Ahigh-resolutionmethodforhumanleukocyteantigen-B(HLA-B)genotypingwasestablished based on the optimized polymerase chain reaction, cloning and sequencing technology. The exon2 and exon3 of HLA-B gene were amplified with primers based on the HLA-B gene sequence. These produced heterozygous alleles were effectively cloned into plasmid DNA based on the principle of plasmid incompatibility, and were followed by bacterial culture. Then Sanger sequencing was carried out and after analyzing the result by software ClustalX2 and IMTG/HLA database comparison, the HLA-B genotype of the samples was achieved. Seven clinical samples were detected, and the results were consistent with those of PCR-SBT genotyping method. The method was cost-effective, high-resolution and it did not require technical software. The use of universal primers simplified the cumbersome design and optimization process of specific primers.

Objective To investigate whether catalpol affects senile plaque formation and spatial learning and memory ability in the amyloid-β protein precursor/presenilin 1 (APP/PSI) double transgenic mice.Methods Three month-old APP/PS1 double transgenic mice were randomly divided into catalpoltreated and saline-treated groups (n =10),with C57 mice of the same age and genetic background as normal control group (n =10).The catalpol (in a dose of 5 mg · kg-1 · d-1) and the same amount of saline were peritoneally injected into Alzheimer' s disease (AD) model mice for 3 weeks.Immunohistochemical staining was performed to examine senile plaques in the brain of AD model mice,and Morris water maze was used to assess the spatial learning and memory abilities of the mice.Results Compared with the saline-treated AD model mice (6.0 ±0.6),the number of senile plaques of catalpol treated AD mice significantly decreased (2.3± 0.7; t =3.500,P =0.025); Mice in each groups had similar latency and path length to reach platform in visible platform test; In hidden platform test,catalpol-treated mice had a significant lesser latency and path length compared with saline-treated mice,furthermore,catalpol-treated mice had much more platform-crossing times (6.4 ± 0.8) than saline-treated mice (2.9 ± 0.4 ; t =5.592,P =0.001).Conclusion Catalpol can significantly decrease the senile plaque formation and improve the spatial learning and memory abilities of APP/PS1 double transgenic mice.

Objective To compare the effects of pulse high-volume hemofiltration (PHVHF) and continuous veno-venous hemofiltration (CVVH) on severe acute panceatitis (SAP). Method From January 2005 to December 2009, a total of 38 patients with SAP were randomly(random number) divided into PHVHF group ( n = 18)and CVVH group ( n = 20). After hemofiltration for 72 hours, clinical symptoms, APACHE Ⅱ score, biochemical changes and mortality were observed. The levels of TNF-α, IL-6, and IL-10 in plasma were assayed by using ELISA before and after treatment. The doses of dopamine used in shock patients were also observed. Measurement data were expressed in(-x) ± s, and t-test was used for comparison between two groups. Results In both groups ,symptoms were markedly improved after treatment. The APACHE Ⅱ score, serum amylase, creatinine, and white blood cell count were decreased ( P ＜ 0.05). Besides, hypoxemia and acidosis were corrected, and the PHVHF group was superior to the CVVH group especially in heart rate, breathing and APACHE Ⅱ score ( P ＜ 0. 05).The levels of TNF-α, IL-6 and IL-10 decreased in both groups ( P ＜ 0.05), and the PHVHF group was superior to the CVVH group ( P ＜ 0. 01 ). The doses of dopamine used in shock patients also decreased in both groups ( P ＜0. 01 ), and they decreased more in PHVHF group than in CVVH group ( P ＜ 0.05). The mortality was 11.1%in PHVHF group and 25 % in CVVH group. Conclusions PHVHF is obviously superior to CVVH in the treatment of SAP, and can serve as an important adjuvant therapy for SAP, stabilizing the hemodynamics and reducing the levels of pro-inflammatory factors and mortality.

Objective:To analyze the clinical efficacy of Seretide combined with noninvasive positive pressure ventilation (NIPPV) in the treatment of chronic obstructive pulmonary disease (COPD) complicated with type 2 respiratory failure (RF).Methods:120 patients of COPD complicated with type 2 RF were randomly divided into the observation group and the control group.All patients were treated with routine,symptomatic and supportive treatment,based on that NIPPV was used in the control group,and the observation group was treated with Seretide on the basis of NIPPV.The levels of arterial blood gas,inflammatory factors,pulmonary function and CAPS score before and after treatment were compared and analyzed.Results:After treatment,the pH,PaO2 and SaO2 of both groups were significantly increased,PaCO2 levels weresignificantly decreased,serum IL-33,TNF-oα and sICAM-1 levels were significantly decreased,FVC,PEFR,FEV1％ and FEV1/FVC were significantly decreased,and the CAPS scores were significantly decreased;the changes of all the index mentioned above in the observation group were more statistically significant (P＜0.05).Conclusion:Seretide com bined with NIPPV could more effectively decrease the levels of inflammatory factors and improve the blood gas and lung function in the patients of COPD with type 2 respiratory failure.

Pyrosequencing is one of the important genetic polymorphism detection methods currently, but the complicated pretreatment procedure limits its application in clinical test. To simplify the whole process of pyrosequencing, on the basis of the linear_after_the_exponential_polymerase chain reaction ( LATE_PCR) , we improved the primer design method of LATE_PCR, increased the length and the concentration of the excess primer, applied direct amplification technology with whole blood, and established a whole blood_imLATE_PCR method based on common rTaq polymerase and “HpH Buffer” ( High pH buffer ) . The amplification system was optimized, and the influences of blood anticoagulant and the amount of whole blood template were investigated. The single stranded template for the pyrosequencing was obtained by PCR amplification using a single tube in one_step process, and the alcohol dehydrogenase gene polymorphisms of 24 clinical blood samples were then detected successfully. The results could be used to guide clinical individualized medication. The genotypes of ADH1B locus of 24 samples were 6 cases of AA homozygote, 14 cases of AG heterozygote, and 4 cases of GG homozygote. The genotypes of ADH1C were 20 cases of GG homozygote, 4 cases of AG heterozygote, and no cases of AA homozygote.

Objective To discuss the coverage of finger soft tissue defect with dorsal proximal digit fasciocutaneous flap on the middle and distal digit.Methods From May,2013 to December,2014,8 cases with soft tissue defects at 8 fingers were treated with dorsal proximal digit fasciocutaneous flap.The flap sizes ranged from 2.5 cm × 2.0 cm to 3.5 cm × 3.0 cm.The donor site were closed straightly.Results Eight flaps of 8 fingers survived.All the wounds at the donor sites healed well.Eight fingers in 8 cases were followed up for 6-12 months.The color,texture and contour of the flaps were satisfied.The two-point discrimination distances were 8-10 mm.Conclusion The skin defect in the middle and distal digit can be satisfied covered with dorsal proximal digit fasciocutaneous flap.This flap is a simple,reliable and safe management for digit defect and can be performed in the primary hospital.To ensure the surviving of the flap,ensure the surviving of the flap,the awareness of the anatomy of the flap should be known well.The limits of its reconstruction of sensation and coverage size exit in its application.

Objective To investigate the derivation and distribution of nitric oxide synthase (NOS) and its relation with the microvessels in the spinal cord. Methods The microscopic features and ultrastructure of NOS in the spinal cord tissue of rats were observed by immunohistochemistry and immunoelectronmicroscopy. Results NOS immunoreactive interneurons were identified mainly in the intermediolateral cell column, around the central canal and in the dorsal and ventral horn, and many of the NOS-containing neural processes and neurons were closely associated with the spinal cord microvessels. Conclusion The regional distribution of NOS may be indicative of the involvement of nitric oxide (NO) in different function related to movement, sensory processing and visceral regulation, and the ultrastructural features benefit the NO diffusion into the microvessels to regulate the blood flow in the spinal cord.

Objective To investigate the derivation and distribution of nitric oxide synthase (NOS) and its relation with the microvessels in the spinal cord. Methods The microscopic features and ultrastructure of NOS in the spinal cord tissue of rats were observed by immunohistochemistry and immunoelectronmicroscopy. Results NOS immunoreactive interneurons were identified mainly in the intermediolateral cell column, around the central canal and in the dorsal and ventral horn, and many of the NOS-containing neural processes and neurons were closely associated with the spinal cord microvessels. Conclusion The regional distribution of NOS may be indicative of the involvement of nitric oxide (NO) in different function related to movement, sensory processing and visceral regulation, and the ultrastructural features benefit the NO diffusion into the microvessels to regulate the blood flow in the spinal cord.

Objective To observe the biological activities of human doppel(Dpl) protein transiently expressed and Dpl-like protein PrP?32-121 on a human neuroblastoma cell line SH-SY5Y.Methods Recombinant mammalian expression plasmids containing human PRND gene and truncated PrP?32-121 fragment were generated by PCR.The expression and location of Dpl and PrP?32-121 post-transfection were observed by IFA.The cytotoxicity was measured by MTT analysis.Cellular apoptosis was investigated by flow cytometry and Western blot.Results Both Dpl and PrP?32-121 protein were expressed and mainly located on the cell membrane.Remarkable cytotoxicity was detected on SH-SY5Y cells after 24 h transfection.Meanwhile,more Annexin V/PI positively-stained cells as well as lower levels of cellular pro-caspase-3 and Bel-2 were detected in the cells receiving Dpl and PrP?32-121 expressing plasmids.Conclusion Dpl protein transiently expressed and PrP?32-121 can lead to the similar neural cytotoxicity,probably triggering the cell apoptosis program.

Objective To observe the biological activities of human doppel (Dpl) protein transiently expressed and Dpl-like protein PrPΔ32-121 on a human neuroblastoma cell line SH-SY5Y. Methods Recombinant mammalian expression plasmids containing human PRND gene and truncated PrPΔ32-121 fragment were generated by PCR. The expression and location of Dpl and PrPΔ32-121 post-transfection were observed by IFA. The cytotoxicity was measured by MTT analysis. Cellular apoptosis was investigated by flow cytometry and Western blot. Results Both Dpl and PrPΔ32-121 protein were expressed and mainly located on the cell membrane. Remarkable cytotoxicity was detected on SH-SY5Y cells after 24 h transfection. Meanwhile, more Annexin V/PI positively-stained cells as well as lower levels of cellular pro-caspase-3 and Bel-2 were detected in the cells receiving Dpl and PrPΔ32-121 expressing plasmids. Conclusion Dpl protein transiently expressed and PrPΔ32-121 can lead to the similar neural cytotoxicity, probably triggering the cell apoptosis program.