?AIM:To investigate the measurement of central anterior chamber depth ( ACD ) in patients with acute primary angle - closure glaucoma ( APACG ) with A - scan ultrasound, Pentacam and ultrasonic biological microscope ( UBM) .?METHODS: Thirty-five patients (35 eyes) with APACG were selected, of whom central ACD were measured with A-scan ultrasound, Pentacam and UBM.?RESULTS: The measurement values of ACD with A-scan ultrasound, UBM and Pentacam were 1. 5633±0. 2089, 1. 5783 ± 0. 2067, 1. 6275 ± 0. 2296mm, which was equal variance tested by the homogeneity of variance, and was significant different by multiple comparision (F=4. 074, P=0. 026). The difference of ACD between the two groups of A-scan ultrasound and UBM, A-scan ultrasound and Pentacam, UBM and Pentacam were statistically significant ( P = 0. 032, 0. 023, 0. 012 ). Altman- Bland analysis showed that the three methods were not consistent with each other.?CONCLUSION: The ACD value of the APACG with the three methods is the largest using Pentacam, followed by UBM and A - scan ultrasound. In clinical the three methods with different advantages can complement each other, but cannot be replaced. In order to obtain more accurate results, we should combine the advantage and make comprehensive analysis.

Objective To study the correlation between storage time and the content of 5-hydroxymethylfurfural (5-HMF) in Corni Fructus because of the color change caused by storage time. Methods Corni Fructus samples of different storage time with 0, 1, 2, 3, and 4 years were collected. The contents of 5-HMF were determined by HPLC. Results The HPLC determination method of 5-HMF in Corni Fructus was established. The contents of 5-HMF varied from undetected value to 0.292 8%, with the increase of storage time of 0, 1, 2, 3, and 4 years and the color gradually deepened from red, dark red, reddish brown to brown. The contents of 5-HMF in black wine-prepared Corni Fructus were 0.954 4%-1.837%. Conclusion The browning of Corni Fructus is related to the production of 5-HMF. With the extension of storage time of Corni Fructus, the color gradually deepens and the content of 5-HMF increase significantly. The storage time of Corni Fructus can be suggested to be one year.

Objective To explore the plasma ghrelin levels in children and adolescents with short stature and the role of ghrelin in growth hormone-releasing hormone-growth hormone(GHRH-GH) axis.Methods One hundred and fifty-seven children(115 male,42 female) with short stature were selected.Fasting plasma sample was extracted from 10 mL vemous blood of the children with short stature.Insulin tolerance test and arginine stimulation test was performed initially to differential diagnosis.And blood samples was divided into 3 ca-tegories:37 cases of complete growth hormone deficiency (CGHD),52 cases of partial growth hormone deficiency(PGHD) and 68 cases of idiopathic short stature(ISS) during these two growth hormone(GH)provocative tests.Controls consisted of age and gender-match 20 health children.Plasma ghrelin levels were measured by radioimmunoassay.Serum GH was detected by chemiluminescence method,and serum insulin-like growth factor-1 (IGF-1) was measured by using enzyme linked immunosorbent assay.Fasting glucose,insulin,testosterone,estra-diol,luteinizing hormone,follicle-stimulating hormone were measured.Statistical analysis were conducted by using SPSS 11.5 software.Results The fasting ghrelin levels of CGHD group were significantly lower than that of ISS group and control group(Pa0.05).The ghrelin levels were positive correlated with the stimulated GH peak(r=0.176 P0.05).Conclusion Ghrelin has an important role on GH secretion and abnormal secretion of ghrelin might be a reason of growth hormone deficiency which due to hypothalamic abnormality.

Objective To analyze the characteristics of genotyping and gene polymorphism of Neisseria gonorrhoeae(N.go) with azithromycin(AZM)-resistance(AZM-R) and decreased susceptibility to ceftriaxone(CROD).Methods The minimum inhibitory concentration(MIC) of AZM and CRO were determined.AZM-R isolates were detected for mutations in 23S rRNA,mtrR and penA genes.Genotypes were analyzed by using N.go multi-antigen sequence typing(NG-MAST).Results All total of 485 isolates of N.go were detected.77(15.9%) strains were AZM-R(MIC≥1 mg/L),including 33(6.8%) isolates of AZM low-level resistant(AZM-LLR,MIC=1 mg/L) strains and 44(9.1%) isolates of AZM middle-level resistant(AZM-MLR,MIC≥2 mg/L) strains.There were more CROD(MIC≥0.125 mg/L) strains in AZM-MLR isolates(43.2%),compared with those in AZM-LLR isolates(18.2%,P<0.05).The detected rates of 23S rRNA,mtrR,penA single or combined mutations were without significant differences between AZM-LLR isolates and AZM-MLR isolates(P>0.05).Similar results were found between combined AZM-LLR/CROD isolates and combined AZM-MLR/CROD isolates(P>0.05).No mutation of A2059G and AZM high-level resistant(AZM-HLR,MIC≥256 mg/L) isolate were found.Among 77 AZM-R isolates,67 sequence types(ST) were identified by NG-MAST,of which 30 types were novel.Most ST were represented by a single isolate.Conclusion AZM-R and CROD isolates,presented in this area,might be deserved continuous surveillance to identify the mechanism of concurrent resistance.

Objective To investigate the relationship between V-raf murine sarcoma viral oncogene homolog B1 (BRAF)v600E mutation and radioactive iodine (RAI) uptake in distant metastases from papillary thyroid cancer(PTC).Methods From January 2011 to December 2012,40 PTC patients (21 males,19 females,average age 39.8 years) with distant metastases were recruited and divided into mutation group and wild group according to the BRAFv600E mutation in primary lesions.The clinical,pathological and serological differences were compared between the two groups.The relationship between BRAFv600E mutation and RAI uptake capability in distant metastases from PTC,as well as its relationship with Tg change after 131I treatment were investigated.Statistical analysis was performed with two-sample t test,x2 test or Fisher exact test.Results The BRAFv600E mutation rate was 30.0％ (12/40) in patients with metastases from PTC.There was no significant difference in clinical,pathological and serological features between mutation group (n =12) and wild group (n=28; t:from-0.533 to 1.728,x2:from-1.951 to 1.088,all P＞0.05).Twelve PTC patients had no RAI uptake in the distant metastases,of which 10 belonged to mutation group (83.3％,10/12) and 2 belonged to wild group (7.1％,2/28; x2=19.734,P＜0.05).BRAFv600E mutation group was more likely to have no RAI uptake in the distant metastases.Tg change after 131I treatment in 30 patients were analyzed.In the wild group,Tg level decreased in 66.7％ (14/21) patients,stabilized in 19.0％ (4/21)and increased in 14.3％ (3/21)patients.While there was no decrease of Tg in the mutation group (0/9).Two patients had increased Tg level and 7 patients (with no RAI uptake) kept stable in mutation group.Conclusions Due to poor RAI uptake capability in PTC patients with BRAFv600E mutation,both primary and metastatic sites may have poor response to 131I treatment.Molecular detection of BRAFv600E mutation might be helpful for choosing PTC with distant metastases and predicting the effect of 131 I treatment.

Objective To investigate the role of regulatory T (Treg) / T helper type 17 (Th17) cells in the pathogenesis of chronic spontaneous urticaria (CSU).Methods Eighty-nine patients with CSU were enrolled in this study,including 48 in active stage and 41 in remission stage.Forty-eight health check-up examinees,who were collected from the community hospitals in Guangzhou city,served as the healthy controls.Fluorescence-based realtime quantitative PCR was performed to determine the expression of transcription factors FOXP3 and RORγt in PBMCs from these subjects.Results Compared with the patients with CSU in remission stage and healthy controls,the patients in active stage showed a significantly higher level of FOXP3 mRNA (0.57 ± 0.19 vs.0.11 ± 0.21 and 0.13 ± 0.23,both P ＜ 0.05),but a significantly lower level of RORγt mRNA (0.43 ± 0.39 vs.0.89 ± 0.40 and 0.87 ± 0.43,both P ＜ 0.05).Conclusions The expression of Treg cell regulator FOXP3 increases,while the expression of Th17 cell regulator RORγt decreases in patients with CSU,suggesting that the imbalance between Treg and Th17 cells induced by the interaction between FOXP3 and RORγt may be involved in the pathogenesis of CSU.

OBJECTIVETo observe the neurological protective effects of progesterone (PROG) on focal cerebral ischemia/reperfusion injury in rats and to explore its possible mechanism.

METHODSOne handred and twenty male SD rats were divided into three groups randomly: sham-operated group, middle cerebral artery occlusion ( MCAO ) group and PROG + MCAO group( n = 40). The right temporary MCAO model was established by the line-embolism method. The PROG + MCAO group rats were according to 8 mg/kg intraperitoneal injection PROG, after that 30 min, the rats were suffered ischemia/reperfusion. After rats were suffered ischemia for 2 h and reperfusion 0, 24, 48, 72 h stress, the nervous functional defect degree were evaluated by longe scoring, and the expression of two-pore domain K channel 3 (TASK3) mRNA in brain tissue were detected by the real-time PCR.

RESULTSPROG (8 mg/kg) could significantly reduced the nervous functional defect degree in rats after ischemia/reperfusion 24, 48, 72 h (P < 0.05). The results of real-time PCR showed that the TASK3 mRNA expression in the brain tissue at all time points significantly decreased in MCAO group compared with sham-operated group (P < 0.05). However, compared with MCAO group, the expression of TASK3 mRNA in brain tissue at all time points dramatically increased in PROG + MCAO group (P < 0.05).

CONCLUSIONPROG can improve the nervous functional defect degree after focal cerebral ischemia/reperfusion injury in rats, and the mechanism might be associated with up-regulating the expression of TASK3 mRNA in brain tissue.

Animals ; Brain ; drug effects ; pathology ; Brain Ischemia ; drug therapy ; Infarction, Middle Cerebral Artery ; Male ; Neuroprotective Agents ; pharmacology ; Potassium Channels, Tandem Pore Domain ; metabolism ; Progesterone ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Reperfusion Injury ; drug therapy

Lanosterol synthase is encoded by the erg7 gene and catalyzes the cyclization of 2, 3-oxidosqualene, which is a rate-limiting step of the inherent mevalonate (MVA)/ergosterol metabolic pathway in Saccharomyces cerevisiae. The intermediate 2, 3-oxidosqualene is also the precursor of triterpenoids. Therefore, the cyclization of 2, 3-oxidosqualene is the key branch point of ergosterol and triterpenoids biosynthesis. Down-regulation of 2, 3-oxidosqualene metabolic flux to ergosterol in S. cerevisiae may redirect the metabolic flux toward the triterpenoid synthetic pathway reconstructed by the synthetic biology approach. To construct erg7 knockout cassette harboring the loxP-Marker-loxP element, long primers were designed, which were homologous to the sequences of both erg7 ORF and plasmid pUG66. The cassette was transformed into diploid wild strain INVSc1 by LiAc/SS Carrier DNA/PEG method and then erg7 gene haploid deficient mutant was obtained by homologous recombination. The results of semiquantitative PCR and real-time quantitative PCR revealed that erg7 expression level in erg7 gene haploid deficient mutant is one time lower than that in wild strain. The results of TLC and HPLC showed that the ergosterol content in deficient mutant decreased to 42% of that in wild strain.
Chromatography, High Pressure Liquid ; DNA Primers ; Down-Regulation ; Ergosterol ; metabolism ; Haploidy ; Intramolecular Transferases ; genetics ; Polymerase Chain Reaction ; Saccharomyces cerevisiae ; genetics ; Squalene ; analogs & derivatives ; metabolism

Pichia pastoris is one of the most important systems used in the field of molecular biology for the expression of recombinant proteins. The system has advantages of high expression, high stability, high secretion, easy high-density fermentation and low cost. Many factors affect the expression of recombinant protein, such as gene copy number, codon usage preference, type of promoter, molecular chaperones, glycosylation, signal peptide and fermentation process. In this review, research advances of the above aspects are summarized, which lay a foundation for improving the expression of recombinant proteins in P. pastoris.
Fermentation ; Gene Dosage ; Glycosylation ; Molecular Chaperones ; Pichia ; metabolism ; Promoter Regions, Genetic ; Protein Sorting Signals ; Recombinant Proteins ; biosynthesis

Objective To explore the changes of mTOR signaling in LPS/D-gal-induced acute hepatitis in mice . Methods Twenty-six healthy adult female ICR mice were divided into two groups:the control group and experimental group, 13 mice in each group .LPS/D-gal was used to induce acute hepatitis in the mice .The survival of mice was moni-tored within 24 hours after LPS/D-gal challenge .At 6 hours after challenge , samples of serum and liver tissue were collect-ed for further analysis.Results Injection of LPS/D-gal resulted in acute death of the mice within 24 hours.At 6 hours post LPS/D-gal injection , the blood levels of ALT and AST were significantly increased .The mRNA expression of inflammatory cytokines Tnfa and Il6 was up-regulated in LPS/D-gal-induced hapatitis , in which DNA fragmentation and activation of caspase-3 were subsequently observed .Immunoblot analysis showed that both mTOR pathway and NF-κB pathway were ac-tivated.Unexpectedly , inhibition of mTOR signaling could neither decrease the apoptosis in the liver nor increase the sur -vival of mice .Conclusions The results of the present study indicate that mTOR signaling may play pleiotropic roles in the pathogenesis of LPS/D-gal-induced hepatitis .