Objective To evaluate the efifcacy and safety of hemostatics of injected gelatin matrix (HIGM) under the guidance of contrast-enhanced ultrasound (CEUS) for treating splenic trauma in canine model. Methods A total of 24 commercial hybrid dogs underwent celiotomy with creation of uniformly blunt splenic trauma lesion of 4.0 cm×4.0 cm×2.5 cm (length, width and depth, respectively) by hemostatic clamp. Subjects were prospectively randomized into two groups. The treatment group was treated with HIGM under the guidance of CEUS and the positive control group received thrombin solution. Conventional ultrasound and CEUS were performed to record the ascites and the splenic lesion areas at 1st, 3rd, 7th, 14th and 21st day. The ifne needle biopsy and splenectomy were performed for histopathologic examination. The weight, free intraperitoneal lfuid and injury site were compared with t test between HIGM and postive group. Results All animals in two groups survived. All dogs stopped hemorrhage after injection of HIGM under CEUS guidance. The area of injury site was (12.91±0.89) cm2, (4.45±0.75) cm2 and (1.38±0.23) cm2 at 1st, 3rd and 7th day and splenic lesions were not found at 14th and 21st day in all dogs (n=12) of HIGM group. The splenic lesion was (16.74±0.91) cm2, (11.26±0.99) cm2, (8.02±0.82) cm2 and (1.58±0.36) cm2 in the postive group at 1st, 3rd, 7th and 14th day and splenic lesions were not found at 21st day in all dogs (n=12). At 7th and 14th day post-injection, lesion areas were statistically significant between two groups (t=27.162, P=0.008;t=15.129, P=0.001). Free intraperitoneal lfuid was (0.91±0.05) cm at 1st day detected by conventional ultrasound and free intraperitoneal fluid was not found at 3rd, 7th, 14th and 21st day in all dogs (n=12) of HIGM group. The free intraperitoneal fluid in thepositive group was (1.96±0.17) cm, (1.30±0.11) cm and (0.81±0.12) cm at 1st, 3rd and 7th day and free intraperitoneal lfuid was not found at 14th and 21st day in all dogs (n=12). At 1st, 3rd and 7th day post-injection, free intraperatitoneal lfuid was statistically significant between two groups (t=20.934, P=0.003; t=41.310, P=0.000; t=22.520, P=0.000). Histopathological examination showed that there was no foreign body and foreign body granuloma and the structure of red pulp was recovered at 7th, 14th and 21st day. Gross anatomy showed that the splenic injury site was recovered completely without complications. Conclusion This study explored the value of HIGM for splenic trauma and provided a preliminary experimental evidence for clinical treatment.

To analyze killer immunoglobulin (Ig)-like receptor (KIR) gene content and allelic polymorphism in Zhejiang Han population, samples were genotyped by polymerase chain reaction sequence-specific primers (PCR-SSP). The results demonstrated that all 17 KIR genes could be observed in the population. All individuals contained 2DL4, 3DL2 and 3DL3 genes. The frequencies of these genes was 1.00. 2DL1, 2DL3, 2DP1, 3DP1*003, 3DL1, 2DS4*001/002 loci were more common, their frequencies were 0.902, 0.902, 0.902, 0.902, 0.7598, 0.5615 respectively, while the frequencies of 2DL2, 2DL5A, 2DL5B, 2DS1, 2DS2, 2DS3, 2DS4*003, 2DS5, 3DS1 and 3DP1*001/002 were relatively lower. The A KIR haplotype was the most prevalent (74.7%) in Zhejiang Han population and there were 12 different KIR haplotypes in total, the most common was 2 (53.0%). Twenty six different genotypes have been found in the population, AJ (2, 2) and AF (1, 2) showed higher frequencies, followed by AH (2, 5), NN2 (2, 6), AI (1, 5) and AG (1, 1). Fifteen of these genotypes have not been found in Caucasians so far and four new KIR profiles could not be assigned to the haplotypes according to standard assign method. In conclusion, there are distinctive frequencies of KIR gene content, haplotype as well as genotype in Zhejiang Han population.
China ; Gene Frequency ; Genotype ; Haplotypes ; Humans ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Receptors, KIR ; genetics

The purpose of this study was to establish beta block matching technique. DNA was extracted from whole blood by salting-out method, beta block matching was performed by PCR and GeneScan technique. The results showed that the length of fragments amplificated in 100 samples was different and the range of them was 91-197 bp. Amplification fragments could be divided into four regions: 91-93, 105-113, 125-139 and 177-197 bp respectively. 91 bp DNA fragments could be found in all of samples. The numbers of DNA fragments with different length have been shown high polymorphism and they focused on the range of seven to twenty four. In conclusion, the beta block matching technique is reliable and applicable to the selection of hematopoietic stem cell transplantation donors.
DNA ; genetics ; HLA-B Antigens ; genetics ; HLA-C Antigens ; genetics ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Hematopoietic Stem Cell Transplantation ; Histocompatibility Testing ; methods ; Humans ; Polymerase Chain Reaction ; methods

OBJECTIVETo study the role of contrast-enhanced ultrasound (CEUS) in the management of splenic trauma

METHODSForty patients with splenic trauma underwent CEUS examination and then different management were provided according to the examination results and the general status of patients: conservative treatment (group I, n = 19); CEUS-guided injective therapy (group II, n = 6); and surgery (group III, n = 15).

RESULTSEighteen patients were cured in group I and one patient experienced rehaemorrhagia. The haemostatic effect of CEUS-guided injective therapy was obvious in all six patients in group II. Among them, one patient experienced arteriovenous fistula, which was resolved after one week of injective therapy. Fifteen patients in Group III underwent surgery and were cured.

CONCLUSIONCEUS can provide reliable information for therapy mode selection in patients with splenic trauma and can be used to guide injective therapy.

Contrast Media ; Humans ; Spleen ; diagnostic imaging ; injuries ; Ultrasonography ; Wounds, Nonpenetrating ; diagnostic imaging ; therapy

To establish delta block HLA-matching technique, DNA was extracted from whole blood by salting-out method, delta block was amplified by polymerase chain reaction (PCR), and PCR product was detected by GeneScan. The results showed that delta block had polymorphism in 104 samples without sibship of the Han people from Zhejiang province. The range of DNA fragment length was 81-393 bp and could be divided into 4 groups: 81-118 bp, 140-175 bp, 217-301 bp, 340-393 bp. The numbers of DNA fragments were 6-32. It is concluded that the method of delta block matching is reliable and can be applied to select donors for the patients to be transplanted. It is the first time to get delta block data of the Han people in China.
HLA-A Antigens ; genetics ; immunology ; HLA-B Antigens ; genetics ; immunology ; HLA-DQ Antigens ; genetics ; immunology ; HLA-DR Antigens ; genetics ; immunology ; HLA-DRB1 Chains ; Hematopoietic Stem Cell Transplantation ; Histocompatibility Testing ; methods ; Humans

The aim of this study was aimed to investigate the molecular genetic basis for a novel HLA allele, HLA-B*4061, in Chinese population. DNA was extracted from whole blood by salting-out method. The HLA-B exons 1 - 8 of the proband was amplified and the amplified product was cloned using TOPO TA cloning sequencing kit to split the two alleles apart. Both strands of exons 2, 3 and 4 of chosen colonies were sequencing. The PCR-SSP was performed to confirm the mutations detected by sequencing. The sequencing results showed HLA-B alleles of the proband as B*4601 and the novel allele. The sequences of the novel allele have been submitted to GenBank (DQ089628, DQ089629, DQ089630). After HLA blast analysis, the novel allele showed a single nucleotide mismatch with B*400101 in exon 2 at position 272 C-->A, as the results, changing amino acid from Ser to Tyr at codon 67. It is concluded that this allele is a novel one and has been officially named B*4061 by the WHO Nomenclature Committee.
Alleles ; Amino Acid Substitution ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; HLA-B Antigens ; genetics ; immunology ; Histocompatibility Testing ; Humans ; Molecular Sequence Data ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid

BACKGROUNDThe non-operation treatment of intra-abdominal trauma guided contrast enhanced ultrasound (CEUS) is one of the hottest research topic. Gelatin/thrombin/calcium (GTC) was developed as a novel haemostatic agent for non-operable intra-abdominal trauma. We hypothesized that GTC can achieve haemostasis (without the use of pressure) within a short time in a large wound model by percutaneous injection under CEUS guidance.

METHODSForty Wister rats received large liver injuries by haemostatic clamp and were randomly divided into four groups, according to the haemostatic agent used. These included normal saline (NS) group A, lyophilising thrombin powder (LTP) group B, GTC group C, and absorbable α-cyanoacrylate (ACNA) group D. Each injury site was treated with one of the above materials and total bleeding time was recorded. All liver wounds were evaluated using CEUS at three periods: pre-injury, injury and post-treatment. The liver wounds were also evaluated by histology 3, 6, and 9 days after injury and the extents of abdominal adhesions were recorded.

RESULTSThe sensitivity of CEUS (100%) in detecting blunt traumatic liver lesions was significantly higher than conventional ultrasound (42.5%). Bleeding times at the injury site in the GTC group C ((129.3 ± 14.0) seconds) and ACNA group D ((5.2 ± 1.0) seconds) were significantly shorter than those in the NS group A ((369.5 ± 48.8) seconds, P < 0.01) and LTP group B ((324.7 ± 52.22) seconds, P < 0.01). The LTP group B showed no significant difference compared with the NS group A. Gross examination of liver tissue revealed that there were fewer intra-abdominal adhesions in the GTC group C (10%) than in the ACNA group D (100%). Histopathologic examination showed that GTC was completely absorbed after nine days.

CONCLUSIONSGTC, delivered by percutaneous injection under CEUS, may achieve haemostasis (without the use of pressure) within a short time in a large wound model. GTC is absorbable and may prevent intra-abdominal adhesions. Therefore, it may be the optimal choice for first aid treatment of large abdominal wounds in the setting of blunt trauma.

Animals ; Calcium ; administration & dosage ; therapeutic use ; Gelatin ; administration & dosage ; therapeutic use ; Hemorrhage ; diagnostic imaging ; drug therapy ; Hemostatics ; administration & dosage ; therapeutic use ; Injections ; Liver ; diagnostic imaging ; injuries ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Thrombin ; administration & dosage ; therapeutic use ; Ultrasonography

OBJECTIVETo investigate the mRNA expression profiles of megakaryocytes (MKs) from human cord blood CD34+ cells ex vivo expanded using Solexa technique.

METHODSCD34+ Cells were isolated using density gradient centrifugation and magnetic activated cell sorting. Cultures were stimulated with recombinant human thrombopoietin (100 ng/ml). After 12 days, the MKs fraction was separated from the non-MKs fraction using an anti-CD41 monoclonal antibody by immunomagnetic sorting. The mRNA expression of MKs and non-MKs was detected by Solexa sequencing.

RESULTSWe obtained 3 773 147 and 3 533 805 Tags from MKs and non-MKs, respectively. The amounts of unambiguous tags were 3 291 132 and 2 967 947 and those of distinct tags were 197 769 and 245 318. The expression of 1161 genes was up-regulated and that of 902 genes down-regulated. The expression of 2717 tags was up-regulated and that of 1519 tags down-regulated.

CONCLUSIONSMKs and non-MKs have remarkably different mRNA expression profiles. The differential gene-encoded products may be involved in cellular development, adhesion, apoptosis metabolism, intra- and intercellular signal transduction, and immune response. Further studies on this topic may clarify the expression mechanism, signal transduction, and regulation mechanisms.

Antigens, CD34 ; Cells, Cultured ; Fetal Blood ; cytology ; Humans ; Megakaryocytes ; cytology ; metabolism ; RNA, Messenger ; genetics ; Transcriptome

ObjectiveTo explore the effects of ischemic preconditioning on the level of serum testosterone (T) and apoptosis of spermatogenic cells in rabbits with testicular ischemia-reperfusion injury induced by testicular torsion.

METHODSA total of 15 New Zealand male rabbits were randomly divided into groups A (control), B (ischemia-reperfusion), and C (ischemic preconditioning). The animals of group A were subjected to exposure of the right spermatic cord without ischemia, those of group B received 60-minute non-invasive occlusion of the right spermatic cord followed by 3 days of reperfusion, and those of group C underwent 5-minute occlusion plus 5-minute reperfusion of the right spermatic cord followed by the same procedure as that for group B. Then the rabbits were narcotized with 3% barbital sodium, the whole blood collected for examination of the serum T content and the testis tissues obtained from both the ischemic and healthy sides for HE and TUNEL staining.

RESULTSAfter operation, the body weight was significantly increased as compared with the baseline in groups A (［2.65±0.07］ vs ［2.45±0.07］ kg, P<0.05) and C (［3.03±0.11］ vs ［2.92±0.07］ kg, P<0.05), but not in group B (［3.05±0.07］ vs ［3.05±0.07］ kg, P>0.05). The serum T level showed no statistically significant difference in group A before and after operation (［139.59±9.39］ vs ［140.19±9.47］ ng/L, P>0.05), but was remarkably lower after operation than the baseline in groups B ［148.06±3.31］ vs ［74.12±4.00］ ng/L, P<0.01) and C (［133.75±6.48］ vs［94.76±3.13］ ng/L, P<0.01) as well as than the postoperative index in group A (P<0.01). In comparison with group A and the healthy side of group B, the testis tissue of the ischemic side in group B exhibited structural damage of most of the seminiferous tubules with disappearance of spermatogenic cell structures, apoptosis of spermatogenic cells, and exudation of light-eosin edema fluid in the mesenchyme and lumen, with a markedly increased apoptosis index (P<0.01) and a significantly decreased Johsen's score (P<0.01). Compared with ischemic side of group B, The testis tissue of the ischemic side in group C was restored to normal as compared with that in group B, with a dramatically decreased apoptosis index (P<0.01) and a remarkably increased Johnsen's score (P<0.01).

CONCLUSIONSIschemic preconditioning can raise the decreased serum T level and reduce the apoptosis of spermatogenic cells in rabbits with testicular ischemia-reperfusion injury, which could be applied as a potential option for the clinical treatment of testicular ischemia-reperfusion injury.

Animals ; Apoptosis ; Germ Cells ; In Situ Nick-End Labeling ; Ischemia ; physiopathology ; Ischemic Preconditioning ; Male ; Rabbits ; Random Allocation ; Reperfusion Injury ; blood ; physiopathology ; Spermatic Cord Torsion ; Testis ; physiopathology ; Testosterone ; blood

The study was purposed to investigate the molecular genetic basis for HLA novel allele HLA-B*5408N in Chinese population. DNA was extracted from whole blood by commercial DNA extraction kit, the HLA-B exons 2 - 4 of the proband was amplified by allele specific primers PCR and the amplified product was sequenced for exons 2, 3 and 4 bidirectionally. The sequencing results showed HLA-B alleles of the proband as B*1527 and the novel allele. The sequences of the novel allele have been submitted to Genbank (DQ295998, DQ295999, DQ296000). After blast analysis, the novel allele showed a single nucleotide mismatch with HLA-B*5401 in exon 3 at position 553 G-->T, which resulted in an amino acid changing from Glu to premature stop codon at position 161. No the HLA-B54 antigen specificity expression in the proband cells was found using HLA-AB serological Typing Trays. It is concluded that this allele is a novel null allele and has been officially named B*5408N by the WHO Nomenclature Committee.
Alleles ; China ; Exons ; genetics ; HLA-B Antigens ; genetics ; Humans ; Polymerase Chain Reaction ; Sequence Analysis, DNA