Objective To analyze the molecular genetic basis of novel allele HLA-B * 9534 and establish the allele group specific primer PCR method. Methods Genomic DNA was extracted from whole blood by commercial DNA extraction kit. The HLA-B exons 1 to 8 coding sequences of the proband were am-plified by PCR and the amplification product was purified with double enzymes digestion and both strands of exons 2, 3 and 4 were sequenced. The exon 2-4 amplification of the HLA-B * 9534 was performed with al-lele group specific primers PCR and the PCR product was directly sequenced for exon 2 to 4. Results The proband has two HLA-B alleles. The result was assigned for HLA-B * 1518 and B * 4601 combination with a mismatch in 593A/G heterozygote by DNA sequencing of exon 2 to 4 with loci primers. After separating the two alleles of the proband with allele group specific primers polymerase chain reaction method, HLA-B * 4601 and HLA-B * 9534 alleles were identified after sequencing. The HLA-B * 9534 is identical to HLA-B * 1518 except for one nucleotide substitutions in exon 3 at position 593 A→G, this results in amino acid substitution at cedon 174 from Asn to Ser. The sequences of the novel allele have been submitted to GenBank (EU046491) and the allele has been officially nominated by the WHO Nomenclature Committee. Conclusion Identification of a novel HLA-B * 9534 allele and allele group specific primer PCR for HLA-B * 9534 was re-liable.

Objective To investigate the effect of B7 Homology 3(B7H3)on brain damage of S .Pneumococcal(SP)meningitis . Methods SP meningitis was established by intracerebral ventricular injection of SP suspension on wild-type BALB/C mice .48 mice were divided into 4 groups and received following injections :NS(CON group) ,recombinant murine B7H3alone(B7H3 group) ,SP group ,SP+B7H3 group .At 18 ,48 ,72 h post infection ,mice were conducted neurobehavior score ,then they were anesthetized and killed by cervical vertebra dislocation ,brains were collected .The mRNA expressions of NSE and S100b were detected by real-time PCR .Results Compared with CON group ,the scores of recombinant murine B7H3 group had no significant change at 18 ,48 ,72 h after infection of SP(P>0 .05);at different time points the scores of SP group were decreased significantly than the CON group (P<0 .05);scores of SP+B7H3 group decreased furtherly than SP group(P<0 .05) .The relative expressions of NSE ,S100b mR-NA in brain tissue homogenate :for the NSE ,S100b mRNA relative expressions ,there was no significant difference between B7H3 and CON group at 18 ,48 ,72 h post SP injection(P>0 .05) .At 18h ,48h ,72h ,post infection mRNA expressions of NSE ,S100b in SP group increased compared with CON group(P<0 .05);the mRNA expressions of NSE ,S100b increased furtherly in SP+B7H3 group compared with SP group(P<0 .05) .Conclusion B7H3 upregulates the mRNA expressions of NSE and S100b ,and promotes the progress of SP meningitis in mice .

ObjectiveTo investigate the clinical features and prognosis of children with rotavirus gastroenteritis and convulsion.MethodsClinical data of children with rotavirus gastroenteritis hospitalized from January 2010 to December 2013 were retrospectively analyzed. Subjects were divided into the seizure group and no seizure group according to the presence of seizure in the course and compared between the two groups.ResultsThere were no signiifcant differences in sex, age, and the average duration of hospitalization between two groups (allP>0.05). The family history, history of seizures, the levels of serum sodium, calcium, lactate, standard bicarbonate concentration (SB), actual bicarbonate concentration (AB), carbon dioxide content (TCO2) and pH were statistically signiifcant between two groups (allP>0.05). During the follow-up period (outpatient telephone follow-up), the recurrence of seizure in two groups was signiifcant different (P<0.05) and only one (0.54%) child in seizure group developed epilepsy.ConclusionThis study showed that rotavirus gastroenteritis with convulsion is a benign clinical course.

Objective To investigate the effect of Roux-en-Y hilar-jejunum anastomosis in surgical resection of hilar cholangiocarcinoma.Methods The clinical data of 31 patients with hilar cholangiocarcinoma admitted from Feb 2009 to Feb 2017 who underwent Roux-en-Y hilar-jejunum anastomosis after resection were retrospectively analyzed.Results According to Bismuth-Corlette classification there were 6 cases of Ⅲ a,6 cases of Ⅲ b and 19 cases of type Ⅳ.These patients all successfully underwent Roux-en-Y hilar-jejunum anastomosis,with intraoperative blood loss of 50-4 000 ml [on average of (1 146 ± 1 082) ml].The average operation time was (346 ± 118) min,and the average hilar-jejunum anastomosis time was (35 ± 13) min.The average postoperative hospital stay was (20 ± 11)days.There were 25 cases of R0 resection (80.6％).The postoperative complication rate was 25.8％(8/31).The overall median survival time was 21 months,and the 1-,3-,and 5-year survival rates were 68.8％,14.6％ and 3.6％,respectively.During the follow-up period,7 cases of reflux cholangitis,5 cases of biliary anastomotic stenosis,and 1 case of adhesive intestinal obstruction were found.Conclusion The hilar-jejunum anastomosis is simple,safe,widely applicable prcedure,which is easy to perform.

Objective To investigate the clinical efficacy and prognostic factors for combined caudate lobectomy radical resection plus broad lymph node dissection in patients of hilar cholangiocarcinoma.Methods The clinical data and follow-up results of patients with hilar cholangiocarcinoma surgically treated from Feb 2008 to Feb 2017 were retrospectively analyzed.Result The R0 resection rate[72.2％ (13/18) vs 43.9％ (18/41)],operation time [(433 ± 136) min vs (302 ± 122) min],intraoperative blood loss [(1 789 ± 1 091) ml vs (776 ± 755) ml] and postoperative complication rate [66.7％ (12/18) vs 36.6％ (15/41)]were significantly higher in the hepatic lobe combined with caudate lobe resection group than that without caudate lobe resection group (P ＜ 0.05).The median survival time of patients with enlarged lymph node dissection was longer than that of patients with regional lymph node dissection (33 months vs 13 months,P ＜0.05).Univariate and multivariate analysis showed that the preoperative serum CA199 level ＞ 1 000 U/ml,the degree of microscopic margin and tumor TNM stage were significantly correlated with the prognosis of the patients (P ＜ 0.05).Conclusion Combined with caudate lobe resection can improve R0 resection rate.Targeted lymph node dissection helps prolong survival.The degree of microscopic margin,preoperative CA199 and TNM staging are independent risk factors for the prognosis of patients with hilar cholangiocarcinoma.

Objective To investigate the diagnosis and treatment of upper gastrointestinal hemorrhage caused by pancreatic segmental portal hypertension.Methods The clinical diagnosis and follow-up data of 13 patients with upper gastrointestinal hemorrhage caused by pancreatic segmental portal hypertension from Jan 2010 to Dec 2017 were retrospectively analyzed.Results Of the 13 patients,5 had pancreatic pseudocysts and 8 had chronic pancreatitis.All of them had a history of hematemesis or (and) tarry feces,and 2 of them had a history of hemorrhagic shock.13 patients had isolated gastric varices,and 5 of them had varicose veins in the lower esophagus.13 patients had splenomegaly and hypersplenism,and all patients underwent splenectomy.All patients were followed up,and the varicose veins were significantly improved or disappeared.During 1 year to 8 years of follow-up,there was no rebleeding.Conclusion Splenectomy cures upper gastrointestinal hemorrhage caused by rupture of the varicose veins in patients of pancreatic segmental portal hypertension.Preoperative interventional spleen artery embolization can reduce the difficulty of spleen resection.

Objective To evaluate the therapeutic effect or safety of selective paraesophagogastric devascularization with or without splenectomy for treatment of esophagogastric variceal hemorrhage.Methods The clinical data of 70 patients with a history of esophagogastric variceal hemorrhage from 2009 to 2015 were analyzed.29 cases received spleen preserving paraesophagogastric devascularization and 41 were given portoazygous devascularization plus splenectomy.Results Postoperative portal pressure in spleenpreserving group decreased 14％,that in splenectomy group decreased 23％ (t =2.87,P =0.01).The average blood loss in without splenectomy group was (829 ± 720) ml in contrast to (1 400 ± 1 329) ml in splenectomy group (t =2.311,P =0.024).Postoperative WBC and platelet count in without splenectomy group were lower (t =-5.379,t =-4.924,all P ＜0.01).The postoperative complication rate (24％ vs.39％),and portal venous thrombosis (10％ vs.31％) were all in favour of splenectomy free group (x2 =0.036,P ＜0.05).The 1-and 3-year's recurrent bleeding rate were 4％ and 8％ compared with 4％ and 8％ (all P ＞ 0.05).Conclusion Paraesophagogastric devascularization without splenectomy has less blood loss during the surgery,lower rate of PVT and comparable effect against post-op recurrent bleeding in contrast to devascularization plus splenectomy.

Objective To investigate the complication and risk factors in elderly patients undergoing pancreaticoduodenectomy (PD).Methods From Jan 2005 to Dec 2016,358 consecutive patients undergoing PD were divided into ≥ 70 years group (98 cases) and ＜ 70 years group (260patients).Perioperative complications were compared and risk factors explored between the two groups.Results There were differences in coronary heart disease between the two groups in the preoperative medical comorbidities (P =0.008),and that of albumin,alanine aminotransferase,alkaline phosphatase and creatinine (all P ＜ 0.05).Perioperative mortality was higher in the elderly group than in the younger group (P =0.038),probably related to higher systemic complications (P =0.001).The independent risk factors influencing the postoperative medical complications were preoperative total bilirubin ≥ 171 μmol/L and intraoperative blood loss ≥ 1 000 ml.Conclusions Patients over 70 years old have a comparatively higher perioperative mortality rate after pancreaticoduodenectomy,which were probably derived from a higher postoperative vital organ dysfunction.

OBJECTIVETo identify two novel HLA alleles HLA-B*9536 and B*4612, in an individual.

METHODSDNA was extracted from whole blood by Invitrogen DNA extraction kit. The amplification for HLA-B exons 2-4 of the proband was performed separately with allele group specific primers and the PCR products were directly sequenced for exons 2-4 in both direction.

RESULTSThere were two novel HLA-B alleles in the proband. The sequences of the two alleles have been submitted to GenBank (EU081878 and EU081879). The two alleles have been officially named as B*9536 and B*4612 by the WHO Nomenclature Committee. The sequence of exons 2-4 of HLA-B*9536 showed one nucleotide difference in exon 3 at position 544 (G to A) comparing with the closest allele B*1505, which resulted in an amino acid change from Ala to Thr at codon 158. In the HLA-B*4612 allele, there was one nucleotide change in exon 3 at position 363 (G to A), when compared to the closest allele B*4601, which lead to an amino acid change from Arg to Ser at codon 97.

CONCLUSIONTwo novel HLA-B alleles were identified in one individual and have been officially named by the WHO Nomenclature Committee.

Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; HLA-B Antigens ; genetics ; HLA-B15 Antigen ; Humans ; Male ; Molecular Sequence Data