Objectives:The purpose of this study is to provide a HLA B locus genotyping method which is expected to compensate the unsatisfactory serology B locus typing and to explore the distribution of B22 subtypes.Methods:Taking standard cell lines provided by the XIIth International HLA Workshop as reference,the authors established the PCR SSP method for HLA B genotyping.By this method,the HLA B alleles of leukemia patients were typed and 57 individuals previously identified as HLA B22 by serologic typing were genotyped.Results:The results of B locus genotyping of 104 cell lines by PCR SSP and that of reported were completely concordant .Unambiguous results of 17 leukemia patients and their relations were gotten except one discordant from serology which was thought as mistaken serotyping .Out of 57 samples ,55 were confirmed to bear B54 or B55 or B56 allele,respectively ,with B54 being the most common allelic form,and another showed a unique amplifying pattern which was different from any known HLA B22 alleles so far reported, suggesting a new allele or "mismatched ”DNA double strands which needed further study .Conclusions:PCR SSP was proven to be a practical genotyping method for B locus because of its simplicity, rapidity ,accuracy and unvariability with changes of health.

Specimens of skin, skeletal muscle, adipose tissue, brain, peripheral nerve and catilage were taken from 6 cadavers (divided into 2 groups) perfused and embalmed with formalin. 90 pieces of the above-mentioned tissues were taken in different times and the paraffin sections were stained by routine HE. Electron microscopic observation was made from some skin and skeletal muscle specimens, and histochemical methods were used to demonstrate DNA, RNA, keratin(-SH, -SS), fatty acids and calcium in some skin materials. Loyez stain method for myelin sheath was applied to some brain and nerve sections.1. Skin tissue: In both group A and B, 83.3% and 78.9% of skin materials were appraised as grade I and II (excellent and good) respectively. The morphological changes appeared mainly as karyopyknosis of the epidermal cells and shrinkage of the secretory part of the sweat glands. Most of the skin tissues of the corpses perfused 3～4 hrs after death were well preserved, only 8.3% being in grade Ⅲ (moderate). On the other hand, 43% of skin materials taken from those perfused 30 hrs after death were in grade Ⅲ. Ultrastructurally, the shape of the epidermal cells and their adhensions still remained intact, but karyopyknosis or chromolysis and autolysis of most of the cytoplasmic membraneous structures were found. In those cadavers preserved for less than one year, there was no evidence of progressive changes in microscopic and submicroscopic structures of the skin. The tonofillaments and desmosomes were still kept well in those specimens preserved for many years, and their presence may be the cause of maintaining cell integrity. Histochemically, mercapto-reaction of keratin was kept stable and found to be well shown even in old samples of skin, but the disulfide reaction usually showed marked fading with time, probably due to exofolition of the stratum corneum. However, this reaction was well shown in the hairs and not weakened for many years.2. Fat tissue: Hydrolysis of lipid substances of subcutancous fat and sebaceous gland, as shown by the presence of fatty acid, was found in the corpses preserved even as short as half a year.3. Skeletal muscle: The preservation state of the skeletal muscle was also dependent on the complete perfusion in time. The structure of the muscle fibers was generally worse preserved than that of the skin as shown in severe myolysis. Since stagnation of the small blood vessels frequently accompanied myolysis, obstruction of microcirculation which prevented the permeation of fomalin into the muscle tissues may be the cause of rapid myolysis. The ultrastructural changes of the muscle fibers were the same as those of the epidermal cells. In all specimens even as old as 30 years, two types of myofillaments were well preserved, and bands could be seen distinctly.4. Nervous tissue: The brain tissues were well preserved in those corpses that were perfused shortly after death, but the peripheral nerves usually showed severe autolysis, obviously it is due to the same reasons as those for myolysis.5. Cartilage: The structural changes of cartilage after death were slow and all specimens of cartilage were well preserved.

The arrangement of fascicles of musculocutaneous nerve at different levels in brachial plexus were studied in 22 adult cadavers. The nerve fascicles were dissociated and traced under operative microscope. At the distal end of the musculocutaneous nerve, the muscular and cutaneous fascicles are separated.The muscular fascicles are situated at the latero-anterior quadrant of the cross-section of the nerve.The mixed fascicle are become predominated proximally. The muscular and the muscular predominated mixed fascicles are still situated at latero-anterior quadrant. At the level 1 cm below the clevicle, 1cm and 1.5 cm above the clevicle,the fascicles of the nerve are situated at the lateral superio-anterior quadrant of the crosssection of relative parts of brachial plexus. The fascicles of the nerve are situated at the anterior quadrant of the crosssection of superior trunks. The fibers of musculocutaneous nerve are originate from C5, C6 and C7, being 31.6,％, 64.6％ and 3.8％,respectively.

Objective To study the diagnosis and treatment of infected pseudoaneurysms(PA) resulting from drug injection. MethodsThe clinical data of 11 cases of i nfected PA resulting from drug injection were reviewed retrospectively.1 pat ient was treated by non-operation,10 cases were treated by artificial blood ve ssel transplantation after removal of PA and local debridement. Results One case dead. All the 10 cases treated by operation had satisfactory results and none had ischemic symptom of the limbs.Eight grafts were patent d emonstrated by color Dopler ultrasonography 3 months postoperatively. Conclusions The diagnosis of PA induced by drug injection can be m ade by the history of drug injection, pulsate mass of injection site. The eff ective treatment should include control of infection,local debridement,PA res ection and blood vessel implantation .

Objective:To investigate the effects of nitric oxide(NO) and inducible nitric oxide synthase (iNOS) in the experimental abdominal aortic aneurysm (AAA) rat model.Methods:An intra-aortic elastase infusion model was used.Control rats received intra-aortic saline infusion.In the remaining groups,intra-aortic elastase infusion was used to induce aneurysm formation.These rats were treated with intraperitoneal injections of saline postoperatively(experimental group),aminoguanidine postoperatively(medicine group).Serum NO and aortic diameter were measured,Changes of histology,iNOS and MMP-9 were observed in the aortic wall.Results:Experimental group produced AAAs with significant production of iNOS,MMPs and serum NO compared with controls.In medicine group reduced aneurysm size and displayed suppression of MMPs expression,inflammatory infiltrates and serum NO production were detected.Conclusion:Expression of iNOS and MMP-9 are induced and serum NO levels are increased in experimental AAA,iNOS and NO production by iNOS play an important role with detrimental effects during experimental aneurysm development.

Objective To study the early functional change of sinusoid endothelial cell after liver transplantation in rat, and to investigate the endothelia protective effect of prostaglandin E_1(PGE_1). Methods Rat orthotopic liver transplantation model was performed in "two-cuff method", grouped as follows: group A served as normal rat blank control, group B as operative control with normal donor, group C as experimental control with shock donor, and group D as experimental group with shock donor and PGE_1 administration ( n =8 in each group). Transplanted groups (referring to recipients without specific definition) were sacrificed 6 h after operation for blood taken to detect serum liver enzymes (ALT, LDH), malondialdehyde (MDA), nitric oxide (NO) and plasm endothelin (ET). Liver tissue was resected at the same time for standard pathologic examination. Comparison of the difference the results was made between groups. Results Cold preservation time and anhepatic phase were similar in each group, (2?0.5) h and (15?3) min respectively. All survived 6 h after transplantation (8/8) in group B and D with a survival rate of 100%, only 5 survived 6 h after transplantation in group C (5/8) with a survival rate of 62.5%. Comparing with group C, blood ALT, LDH, MDA, ET decreased and NO increased significantly in group D ( P

In this paper, the aging of the stem cells has been elucidated in the aspect of quantitative and qualitative exhaustion, such as balance self-renewal and differentiation of hematopoietic stem cells. The reduced telomere and oxidative stress both accelerate the aging course, besides, histone code and transcriptional activation lead to the DNA or protein damage. It plays an important role in the stem cell aging that the modulation of allochromosome structure and epigenetic regulation of gene transcription. If the aging of the stem cells doesn’t happen on the level of the gene, it must result from the transcription. During the haemopoiesis process of stem cells, all kinds of the transcriptional mistakes will lead to the aging of cell cycle, which is the result of allochromosome enlarging. However, the above mechanism will not occur normally. Only allochromosome allotopia is not explainable to the gene expression of stem cells. The proof of stem cells’ self-renewal evidences that if not the consequence of cell expression, allochromosome change the histone.

Objective:To evaluate the efficacy of posterior subtotal vertebrectomy in the treatment of thoracolumbar vertebral refractures after vertebroplasty.Methods:A retrospective analysis was conducted in the 28 patients with refracture after percutaneous vertebroplasty(PVP) or percutaneous kyphoplasty(PKP) who had been treated at Department of Spinal Surgery, Henan Provincial People's Hospital from June 2017 to October 2019. They were 7 males and 21 females, with an average age of 62.4 years(from 61 to 76 years). A total of 28 vertebrae were involved, including 5 T11s, 9 T12s, 11 L1s and 3 L2s. Their previous operations were PKP in 17 cases and PVP in 11. After the spinous process, vertebral plate, articular process and transverse process were resected by posterior approach, the vertebral body, bone cement and upper and lower intervertebral discs were partially resected by trans-vertebral lateral approach. At the same time, nerve decompression was performed. Finally, the inter-vertebral support was fixated followed by the posterior screw-rod orthopedic fixation. The operation time and intraoperative bleeding volume were recorded. The cobb angles of kyphosis were compared on the X-ray films of the whole spine between preoperation and the last follow-up to evaluate correction. Functional improvement of the spine was evaluated by comparison of the visual analogue scale (VAS) and JOA(Japanese Orthopedics Association) scores between preoperation and the last follow-up.Results:The operation time averaged 182.1 min and intraoperative bleeding volume 996.2 mL. All the 28 patients were followed up for 8 to 29 months (mean, 19.8 months). No obvious neurological lesions or other serious complications were observed. The cobb angle was improved from preoperative 41.3°±10.3° to 6.4°±2.5° at the last follow-up, the VAS score from preoperative 7.3±1.8 to 2.5±1.0 at the last follow-up, and the JOA score from preoperative 8.4±2.3 to 21.3±2.5 at the last follow-up, showing a significant difference in all the comparisons ( P<0.05). Conclusion:The posterior subtotal vertebrectomy is effective for thoracolumbar vertebral refractures after vertebroplasty because it can remove bone cement, decompress the spinal canal, fuse the inter-vertebral graft and reconstruct the spinal stability in one stage.

In order to explore the pathogenic characters,diagnosis and treatment of canine leptospirosis,a retrospective analysis of the pathogenetic factor,diagnosis and treatment of three dogs were reported which diagnosed as canine leptospirosis in China Agricultural University Teaching Animal Hospital from August to November,2015.The infected dogs showed symptoms of depressed,jaundice,anorexia and vomiting.Two of them had fever history and contacted the water resource outside before getting ill.All three dogs were confirmed anemia,liver and kidney damage by laboratory examination.They were diagnosed with canine leptospirosis according to the positive results of the Leptospira PCR examination base on urine.During the diagnosis and treatment process,one of them was dead,the rest two recovered after antibiotic and symptomatic therapy.In conclusion,dogs which infected with leptospirosis may have a contact with the infected water before.The Leptospira PCR examination base on urine can be used for the diagnosis method of the disease.As to treatment,the use of penicillin and tetracycline drugs such as doxycycline is recommended.

Objective:To explore a new method of the cultivation of adoptive immunotherapy cells.Methods: Mononuclear cells was isoplated by density gradient centrifugation and then proliferated by using CD 40-agonist monoclonal antibody 5C11、cytokine of IFN-αand IL-7(CD40 group)in vitro.During the culturing procedure ,the cell morphology was obersved by optical microscope.The percentage of T-lymphocytes, NK-T cells, Treg cells and the cell proliferation, which were compared with CIK group CD3mAb activated,was detected on the 9th day.Results:There was no significant difference of CD 4+/CD8+T cells percentage between the two groups.But the Treg cells percentage of CIK group was far higher than that of CD 40 group,while the percentage of CD3+CD56+NK-T cells was lower than that of CD 40 group.And a group of Mo-NK-DC cells were observed in the CD 40 group.Conclusion: The new method of adoptive immunity therapy has been established in this study could increase the percentage of NK -T cells which had the ability to kill tumor cells.Simultaneously ,it is reduced the amount of Treg cells significantly.