Objective To analyze the nucleotide sequences of novel allele HLA-B*4608. Methods Genomic DNA of the proband was extracted from whole blood by the commercial DNA extraction kit. The HLA-B exons 1-8 of the proband was amplified and the amplified product was cloned by TOPO TA cloning sequencing kit to split the two alleles apart. Both strands of exons 2, 3 and 4 of chosen colonies were sequenced. Results The sequencing results showed HLA-B alleles of the proband as B*5502 and a new allele as proved by blast search. The sequences of the novel allele have been submitted to Genbank(DQ177521,DQ177522,DQ177523). The new allele is similar to HLA-B*4601 except for two nucleotide substitutions in exon 3: T to C at nucleotide position 538 and G to T at nucleotide position 539. These result in an amino acid substitution at codon 156 from W to L. Conclusion This allele is a novel allele and has been officially named B*4608 by the WHO Nomenclature Committee.

Objective To investigate the significance of serum procalcitonin(PCT)levels in children with respiratory syncytial virus(RSV)pneumonia and sepsis.Methods 43 cases of children with RSV pneumonia were divided into two groups,including 28 cases of non -sepsis and 15 cases of sepsis.30 cases of healthy children were choosed as a control group in the same period.The serum PCT levels of three groups were compared.The 29 cases of RSV pneumonia with other organ damage except lung were divided into non -severe group and severe group.The differences of serum PCT levels between the two groups were compared.Results The non -sepsis group and sepsis group serum PCT levels were (0.882 ±0.184)μg/L and (1.001 ±0.268)μg/L respectively,which were higher than the control group[(0.365 ±0.085)μg/L,(t =2.607,2.854,all P <0.05)].The serum PCT levels of the non -sepsis group and septic group were significant different(t =0.372,P >0.05);The serum PCT level of the severe group was (1.181 ±0.281)μg/L,which was higher than the non -severe group[(0.448 ±0.140)μg/L],the differ-ence was statistically significant(t =2.473,P <0.05).Conclusion The serum PCT of RSV pneumonia children was increased,the increased PCT can be an observed indicators of severe infection and multiple organ dysfunction.

Objective To explore the effect of inferior hypothermy treatment on serum TNF and IL-6 in patients with severe cerebral trauma.Methods 46 patients were randomly divided into two groups:inferior hypothermy group(24 C88e8)and normal group(22 cases).There are the same basic treatments within the two groups,in the inferior hypothermy group we also sive them hypothermy treatment rectal temperature:32～34℃ which need to last for nearly 4～5 days,at the same time we give patients the lyric cocktail.the TNF,IL-6 and GCS grades on the lst and 14th day were tested.Results TNF and IL-6 as compared with normal group are higher than the inferior hypothermy group,the differences between the two groups are of statistical significance(P＜0.01).The difference of GCS grades between the two groups are of stafictical significance(P＜0.05).Conclusion The inferior hypothermy tbempy which inhibits TNF and IL-6 releasing after severe cerebral trauma and the following damages plays a very important role in the cerebral trauma therapy.

Diabetic neuropathic pain(DNP)is one of the most common complications in clinical, which influenced patients’ daily functions greatly, without clear mechanisms and effective methods.P2X3 receptors play a pivotal role in the formation, transmission and conduction of pain under neuropathic pain models, associated with peripheral sensory nerve excitability enhancement.This paper focuses on the establishment of DNP models, and the effects of P2X3 receptors in diabetes mellitus.

Background Retinal ischemia-reperfusion (RIR) injury is a common pathologic change.Its mechanism has not been identified.Objective This study was to investigate the relationship of microRNA-181a (miR-181a) ,tumor necrosis factor-α (TNF-α) and retinal ganglial cells (RGCs) in RIR injury.Methods RIR models were induced in 68 rats,then the rats were randomly divided into control group and RIR groups,including 0hour group,24-hour group and 72-hour group by random number table.Predicted target gene TNF-α was chosen,according to M iRanda,Targetscan and miRBase databases.Immunofluorescent labeling, Western blot and quantitative real-time PCR were used to identify the expression levels of miR-181a,TNF-α and RGCs.Immunofluorescent labeling of RGCs in retinal flat mounts was analyzed for RGCs counts.Results Compared with the control group, RGCs densitiy was obviously decreased in 24-hour and 72-hour RIR groups (P＜0.001).The expression level of mir-181a significantly decreased with reperfusion time in the RIR groups (P＜0.05).Futhermore, the expression level of miR181a was positively correlated with RGCs numbers (r=0.995 ,P=0.005).TNF-α and miR-181a were mainly located in inner layers of retina.As opposed to the changes in RGCs numbers and miR-181a expression,TNF-α in 24-hour group was obviously higher than that of the 0-hour group, though there was no statistical significance in overall correlation analysis.Conclusions In RIR,miR-181a may be involved in regulating RGCs apoptosis.TNF-α may be a target gene of miR-181 a.Interventions within 24 hours after reperfusion might be critical.Further study of miR181 a may help to explore new molecular targets for neuroprotection treatment.

Most colorectal surgeries now are carried out as minimally invasive surgery along with the development and application of laparoscopy.However,the surgical imaging system does not meet the progressing requirement of laparoscopic surgeons.Thanks to the advent of three-dimensional (3D) laparoscopy,a more sophisticated surgical scene with realer images of anatomy is provided to surgeons.It makes identification and manipulation easier,so that to shorten the learning curve.Meanwhile,the 3D laparoscopy is not perfect.But authors believe it will finally be used broadly in colorectal surgery with the improvement of the technology.

We began our teaching reform by following the step of national adjustment of majors frame at the end of last century.We perfected our teaching contingent,created advanced content, composed high quality text book,used multiple teaching methods and ways, paid attention to practical teaching,carried out effective access system and in the end,we firstly combined Meridian and Collateral and Acupuncture,which are the two major courses,into one basic course called Meridian and Acupoint.It covers different levels of students including doctors,postgraduates,undergraduates and seven-year-class students and has the features of abundant and detailed content,creative methods and excellent effects.

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 effects of low frequency electroacupuncture on the P2X3 receptor expres-sion in dorsal root ganglion ( DRG) of rats with type II diabetic neuropathic pain. Methods Part 1:Fifty normal SD rats were randomly divided into normal group (8 rats) and model group (42 rats). The rat model of type II diabetic neuropathic pain was generated by high fat and high sugar diet with a single intraperitoneal injection of streptozotocin ( STZ, 35 mg/kg) . 2 Hz electroacupuncture was administered at ipsilateral acupoints Zusanli and Kunlun for consecutive 7 days. Insulin sensitivity index ( ISI) was measured at 0 w and 5 w, and fasting plasma glucose ( FPG) was measured at 0 w, 5 w, and 7 w. Paw withdrawal threshold (PWT) was measured by mechanical pain threshold, and P2X3 receptor was determined by immunofluorescence. Part 2:Twelve rats with diabetic neuropathic pain (DNP) were divided into EA + vehicle group (6 rats) and EA + αβ?meATP group (6 rats). Rats in the two groups received the same EA treatment as Part 1. Rats in the EA + αβ?meATP group were injected with P2X3 receptor agonistαβ?meATP (0. 6μmol/L, 100μL) into the ventral sur?face of each hind paw every time before EA treatment. Rats in the EA + vehicle group received the same dose of vehicle ( PBS buffer) as a control. Pain threshold of the rats were measured. Results ① Compared with the normal group, the ISI levels of the rats in DNP group was significantly decreased after 5 weeks of the high?fat high?sugar diet (P < 0. 01). Two weeks after STZ injection, the fasting plasma glucose levels in the rats receiving STZ were significantly elevated ( P <0. 01 ) . The type 2 diabetes model was established with a successful rate of 69. 04%. ②PWTs:The PWTs of rats in DNP group were reduced compared with rats in the normal group (P < 0. 01), indicating that the type 2 DNP model was suc?cessfully established. Compared with the PWTs of DNP?controlled rats, the 2 Hz electroacupuncture significantly increased bilateral PWT of rats subjected to DNP from day 3 after treatment (P < 0. 01). P2X3 receptor agonistαβ?meATP greatly reduced bilateral PWT of EA?treated DNP rats compared with that of the EA + vehicle group (P < 0. 01). ③The immu?nofluorescence essay showed that P2X3 receptor expression in bilateral L5 DRGs in the DNP group was significantly in?creased as compared with that in the normal group ( P < 0. 01 ) . The increases were inhibited by 2 Hz EA in L5 DRGs compared with the DNP group (P < 0. 01). Conclusions 2Hz electroacupuncture can effectively treat the type II diabet?ic neuropathic pain by decreasing the expression of L5 DRG P2X3 receptors in rats.

Objective To investigate the risk factors of severe hand,foot and mouth disease (HFMD).Methods 175 severe cases of HFMD and 183 mild cases of HFMD in the same period were randomly selected.Single factor analysis was first performed between severe and mild cases on age,gender,residence,symptoms,signs and laboratory examinations,etc,to screen out the related risk factors which P value ＜ 0.05.Then,binary logistic regression analysis was carried out to determine risk factors most related to severe HFMD.Finally,receiver operating characteristic curve (ROC) analysis was performed on severe HFMD related risk factors.Results Single factor analysis showed that there were obvious differences between children with mild HFMD and those with severe HFMD in the factors like difficulty in breathing,walking instability,vomiting,limb shaking,disturbance of consciousness,convulsions,cold sweat and weakness,thermal process,the degree of fever,pulmonary rales,heart rate,serum EV71 antibody,circulatory failure,leukocyte count,platelet count,neutrophil ratio,CRP,blood glucose,etc (x2 =15.236,19.819,33.823,52.670,12.984,10.180,29.318,52.932,34.544,14.615,46.633,31.407 and 5.303,t =3.184,3.144,2.256,2.244 and 2.828,,all P ＜0.05).Binary logistic regression analysis showed that the thermal process,startle tremor or limb jitter,serum EV71 antibody,vomiting,fever,neutrophil ratio were the related risk factors of severe HFMD (B value =2.605,2.129,1.409,1.185,0.841 and 0.103,all P ＜ 0.05).ROC analysis showed that the areas under the curve of the predicted probability and thermal process were larger than any other risk factors [(95％ CI (0.888 ～ 0.961) and (0.818 ～ 0.920)],and thus had better diagnostic values.Conclusion Children under 3 years old were the high risk population of HFMD.Such clinical symptoms as persistent high fever,vomiting,startle tremor orlimb jitter,EV71 antibody in serum and increasing neutrophil ratio were risk factors for severe HFMD.The predicted probability had more diagnostic value than any other risk factors.