Anemia, Refractory ; Humans ; Male ; Middle Aged ; Thrombocytosis

Proteolytic degradation has been a severe problem when Pichia pastoris is employed to express recombinant proteins.One alternative method to circumvent this problem is to construct protease gene disruptant.However,the main study of gene disruption is focused on nonrecombinant Pichia pastoris rather than recombinant strain.In our study,we established two different methods to directly disrupt PRC1 and KEX1 gene in recombinant Pichia pastoris.On the basis of this,we further discussed and compared the application and advantages of both methods.

OBJECTIVE: To explore the value of estimating chronologic age based on the grades of mandibular third molar development. To evaluate whether mandibular third molar could be used as an indicator for estimating the age under or over 18 years. METHODS: The mineralization status of mandibular third molar of 1 845 individuals aged 10 - 30 was graded and marked based on Demirjian's classification of grades reformed by Orhan. Gender difference was examined by t-test. A cubic regression model was established to analyze the correlation between third molar and chronologic age. Each grade of age cumulative distribution diagram and ROC curve was respectively performed to evaluate the relationship between third molar and the age of 18. Using Bayes discriminant analysis, an equation was established for estimating the age of 18. RESULTS: The inner-rater reliability was 0.903. Statistical analysis showed a moderate correlation between age and grade. Significant differences of both genders were found only in grade D and H (P < 0.05). Males at the grades from 1 to D and females at the grades from 1 to C were under 18 years old, and both males and females at grade H were over 18 years old. The area under the ROC curve was 0.797 (P < 0.05). CONCLUSION Third molar development shows a high correlation with age, and combined with other indicators, it can be used to estimate the age of 18.
Age Determination by Teeth/methods* ; Asian People ; Bayes Theorem ; China ; Female ; Forensic Dentistry ; Humans ; Male ; Molar, Third/diagnostic imaging* ; Radiography, Panoramic ; Reproducibility of Results ; Sex Characteristics

OBJECTIVETo record the electrical activities of Antirior cingulate cortex (ACC) neurons by in vivo multi-channel recording methods using the model of complete freund's adjuvant (CFA) induced conditioned place avoidance (C-CPA), which has been set up in our previous studies.

METHODSThe electrode was self-made and the CPA responses were recorded by in vivo multi-channel recording method.

RESULTS(1) The electrical activities of ACC neurons could be successfully recorded by the self-made electrode. (2) Before or after the injection of CFA, rats were respectively conditioned to the different place. The firing rates of ACC neurons in the CFA-paired place vs that in the non-CFA-paired place was (0.853 ± 1.377) imp/s vs (0.221 ± 0.971) imp/s (P < 0.05, n = 26). (3) The CPA responses in the CFA-paired place vs that in the non-CFA-paired place were (303.55 ± 61.77)s vs (140.32 ± 33.52)s(P < 0.05, n = 6).

CONCLUSIONThe firing rates of rACC (rostral Anterior Cingulate Cortex) neurons were involved in the occurrence of the affective pain.

Animals ; Electrodes ; Freund's Adjuvant ; Gyrus Cinguli ; cytology ; Neurons ; cytology ; Pain ; diagnosis ; Pain Measurement ; methods ; Rats ; Rats, Sprague-Dawley

Japan moxibustion methods originate from China and form moxibustion methods with Japanese characteristics and diathermal moxibustion as main, in combination with Japanese practice in development course. In the present paper, diathermal moxibustion and folk traditional moxibustion methods in Japan are introduced and moxibustion methods between China and Japan are preliminarily compared.
Humans ; Japan ; Medicine, East Asian Traditional ; Moxibustion ; methods

Objective Platelet activating factor(PAF),which has been implicated in the pathophysiology of inflammation in asthma,is degraded and inactivated by PAF acetlhydrolase(PAF AH).To investigate the association of PAF AH activity with genotype in asthmatic children.Methods We studied 57 asthmatic children and 30 normal controls. The plasma PAF AH genotype was detected as representative case with 3 different genotypes (Val/Val,Val/Phe and Phe/Phe) by allele specific polymerase chain reaction(AS PCR).The PAF AH activity in plasam was examined by the changes of substrate assay.Results In severe asthmatic individuals plasma PAF AH activities were lower than those of mild or moderate groups and control group,and plasma PAF AH activition was absent 15.4 %.In another three groups plasma PAF AH activation were absent 2 %-3 %.There was significant difference of plasma PAF AH activity among 3 groups of genotype(Val/Val,Val/Phe and Phe/Phe).In the similar genotype, there was no significant difference of plasma PAF AH activity between the groups of control and asthma.Conclusions There was imbalace of PAF/PAF AH in asthmatic children. In severe asthmatic individuals plasma PAF AH activities were lower than those of mild or moderate groups and control group. PAF AH(Val279Phe) gene mutation was related with plasma PAF AH activity.

Objective:To compare the dosimetric difference between knowledge-based planning (KBP) volumetric modulated arc therapy (VMAT) and intensity-modulated radiotherapy (IMRT) models for predicting the dose distribution during IMRT, aiming to investigate the feasibility of VMAT model to predict the IMRT plans.Methods:Fifty prostate cancer patients who had completed radiotherapy were selected. Manual planning was performed on each selected patient to generate the corresponding IMRT and VMAT plans. The IMRT and VMAT manual plans of the 40 randomly-selected patients were adopted to generate the KBP VMAT and IMRT models. The remaining 10 patients were utilized to predict IMRT plans. VMAT library-derived IMRT model (V-IMRT) and IMRT library-derived IMRT model (I-IMRT) were generated. Dosimetric parameters related to organ-at-risks (OARs) and planning target volume (PTV) were statistically compared among the manual IMRT (mIMRT), V-IMRT and I-IMRT plans.Results:Compared with the mIMRT plan, I-IMRT could significantly better control D max of the PTV ( P=0.039), whereas V-IMRT and I-IMRT plans could better protect the bladder and bilateral femoral heads (both P<0.05). V-IMRT plan could better protect the D max of bilateral femoral heads and the D 15% of the right femoral head (both P<0.05), whereas no significant difference was observed in other OARs and PTV (all P>0.05). Conclusions:Compared with the manual plans, KBP IMRT plan has significant advantages in protecting the OARs. KBP VMAT and IMRT models are both feasible in clinical practice, which yield equivalent accuracy for predicting IMRT plan.

OBJECTIVE: To perform the validation and analysis of forensic parameters of Goldeneye DNA ID 26Y system. METHODS: Based on the validation rules of Scientific Working Group on DNA Analysis Methods (SWGDAM), the kit was assessed from several parts, as test of PCR system, reproducibility, accuracy, and sensitivity, etc. And Y-STR loci of 517 unrelated healthy individuals from Eastern China were genotypes by this kit. The distribution and frequency of haplotype were calculated and forensic parameters of the kit were assessed. RESULTS: The complete profiles can be obtained even when the PCR reaction volume with 6.25 microL. And correct profile was obtained with DNA down to 125 pg. No reproducible peaks were detected with the DNA of common animals and microorganism with the kit. For the male-male mixture testing, average 70% of the minor alleles were obtained when the ratios of 1:19 and 19:1. For the male-female mixture testing, results showed that the sensitivity of the kit was no compromised with the addition of female samples. CONCLUSION The validation studies demonstrated that Goldeneye DNA ID 26Y system has good sensitivity and specificity, and suitable for mixture testing. The polymorphism of 26 Y-STR loci included in this kit are good for forensic application.
Alleles ; Animals ; Asian People/genetics* ; China ; Chromosomes, Human, Y ; DNA ; DNA Fingerprinting/standards* ; Female ; Forensic Genetics/methods* ; Genotype ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVE: To investigate the genetic data of 21 autosomal STR included in Goldeneye™ DNA ID 22NC Kit in Chinese Han nationality and to evaluate the forensic application. METHODS: By detected 500 unrelated healthy individuals in Chinese Han nationality of East China with Goldeneye™ DNA ID 22NC Kit, allele frequencies, population genetics parameters and linkage disequilibrium information of the 21 autosomal STR were statistically analyzed. RESULTS: In the 21 autosomal STR, no deviations from Hardy-Weinberg equilibrium were detected and all loci were independent form each other. DP values of 21 autosomal STR were all above 0.85, and the combined discrimination power was 1-3.616 5 x 10(-26). Combined mean exclusion chance of this system in duo cases was 1-2.786 81 x10(-6), in trio cases was 1-8.545 82 x 10(-1). CONCLUSION Twenty-one autosomal STR included in Goldeneye™ DNA ID 22NC Kit are highly polymorphic in the Han nationality. Combined with Goldeneye™ DNA ID 20A Kit, the kit can satisfy the needs for full-sibling testing and facilitate the solution of this kind of case tools.
Alleles ; Asian People/genetics* ; China ; Ethnicity/genetics* ; Forensic Genetics/methods* ; Gene Frequency ; Genetic Loci/genetics* ; Genetic Markers/genetics* ; Genetics, Population ; Genotype ; Humans ; Polymorphism, Genetic ; Reagent Kits, Diagnostic

OBJECTIVETo investigate the expression status of 11 different cancer/testis (CT) antigen genes in esophageal carcinoma.

METHODSEsophageal carcinoma tissue and adjacent normal esophageal mucosa taken from 35 esophageal carcinoma patients were assayed for the expression of 11 different CT antigen genes by RT-PCR techniques.

RESULTSOf the 11 CT antigen genes analyzed, none of them was expressed in normal esophageal mucosa. MAGE-3 was found to be the most frequently expressed in esophageal carcinoma tissues (62.9%), followed, in the order of expression frequency, by MAGE4 (31.4%), LAGE-1 (28.6%), MAGE-1 (25.7%), CT10 (20.0%), NY-ESO-1 (20.0%), CT7 (5.7%) and SCP1 (2.9%). No expression of SSX-1, SSX-2 and SSX-4 was found. Among the 35 cases, 28 (80.0%) expressed at least one CT antigen gene, 21 (60.0%) expressed more than 2 CT antigen genes, and 4 of the 21 (19.0%) expressed more than 4 CT antigens, which accounted for 11.4% of total number of patients (4/35). No CT antigen expression was found in the tumor tissue in 7 cases, including 5 cases in stage II and 1 case each in stage I and IV, respectively. Of the 11 CT genes examined, expression of 5 genes (NY-ESO-1, LAGE-1, MAGE-1, MAGE-3 and MAGE-4) was correlated with tumor progression. SCP-1 and CT10 expression was found more frequently in early stage patients. With progression of the disease, the frequency of co-expression of multiple CT antigen genes was significantly increased reaching 28.6% in stage III patients.

CONCLUSIONOf the 11 different CT antigen genes examined by RT-PCR in esophageal carcinoma, 8 genes were detected in various frequencies in 28 of the 35 esophageal cancer patients studied. They are candidate tumor-associated antigens in the preparation of tumor vaccines for immunotherapy in esophageal cancer patients.

Adult ; Aged ; Antigens, Neoplasm ; biosynthesis ; genetics ; Base Sequence ; Esophageal Neoplasms ; genetics ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Neoplasm Proteins ; biosynthesis ; genetics