Objective To investigate the frequencies of human leukocyte antigen ( HLA)-DRB1 alleles of Chinese Marrow Donor Program (CMDP) donors in seven markets of Jiangsu Han population. Methods 2787 donors from CMDP Jiangsu Branch were typed in HLA-DRB1 locus using polymerase chain reaction-sequence-based typing (PCR-SBT) method or polymerase chain reaction-reverse sequence specific oligonucleotide probes (PCR-rSSOP) method. Frequencies of HLA-DRB1 were separately calculated in seven markets and a comparison was made among them. Results A total of 38,33,26,36,37,31,26 alleles in DRB1 locus were separately found in Huai-an, Xuzhou, Taizhou, Suqian, Nanjing, Changzhou, Yangzhou population. The most frequent HLA-DRB1 alleles in Huai-an, Xuzhou, Suqian respectively were DRB1 * 07:01, 09:01, 15:01,12:02; In Nanjing and were DRB1 * 09:01,07:01, 15:01, 12:02; In Yangzhou were DRB1 * 09:01,07:01,15:01,08:03; In Taizhou were DRB1 *09:01, 07:01, 04:05, 15:01; In Changzhou were DRB1 *09:01,07:01, 12:02, 08:03. Conclusion The above results reflected the distribution of HLA-DRB1 alleles in seven markets of Jiangsu Han population, which held abundant polymorphism, and a certain uniqueness in every market. The HLA-DRB1 allele distribution of Nanjing, Yangzhou were the closest groups to northern China, Huaian, Xuzhou, Suqian, Changzhou, Taizhou were close to northern China except DRB1 * 09:01,07:01 in different frequencies. These findings could guide the selection of future donor in CMDP Jiangsu Branch. It also could serve as foundation for HLA related disease research, studies of population genetics and anthropology in Chinese Han population.

Objective To analyze the direct and indirect effects of influencing factors on suicide ideation among college students based on the structural equation model. Methods 1505 college students were investigated with ASLEC, Simplified Coping Style Questionnaire, SSRS, QSA and BDI. Results Incidence rate of college students' suicidal ideations during the past year was 6.67%. The goodness of fit for the structural equation model was satisfactory and 3 major indices( x2/df = 2.23, GFI = 0. 982, RMSEA = 0. 029 ) had met corresponding requirements. The depression was directly influencing factor on suicide ideation, while four factors including passive coping style, social support, positive coping style and suicide attitude, had indirect impacts. Negative life event not only directly affected suicide ideation, but also had indirect effects. According to the percentages of their contribution, the risk factors were ranked as follows:depression (41.08%), negativity life event (35.35%) and passive coping style (6.05%). Similarly, the top protective factor was: social support ( 11.94% ), followed by positive coping style (4.94%) and suicide attitude (0.63%). Conclusion Depression is an important risk factor of suicide,and has a direct impact. So, not only strengthen the mental health of college students, but also train students to face up the difficulties with a positive style, and make the college students get social supports sufficiently.

In this study, a high performance liquid chromatography coupled with multi-stage mass spectrometry method was developed for simultaneous characterization of alkaloids and flavonoids, the main active components, in Kushen with diverse physical and chemical properties. Forty-two major constituents, including sixteen Kushen alkaloids and twenty-six Kushen flavonoids were tentatively identified. Additionally, useful and characteristic fragmentation pathways of two types of Kushen alkaloids, namely cytisine-type and sparteine-type, in positive ions mode were proposed and summarized, which would lay a foundation for the rapid identification of the active ingredients in traditional Chinese medicine Kushen.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods

Objective To study the changes in interleukin-32 (IL-32) in rats with Klebsiella bacillus pneumonia and approach its significance. Methods Seventy-two Sprague-Dawley(SD)rats were divide into control group,model group and experimental group by the method of random digits table,then the experimental group was subdivided into 4 hours and 1,3 and 5 days experimental subgroups(each n=6). The rat model of Klebsiella bacillus pneumonia was established by injection of 0.3 mL Klebsiella bacterial suspension into the trachea. Before the establishment of the model in the experimental group,IL-32 inhibitory agent,protease activated receptor-2(PAR2) was injected into the abdominal cavity. After model establishment,at different time points,blood was collected via tail vein to observe the changes in serum levels of IL-32,tumor necrosis factor-α(TNF-α),IL-6 and IL-8 in all the groups. The lungs were removed and stained with hematoxylin-eosin(HE)method to investigate the histopathological changes of the lung tissues under the light microscope. Results Compared to the control group, with the prolongation of time the levels of IL-32,TNF-α,IL-8 and IL-6 were increased gradually in the model group,and reached their peaks at 3 days〔IL-32(ng/L):84.40±28.24 vs. 18.57±3.86,t=5.544,P=0.002;TNF-α(ng/L):79.27±14.64 vs. 17.82±3.86, t=9.994, P=0.000;IL-8(ng/L):55.85±10.90 vs. 16.66±3.76,t=8.544, P=0.000;IL-6(ng/L):56.65±2.57 vs. 28.48±2.11,t=19.693,P=0.000〕;PAR2 could inhibit above indexes significantly,there was statistical difference at 3 days compared with the model group〔IL-32(ng/L):54.13±6.68 vs. 84.40±28.24,t=2.560,P=0.046;TNF-α(ng/L):49.12±3.56 vs. 79.27±14.64,t=4.901,P=0.003;IL-8 (ng/L):22.95±2.52 vs. 55.85±10.90,t=7.204,P=0.000;IL-6(ng/L):36.49±2.63 vs. 56.65±2.57,t=13.443, P=0.000〕. Under the light microscope,the inflammatory changes in the lung tissue in experimental group were milder than those in the model group. Conclusion As a pro-inflammatory cytokine,IL-32 can induce the production of TNF-α,IL-6 and IL-8,and the inhibition of IL-32 production may play a role in suppression of the development of Klebsiella bacillus pneumonia.

Objective It remains to be confirmed whether tissue kallikrein has neuroprotective effect in diabetes-induced stroke.This study was to investigate the neuroprotection of tissue kallikrein against cerebral ischemia-reperfusion injury in diabetic rats. Methods Healthy male SD rats were randomly divided into a sham operation, a saline control, and a tissue kallikrein group.Diabetes mellitus was induced in the animals by intraperitoneal injection of streptozotocin and the model of focal cerebral ischemia-reperfusion was made with an intraluminal vascular occlusion method. At 24 hours after modeling, we obtained the neurological deficit score, in-farct size, and brain water content, counted Iba1-and MPO-positive cells by immunohistochemistry, and determined the expressions of ICAM-1 and VCAM-1 by real-time PCR. Results In comparison with the saline controls, the rats treated with tissue kallirein showed significant decreases in the neurological deficit score (P<0.01), the infarct size ([23.57 ±5.79] vs [47.97 ±1.19]%, P<0.01), brain edema ([81.73 ±2.10] vs [84.94 ±2.34]%, P<0.05), the counts of Iba1-and MPO-positive cells (12.33 ±4.46 vs 31.83 ±8.13 and 13.83 ±4.49 vs 37.50 ±7.64, both P<0.01), and the expressions of ICAM-1 and VCAM-1 (both P<0.05). Conclusion Tissue kallikrein has a neuroprotective effect against cerebral ischemia-reperfusion injury in diabetic rats, which may be associated with its anti-inflammation property.

Objective To determine the significance of the quantitative parameters obtained from intravoxel incoherent motion ( IVIM) diffusion weighted imaging ( DWI) in differentiating renal tumors from normal renal tissues.Methods Twenty-four patients with surgical pathology-proven renal tumors and 13 volunteers with healthy kidneys were included.DWI was performed with 9 b-values (0, 20, 50, 100, 200, 400, 600, 800 and 1 000 s/mm2).The slow component of diffusion (Dslow), fast component of diffusion ( Dfast ) and fraction of fast ADC ( f) of the biexponential DWI were calculated for the clear cell renal cell carcinoma (CCRCC), the normal renal parenchyma and the non CCRCC ( NCCRCC) .The ADC was calculated for all b-values using linear regression yielding standard ADC ( ADCtot ) .The parameters were compared among the groups, and the receiver operating characteristic ( ROC ) analysis was performed. Results CCRCC showed higher ADCtot (1.73 ±0.43) ×10 -3 mm2/s, Dfast (14.75 ±14.73) ×10 -3 mm2/s, Dslow(1.34 ±0.38) ×10 -3 mm2/s than NCCRCC (ADCtot(1.23 ±0.26) ×10 -3 mm2/s, Dfast(9.47 ± 5.27) ×10 -3 mm2/s, Dslow(0.58 ±0.15) ×10 -3 mm2/s), and the differences of ADCtot(P=0.037) and Dslow(P=0.001) were significant.The normal renal parenchyma showed higher ADCtot (2.25 ±0.11) × 10 -3 mm2/s, Dslow ( 1.74 ±0.17 ) ×10 -3 mm2/s, f ( 35.00% ±9.37%) than CCRCC ( f, 31.13% ± 10.75%) and NCCRCC(f, 33.76%±24.02%), and the differences between the normal renal parenchyma and CCRCC of ADCtot ( P =0.000 ) and Dslow ( P =0.001 ) were significant.There were no differences between the normal renal parenchyma and the tumor ipsilateral renal parenchyma of all parameters.Dslow had higher accuracy ( sensitivity 95%, specificity 100%) in distinguishing CCRCC and NCCRCC, with area under the curve of 0.988.Conclusions Multi-b-value DWI derived quantitative parameters including ADCtot and Dslow may differ significantly between the renal tumor and normal renal parenchyma.Dslow is the best parameter in distinguishing CCRCC and NCCRCC.

Objective To analyze the frequencies of HLA-DQA1 alleles and their clinical values in the donor-recipient HLA-A,-B,-C,-DRB1,-DQB1 (10/10) matched hematopoietic stem cell transplantation (HSCT).Methods This study recruited 127 patients who received allogeneic HSCT and 127 unrelated donors.High-resolution (High Res) DNA typing for HLA-DQA1 alleles were performed on the 254 subjects by using sequence specific oligonucleotide probes (SSOP) and high resolution of sequence specific primer(High Res SSP).Results The DQA1 allele genotypes of 36 pairs of donor-recipient were directly identified by using SSOP.The ambiguous DQA1 allele genotypes of the rest 91 pairs were identified by using High Res SSP.Among the 127 pairs of donor-recipient,5 pairs were HLA-DQA1 alleles mismatched,while the others were all matched.No significant differences in the distribution of HLA-DQA1 alleles were observed between the donors and the recipients.Sixteen HLA-DQA1 alleles were detected in the 127 donors,which were DQA1 * 02 ∶ 01 (19.3％),DQA1* 01 ∶ 02(19.3％),DQA1 * 03 ∶ 02/03 (17.0％),DQA1 *01∶03 (9.8％),DQA1*06∶01(9.1％),DQA1*05∶ 01(7.1％),DQA1*05∶05(5.9％),DQA1*03∶01 (4.7％),DQA1*01 ∶04(2.4％),DQA1*01∶05(2.0％),DQA1*01∶01(1.2％),DQA1*05 ∶ 03(0.8％),DQA1 *05 ∶ 08(0.8％),DQA1*04 ∶ 01(0.4％),DQA1*05 ∶ 06(0.4％) from high to low frequency.Moreover,a new allele was detected in the patients.The haplotypes' frequencies and linkage disequilibrium(LD) analysis of HLA-DQA1 and HLA-DQB1 showed that the most common haplotype was DQA1 *02 ∶ 01-DQB1 *02 ∶ 02(16.1％),followed by DQA1 *03 ∶ 02/03-DQB1 *03 ∶ 03 (11.8％)and DQA1 *01 ∶ 03-DQB1 * 06 ∶ 01 (9.1％).Stronger LD were observed between DQA1 * 02 ∶ 01 and DQB1*02 ∶ 02,DQA1 *03 ∶ 02 and DQB1*03 ∶ 03,DQA1 *01 ∶ 03 and DQB1*06 ∶ 01,HLA-DQA1*06∶01 andDQB1*03 ∶ 01,DQA1*05 ∶ 01 and DQB1*02 ∶ 02(P＜0.001).Conclusion There was strong linkage disequilibrium between HLA-DQA1 and HLA-DQB1 genes.The polymorphism of HLA-DQA1 gene was less than that of HLA-DQB1 gene.No more guidance was provided to donor selection in unrelated donor-recipient HLA matched HSCT by adding HLA-DQA1 genotyping,but it might have clinical application values in HSCT with HLA Ⅱ locus mismatched donor and recipient.

With DQOL (diabetes quality of life) scale, quality of life was evaluated before and after diabetic education in 136 type 2 diabetic patients. The Cronbach′s ? of DQOL scale was from 0.809 to 0.849, suggesting that the DQOL scale did effectively reflect the life quality of type 2 diabetic patients.

Hydroxymethyltransferase (SHMT) and tryptophanase (TPase) are key enzymes in biosynthesis of L-tryptophan. We constructed three recombinant plasmids, including pET-SHMT, pET-TPase, and pET-ST for over-expression or co-expression of SHMT and TPase in Escherichia coli BL21 (DE3). The SDS-PAGE analysis showed that the recombinant proteins of 47 kDa and 50 kDa were expressed of pET-SHMT and pET-TPase, respectively. As compared to the host stain, the enzyme activity of SHMT and TPase was increased by 6.4 and 8.4 folds, respectively. Co-expression of both recombinant proteins, 47 kDa and 50 kDa, was also successful by using pET-ST and the enzyme activities were enhanced by 6.1 and 6.9 folds. We designed two pathways of dual-enzymatic synthesis of L-tryptophan by using these recombinant strains as source of SHMT and TPase. In the first pathway, the pET-SHMT carrying strain was used to catalyze synthesis of L-serine, which was further transformed into L-tryptophan by the pET-TPase expressing strain. These two steps sequentially took place in different bioreactors. In the second pathway, the pET-ST carrying strain, in which two enzymes were co-expressed, was used to catalyze simultaneously two steps in a single bioreactor. HPLC analysis indicated a high yield of 41.5 g/L of L-tryptophan was achieved in the first pathway, while a lower yield of 28.9 g/L was observed in the second pathway. In the first pathway, the calculated conversion rates for L-glycine and indole were 83.3% and 92.5%, respectively. In the second pathway, a comparable conversion rate, 82.7%, was achieved for L-glycine, while conversion of indole was much lower, only 82.9%.
Escherichia coli ; enzymology ; genetics ; metabolism ; Gene Expression Regulation, Bacterial ; physiology ; Gene Expression Regulation, Enzymologic ; physiology ; Genetic Vectors ; genetics ; Glycine Hydroxymethyltransferase ; biosynthesis ; genetics ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Recombination, Genetic ; genetics ; Tryptophan ; biosynthesis ; Tryptophanase ; biosynthesis ; genetics

Objective To introduce the operation skills of tail vein injection in conscious rats , and improve the success rate of ingection .Methods The rat was fixed by the operator with a self-made binding clothes , one person assists to fix the rat tail, one person performs puncture , and one person performs injection .During the injection process , we should minimize the injury to the rat tail and the stress caused by operation , strictly limit the needle point , the number of puncture , the maximum dose and injection speed , and to make the needle position away from the injection site to avoid contamination and waste of the drug solution .Results The operation method was successfully established and it was fast , stable, with good repeatability and high degree of coordination .Conclusion This operation is rapid, reliable and stable, worthy of recommendation , especially for the intravenous injection of expensive drugs .