutations detection of hepatitis B virus.

METHODS100 serum samples from chronic hepatitis B patients were collected, which consisted of 90 serum samples (study group) from 90 chronic hepatitis B patients received nucleoside analogues (NA) therapy for more than 1 year and HBV DNA titer still higher than 500 copies/ml and 10 serum samples (blank group) from 10 chronic hepatitis B patients never treated with antiviral therapy and HBV-DNA titer higher than 1 x 10(5) copies/ml. 9 known mutations associated with HBV P gene in these samples were detected by MALDI-TOF MS and direct PCR sequencing at the same time, TYPE4.0 software and Sequence Navigator software were used to analyze the results separately.

RESULTS(1) In study group, mutations were detected in 53 samples and the total mutation sites were 86 by MALDI-TOF MS with a positive detection rate of 58.89%, whereas only 19 samples were found with mutations and totally 28 mutation sites were detected by direct PCR sequencing, the positive detection rate was 21.11%. The positive detection rate by MALDI-TOF MS was higher than that by direct PCR sequencing and the difference was statistically significant (P < 0.05). In blank group, no mutations were detected by any method. (2) In study group, when the HBV DNA titers were at 500-1000 copies/ml, 10(3)-10(4) copies/ml and 10(4)-10(5) copies/ml, the positive mutation detection rates by MALDI-TOF MS were 50%, 52.08% and 77.27% respectively, higher than that by direct PCR sequencing, which were only 0%, 8.33% and 45.45%. The difference was still statistically significant (P < 0.05).

CONCLUSIONSMALDI-TOF MS had higher detection sensitivity for known mutation sites as compared to direct PCR sequencing method.

DNA Mutational Analysis ; Drug Resistance, Viral ; genetics ; Hepatitis B virus ; drug effects ; genetics ; Humans ; Mutation Rate ; Polymerase Chain Reaction ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

[Objective]To investigate the efficacy and safety of ALA-PDT following CO2laser preconditioning for recal-citrant plantar warts.[Methods]Patients with recalcitrant plantar warts were enrolled in this study,and received ALA-PDT treatment following CO2laser preconditioning. Cure rate and side effects were observed.[Results]Twenty patients were en-rolled and 85%(17/20)showed complete clearance of plantar warts after one to three times of ALA-PDT. PDT treatment time was once in two patients(10%),twice in five patients(25%)while three times in 10 patients(50%).No infection or scar tissue was observed.Five(25%)patients were infected with one type of HPV,while 15(75%)patients with two or mul-tiple types of HPV. No difference was observed in complete clearance rate between patients with single or multiple HPV gen-otypes infection.[Conclusions]Superpulse carbon dioxide laser pretreatment enhanced the efficacy of ALA-PDT treatment on recalcitrant plantar warts.Further study is needed to determine the association of HPV genotype with outcome of recalci-trant plantar warts.

This study is to investigate the expression of CyclinD1 in asthmatic rats and construct expression plasmids of sense and antisense CyclinD1 gene and transfect them to asthmatic airway smooth muscle cell to study the effects of CyclinD1 on the proliferation of airway smooth muscle cells in asthmatic rats. CyclinD1 cDNA was obtained by RT-PCR of total RNA extracted from the airway smooth muscle in asthmatic rats. The sequence was inserted into eukaryotic expression vector pcDNA3.1 (+) to recombinate the sense and antisense pcDNA3.1-CyclinD1 eukaryotic expression vector. The two recombinations and vector were then separately transfected into airway smooth muscle cell in asthmatic rats by using liposome. The expression level of CyclinD1 was certificated by Western blotting analysis. The proliferations of ASMCs isolated from asthmatic rats were examined with cell cycle analysis, MTT colorimetric assay and proliferating cell nuclear antigen (PCNA) immunocytochemical staining. Results showed (1) Compared with control group, the content of CyclinD1 was significantly increased; (2) It was comformed by restriction endonucleasa digestion and DNA sequence analysis that the expression plasmid of sense and antisense CyclinD1 were successfully recombinated. There was significant change of CyclinD1 expression between vector and sense CyclinD1 transfected cells, and the expression level of CyclinD1 in ASMC transfected with antisense CyclinD1 was lower than that in vector transfected cells (P <0.01); (3) In the asthmatic groups, compared with the vecter group, the percentage of S + G2M phase, absorbance A value of MTT and the expression rate of PCNA protein in ASMC transfected with pcDNA3. 1-CyclinD1 vector significantly increased. The values decreased remarkably in the pcDNA3,1-as CyclinD1 group. Statistical analysis revealed that there were significant differences in these indicators of cell proliferation in three groups (P <0.01). In the normal groups, statistical analysis revealed that there were significant differences in the percentage of S + G2M phase, a value of MTT and the expression rate of PCNA protein in three groups (P <0.01). Sense CyclinD1 eukaryotic expression vectors could have a positive effect on the proliferation of ASMC, however the antisence one have a negative effect, which implicated that CyclinD1 might contribute to the process of airway smooth muscle cell proliferation.
Animals ; Asthma ; pathology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Codon ; genetics ; pharmacology ; Cyclin D1 ; agonists ; antagonists & inhibitors ; genetics ; DNA, Antisense ; genetics ; pharmacology ; Disease Models, Animal ; Gene Expression ; Genetic Vectors ; genetics ; Male ; Myocytes, Smooth Muscle ; drug effects ; pathology ; Rats ; Rats, Sprague-Dawley ; Recombination, Genetic ; genetics ; Respiratory System ; Reverse Transcriptase Polymerase Chain Reaction ; Transduction, Genetic ; Transfection

OBJECTIVETo study the effects of different parameters (frequency, intensity, needle-retained time and treatment interval) of electroacupuncture at Fenglong (ST 40) for adjusting blood lipids, so as to find out the optimization parameter.

METHODSFifty-four cases meeting the criteria for hyperlipoidemia were randomly divided into 27 groups with orthogonal design L27 (3(13) ). According to the orthogonal design program they were treated with electroacupuncture at Fenglong (ST 40). Ten sessions constituted one course with a one week's interval between two course. The treatment was given for 2 courses.

RESULTS(1) The parameters of EA at Fenglong (ST 40) for regulating blood lipids in primary and secondary orders are: frequency, needle-retained time, interval of treatment, intensity. (2) The parameters of EA at Fenglong (ST 40) for various programs in regulating various blood lipids are: for TG, frequency AM 50 Hz, needle-retained time 20 ain, intensity 1 mA, twice each week; for TC, frequency AM 100 Hz, needle-retained time 30 min, intensity 1 mA, once every other day; for LDL-C, frequency Am 100 Hz, needle-retained time 30 min, intensity tolerable and comfortable, once every other day.

Acupuncture Points ; Aged ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Electroacupuncture ; methods ; Female ; Humans ; Hyperlipidemias ; blood ; therapy ; Lipids ; blood ; Male ; Middle Aged

BACKGROUNDAirway smooth muscle (ASM) is suspected to be a determining factor in the structural change of asthma. However, the role of protein kinase C alpha (PKCalpha) and cyclin D1 involved in the dysfunction of ASM leading to asthmatic symptoms is not clear. In this study, the central role of PKCalpha and cyclin D1 in ASM proliferation in asthmatic rats was explored.

METHODSThirty-six pathogen-free male Brown Norway (BN) rats were randomly divided into 2 groups: control groups (group N1, N2 and N3) and asthmatic groups (group A1, A2, and A3). Groups A1, A2 and A3 were challenged with ovalbumin (OA) for 2 weeks, 4 weeks and 8 weeks respectively. Control animals were exposed to an aerosolized sterile phosphate buffered saline (PBS). The ASM mass and nucleus numbers were studied to estimate the degree of airway remodeling by the hematoxylin-eosin staining method. PKCalpha and cyclin D1 expression in the ASM cells was detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. The relation between PKCalpha and cyclin D1 was assessed by linear regression analysis. PKC agonist phorbol 12-myristate 13-acetate (PMA), PKC inhibitor Ro31-8220 and an antisense oligonucleotide against cyclin D1 (ASOND) were used to treat ASM cells (ASMCs) obtained from the 2 weeks asthmatic rats. The cyclin D1 protein expression level was detected by Western blotting.

RESULTSCompared with the control group, the PKCalpha and cyclin D1 mRNA levels were increased in the asthmatic group. Similar to RT-PCR results, immunohistochemistry analysis for PKCalpha and cyclin D1 expression revealed an increased production in ASMCs after allergen treatment for 2, 4 and 8 weeks compared with the respective control groups. No difference in expression of PKCalpha and cyclin D1 in ASM were found in the 2, 4 or 8 weeks asthmatic rats. There were significant positive correlations between PKCalpha and cyclin D1 expression, both transcriptionally (r = 0.944, P < 0.01) and translationally (r = 0.826, P < 0.01), in ASM. The content of cyclin D1 in asthmatic ASMCs increased after being stimulated by PMA, and decreased when induced by Ro31-8220. ASOND targeting for cyclin D1 lowered the expression of cyclin D1 induced by PMA.

CONCLUSIONSIncreased expression of PKCalpha and cyclin D1 in ASM along with smooth muscle structure changes might implicate PKCalpha and cyclin D1 participation in the proliferation of ASM and contribute to the pathogenesis of asthma after repeated allergen exposure in rats. The results suggested that cyclin D1 might be downstream of PKC signal transduction pathway.

Animals ; Asthma ; pathology ; Cell Proliferation ; Cyclin D1 ; genetics ; physiology ; Lung ; pathology ; Male ; Myocytes, Smooth Muscle ; pathology ; Protein Kinase C-alpha ; genetics ; physiology ; RNA, Messenger ; analysis ; Rats ; Rats, Inbred BN

We preliminarily investigated the role of NLRP3 inflammasome in Helicobacter pylori (H.pylori) Tipa-induced pro-inflammatory cytokines secretion in PMA-differentiated human acute monocytic leukemia cell line THP-1.PMA-differentiated THP-1 cells were treated with pure recombinant Tips protein.The secretion levels of TNF-α,IL-1β and IL-18 in supernatant of culture medium were detected by ELISA.Then we blocked MyD88/NF-κB and NLRP3/Caspase-1 pathways by PDTC or the general ROS scavenger,NAC,respectively,and determined the secretion levels of proinflammatory cytokines and the expression levels of NLRP3 and Caspase-1.The results showed that Tips protein can significantly induced the secretion of TNF-α,IL-1β and IL-18 in THP-1 cells in a time and dose-dependent manner.Levels of TNF-α,IL-1β and IL-18 approached their peaks at 6 h post-treatment by 40 μg/mL of Tipα protein (P＜0.05).Moreover,the blockade of NF-κB signaling pathway by PDTC can inhibit the secretion of proinflammatory cytokines and the expression of NLRP3 and Caspase-1.When THP-1 cells were pre-treated with ROS scavenger NAC,the Tipα-induced increased IL-1β and IL-18 secretion was obviously eliminated (P＜0.05),while TNF-α level had no significant difference,and the expression levels of Caspase-1 and NLRP3 also have a significant decrease.Our results demonstrated that Tipα can promote THP-1-drived macrophages to secrete proinflammatory cytokines TNF-α,IL-1β and IL-18,and the NLRP3/Caspase-1 pathway may be involved in the Tipα protein-induced IL-1β and IL-18 secretion.

OBJECTIVETo investigate the clinical effects of electroacupuncture (EA) at Fenglong (ST 40) on blood lipids.

METHODSTwo hundred and four patients of hyperlipidemia were randomly divided into a Fenglong group and a Xuezhikang group, 102 cases in each group. The patients in the Fenglong group were treated with electroacupuncture at Fenglong (ST 40). After arrival of qi, the needles were connected with acupoint nerve stimulator (LH 202 H type, HANS). The primary parameters of EA: for high triglycerides (TG) type, AM 50 Hz, intensity 1 mA, needle-retained time 20 min, twice per week; for high cholesterol (CHO) type, AM 100 Hz, intensity 1 mA, needle-retained time 30 min, thrice per week; for high low-density-lipoprotein (LDL-C) type, the same parameters as the high CHO type except the tolerable and comfortable intensity; for the mixing type, corresponding methods were alternatively used. The patients in the Xuezhikang group received Xuezhikang capsule orally, 2 capsules each time and twice daily, for total 11 weeks.

RESULTSThe total effective rates of the Fenglong group and the Xuezhi-kang group were 83.0% and 85.9%, respectively, with no significant difference between the two groups (P > 0.05), and there was no significant differences in the function of regulating blood lipids between the two groups (all P > 0.05). After one month follow-up survey, the total CHO, TG and LDL-C decreased and high-density-lipoprotein (HDL-C) increased, of which there was a significant difference in TG reduction (P < 0.05). There were no relapses in both groups.

CONCLUSIONEA at Fenglong (ST 40) can effectively regulate blood lipids with a better after-effect, which can be applied as a safe and effective method to replace medication for regulating blood lipids.

Acupuncture Points ; Adult ; Aged ; Cholesterol ; blood ; Electroacupuncture ; Female ; Follow-Up Studies ; Humans ; Hyperlipidemias ; blood ; therapy ; Lipids ; blood ; Male ; Middle Aged ; Triglycerides ; blood

Objective To investigate the life quality status in autistic disorder children and its influencing factors.Methods Pediatric Quality of Life Inventory (PedsQL4.0) was used to measure the life quality of 200 children with autistic disorder and 120 healthy children.Application of Gesell Development Diagnosis Scale (1981) test was used for intelligence development.Clancy Autism Behavior Scale,Autism Behavior Checklist,and Childhood Autism Rating Scale were used to evaluate the illness degree.Results The life quality of autistic disorder group was lower than that of control group in the scores of physical functioning,emotional functioning,social functioning,mental domain and the totals cores of PedsQL,the differences were significant (all P ＜0.01).High-functioning autism in all aspects and the overall life quality were higher than those of low function group,the differences were statistically significant (all P ＜0.01).By using variance analysis,intelligence factors had an impact on the patient's life quality,the difference was statistically significant(P ＜ 0.01).Correlation analysis results suggested that the autism condition degree was heavier,the intelligence level was lower,the life quality was worse,and the difference was statistically significant (P ＜ 0.05).Conclusion Children with autistic disorder suffer grievous influence in life quality.