Objective To analyze the blood screening results after adjustment of critical value 40 to 50 U/L and to observe the effect of reducing blood scrap rate and to discuss the correlation between ALT and HBV,HCV infection.Methods We screened 2656 blood donors (ALT >40 U/L)by serological and nucleic acid amplification testing(NAT)in Qingdao blood center from 2013 to 2014,and conducted the correlation analysis by chi square test.Results 1 771 cases (66.68%)were ALT 40-50 U/L,including 6 cases of HBsAg ELISA (+),2 cases NAT (+),4 cases NAT(-).In the 8 cases of anti-HCV ELISA (+)samples,4 cases NAT (+),3 cases NAT (-),1 case with positive TP without NAT result.In 885 blood donors with ALT>50 U/L,5 cases were HBsAg-reactive,7 cases were anti-HCV-reactive,and 873 cases were negative.Related statistics showed that there was no signifi-cant difference between ALT and HBV infection (P <0.05),but significant difference was found between ALT and HCV infection (P >0.05).Conclusion The proportion of blood donors with ALT 40-50 U/L is much higher than that with ALT >50 U/L do-nors.Adjustment of the critical value greatly reduces blood scrap rate.Elevated ALT is associated with the infection of HBV but not with HCV.

Objective To observe the influence of Xiaoshuan enteric-coated Capsule (XSECC) on neuronal damage in cortex of cerebral hypo-perfusion rats. Methods Rats were divided into a sham group (12), a model group (17), a XSECC large dose group (15), a medium dose group (15) and a low dose group (15) by a random number table. The cerebral hypo-perfusion model was produced by permanent bilateral common carotid artery ligation (2VO). The mixed suspension of XSECC was given orally to rats in the XSECC large dose group (420 mg/kg), the medium dose group (140 mg/kg) and the low dose group (47 mg/kg) once each day for 40 days from the beginning of two hour after ischemia. The expressions of NeuN and Caspase-3 were observed by immunofluorescence staining at 40 days after ischemia. The content of GSH-PX,SOD and MDA were measured by biochemical assay. Results Compared to the model group, the expressions of NeuN (8 716.86 ± 2 539.93, 9 549.31 ± 1 663.26 vs. 7 297.05 ± 1 932.49) were significantly increased in the XSECC large dose group and the medium dose group (P<0.05 or P<0.01);The content of GSH-PX (7.37 ± 1.08 U/mg, 7.77 ± 3.26 U/mg vs. 3.67 ± 2.52 U/mg) was increased in the XSECC large dose group and the medium dose group(P<0.05); The expressions of Caspase-3 (11.65 ± 2.68, 14.05 ± 4.55, 12.60 ± 4.56 vs. 16.80 ± 5.41) were obviously decreased in the XSECC large dose group, the medium dose group and the low dose group, compared with the model group (P<0.05 or P<0.01);The content of MDA (1.44 ± 0.40 nmol/mg, 1.96 ± 1.13 nmol/mg, 2.12 ± 1.19 nmol/mg vs. 3.19 ± 0.98 nmol/mg )was decreased in the XSECC large dose group, the medium dose group and the low dose group when compared with the model group (P<0.05 or P<0.01);The content of SOD (555.61 ± 92.45 U/mg, 607.90 ± 228.45 U/mg, 515.98 ± 184.01 U/mg vs. 348.12 ± 108.84 U/mg) was increased in the XSECC large dose group, the medium dose group and the low dose group when compared with the model group (P<0.05 or P<0.01). Conclusion XSECC could protect injured neurons, which is related to the improvement of free radical metabolism.

We have assessed the relationships between immune trait (antibody titers of Sheep red blood cell, SRBC; Avian influenza, AI; Newcastle disease, ND) and varieties of MHC B-LBHII Gene in local chicken breeds (Wenshang Barred chicken, LH; Laiwu Black chicken, LWH; and Jining Bairi chicken, BR). We selected 300 chickens randomly from the three indigenous chicken populations. The variations of MHC B-L BII gene were detected by directly DNA sequencing and polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). The results indicated that there were about 19-22 nucleotide mutations in the three local breeds, which could affect 16-18 amino acid variations. Another results indicated that there was significantly relationship between seven to eight SNPs of the MHC B-LBII region and some immune traits (P < 0.05 or P < 0.01). Both locus G97A and locus T138A were found in the three species, which were significantly related to the antibodies of SRBC, ND and AI antibody titers (P < 0.05). Among them, the locus G97A was significantly associated with ND antibody titers (P < 0.05) in BR chicken, with SRBC antibody titers (P < 0.05) in LWH chicken, and with H9 antibody titers (P < 0.05) in LH chicken. Furthermore, locus T138A was significantly associated with H9 antibody titers in BR and LH chickens (P < 0.05). All those results suggest relationships among the different varieties of MHC B-LBII and immune traits in the three local breeds.
Animals ; Base Sequence ; Breeding ; Chickens ; genetics ; immunology ; Major Histocompatibility Complex ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Polymorphism, Single-Stranded Conformational ; Sequence Analysis, DNA

Objective To analyze the clinical diagnosis and treatment data of 20 children with hypophosphatemic rickets (HR) in order to improve the clinical diagnosis and treatment of HR.Methods The retrospective analysis of clinical data of 20 cases with HR who were hospitalized at Children's Hospital of Nanjing Medical University from May,2010 to April,2016 was performed to summarize the clinical characteristics.All patients were analyzed for the phosphate regulating gene with homologies to endopeptidase on the X chromosome(PHEX) gene by direct sequencing.If no mutations were detected,multiplex ligation-dependent probe amplification analysis was performed.Results All of the 20 cases with HR showed different degrees of growth retardation and typical X-ray rickets.After treatment,the clinical features were improved.Height standard deviation score (HSDS) was improved significantly with longer treatment time,and the difference was statistically significant(P =0.027).There was a correlation between the blood phosphorus fluctuation and secondary hyperparathyroidism(P ＜ 0.05).Nineteen cases had PHEX gene mutations.Truncating mutations was the most frequent mutation type,and 4 new mutations were found.Conclusions Clinical characteristics,laboratory test results and X-ray examination are important clinical index for the diagnosis of HR,and PHEX gene test can be used as an important auxiliary diagnostic tool.Early diagnosis and treatment can significantly improve the clinical manifestations of the patients.

Objective To investigate the impact of iron overload on apoptosis in primary mouse liver cells via a synchronous separation technology.Methods Hepatocyte(HC),liver sinusoidal endothelial cell(LSEC),and Kupffer cell(KC)were isolated and purified with collagenase,percoll density gradient centrifugation,and CD146 magnetic beads.Cell types were identified using flow cytometry and immunofluorescence staining.Cells of different types were cultured in vitro,and an iron overload model was established by treating the mice with 0,25,50 and 100 μmol/L ferric ammonium citrate(FAC)for 24 h.The iron content was quantified using Prussian blue staining,while cell viability and mitochondrial membrane potential were assessed by flow cytometry.Results The synchronous separation technology of primary liver cells exhibited stable efficiency.The yield of HC was(4.0±0.5)×107 cells per mouse,exhibiting an effective survival rate of(76.33±0.67)％.The yield of LSEC was(5.0±1.0)×106 cells per mouse,with a survival rate of(93.63±0.25)％and a purity level of(93.40±0.46)％.The yield of KC was(1.5±0.5)×106 cells per mouse while a high survival rate of(98.33±0.12)％and a purity level of(88.30±2.02)％were maintained.The obtained cells were large in number,with good vitality and high purity,which could meet the requirements of subsequent experiments.Treatment with FAC significantly elevated iron contents in different types of cells when compared with the control group.Upon stimulation of FAC,the survival rate of HC decreased from(73.97±5.54)％to(54.10±1.68)％,the mean fluorescence intensity of JC-1 aggregates decreased from 326.33±30.37 to 155.00±6.56,JC-1 monomer increased from 1700.00±1 44.04 to 3713.33±81.82.The survival rate of LSEC decreased from(90.60±1.74)％to(78.03±2.15)％,the mean fluorescence intensity of JC-1 aggregates decreased from 502.33±5.51 to 372.33±4.04,and JC-1 monomer increased from 750.00±67.51 to 1340.00±36.39.The survival rate of KC decreased from(94.23±1.44)％to(88.37±1.56)％,the mean fluorescence intensity of JC-1 aggregates decreased from 652.67±25.66 to 478.00±12.49,and JC-1 monomer increased from 1984.33±80.65 to 3062.33±245.20.Conclusion A robust and reliable simultaneous isolation technique of primary mouse HC,LSEC,and KC has been established.Moreover,our finding demonstrates that iron overload significantly enhances apoptosis levels in HC,LSEC and KC.

【Objective】 To compare the detection performance of Cobas s201, a minipool(MP) nucleic acid test(NAT) system, and Panther, a individual donation(ID) NAT system, in blood donor screening. 【Methods】 NAT was conducted on 126 359 blood samples, and initially reactive (IR) samples were either discriminated or resolved by ID testing.The non-discriminated reactive (NDR) samples implicated in Panther sysytem were subjected to ID-NAT by Cobas s201. Some non-repeatable reactivet(NRR) and repeatable reactive (RR) samples implicated in Cobas s201 system were subjected to ID-NAT by Panther. 【Results】 61 MP-IR cases were implicated in a total of 85 128 samples that detected by Cobas 201, and 29(0.34‰) were RR after resolved by ID testing. 74(1.79‰)IR samples were implicated in 41 231 samples that detected by Panther, and 22 (29.73%) were DR-HBV after discriminatory test. Among the NDR 28 samples detected by Panther multiplex system, 7 were positive by Cobas s201 single sample (PP1) whereas non-reactive in simulated MPs of six by Cobas 201.In 28 RR samples resolved by Cobas 201, 24 positive and 4 negative samples were retested by Panther. Among the 11 samples presenting inconsistent retest results by Panther and Cobas 201, 10 were anti-HBc positive, carrying low viral load HBV. 【Conclusion】 The NAT-yield by Panther was significantly higher than that by Cobas s201. Some samples with negative discriminatory results were OBI, and it is necessary to further track and verify the unidentified samples. Cobas s201 is more suitable for a wide array of MP-NAT testing while Panther sample loading, which is flexible and easy to operate, is more suitable for ID-NAT with medium sample size.

【Objective】 To study and analyze the serological and viral charactereristics of hepatitis B virus(HBV) infection in voluntary blood donors in Qingdao. 【Methods】 315 520 blood samples of voluntary blood donors were screened by ELISA combined with nucleic acid testing (NAT). All HBsAg-/HBV DNA+ samples were subjected to high-precision viral load detection and five serological markers of HBV. The sequence of HBV S gene was detected by PCR direct sequencing, and virus genotypes and amino acid mutations were analyzed. 【Results】 A total of 604(0.20%)HBV ELISA or NAT reactive samples were detected: HBsAg+ /HBV DNA- in 307(0.10%) cases, HBsAg-/HBV DNA+ in 138(0.04%) and HBsAg+ /HBV DNA+ in 157(0.05%). Among the 138 HBsAg-/HBV DNA+ donors, 118(85.5%) carried anti-HBc, and 45 (32.61%) carried sole anti-HBc and 5 (3.62%) carried both HBsAg and anti-HBc. In viral load detection, 64 were quantitatively negative and 74 were quantitatively positive, of which 42 were HBV DNA <20 IU/mL and 32 > 20 IU/mL. 13 HBsAg-/HBV DNA+ samples were successfully amplified and sequenced, and 5 were genotype B, presenting a total of 17 amino acid mutations without any deletion or insertion, and 8 were genotype C, presenting a total of 41 amino acid mutations and 2 amino acid deletions. 【Conclusion】 NAT, in combination of ELISA, provides additional safety in detecting potentially infectious HBV during the window period and occult HBV infection (OBI). The viral load was low in OBI infected donors, and anti-HBc+ was the main manifestation.The dominating HBV genotypes are genotype B and C, suggesting HBsAg amino acid mutations may be related to the formation of OBI.

OBJECTIVE: To investigate the application value of whole exome sequencing technology in fetuses with congenital structural abnormalities. METHODS: The chromosomal abnormalities of 1147 families were analyzed. According to the follow-up results, the data of fetuses with new phenotypes in late pregnancy or after birth were reanalyzed. Subgroups were divided according to the organs involved and whether single malformation or not. The gene regulatory network map was drawn by using string database and Cytoscape software. Fisher exact probability method was used to compare the difference of the diagnostic rate of pathogenic genes among the groups. RESULTS: A total of 160 fetal cases received positive molecular diagnosed, involving 178 variant sites of 125 pathogenic genes, including 8 cases (4.9%, 8/163) by data reanalysis, and the overall positive diagnosis rate was 13.9%. Diagnostic rate was highest in the group of skeletal malformation (31.5%, 39/124) and lowest in that with thoracic malformation (0, 0/32). The gene clusters of fetal edema and intrauterine growth restriction were independent, and were not associated with the major structural malformations. The probability of each parent carrying the same recessive gene variant was 0.03 (39/1146) and 0.08 (4/53) with positive family history. CONCLUSION For fetuses with congenital structural abnormalities that are negative for conventional genetic tests, 13.9% of phenotypic associated pathogenic/likely pathogenic genetic variants can be detected by whole exome sequencing technology. Its application value for prenatal diagnosis varies in fetus with different organs involved. Reanalysis of sequencing data for cases with new phenotypes in late pregnancy or after birth can further improve the molecular diagnosis rate. Further investigations are needed to explore the related genetic mechanisms.
Female ; Fetal Diseases ; Fetus/diagnostic imaging* ; Humans ; Pregnancy ; Prenatal Diagnosis ; Technology ; Ultrasonography, Prenatal ; Whole Exome Sequencing

BACKGROUND:Due to the sudden release and the rapid removal by proteases,platelet-rich plasma hydrogel leads to shorter residence times of growth factors at the wound site.In recent years,researchers have focused on the use of hydrogels to encapsulate platelet-rich plasma in order to improve the deficiency of platelet-rich plasma hydrogels. OBJECTIVE:To prepare self-assembled polypeptide-platelet-rich plasma hydrogel and to explore its effects on the release of bioactive factors of platelet-rich plasma. METHODS:The self-assembled polypeptide was synthesized by the solid-phase synthesis method,and the solution was prepared by D-PBS.Hydrogels were prepared by mixing different volumes of polypeptide solutions with platelet-rich plasma and calcium chloride/thrombin solutions,so that the final mass fraction of polypeptides in the system was 0.1%,0.3%,and 0.5%,respectively.The hydrogel state was observed,and the release of growth factors in platelet-rich plasma was detected in vitro.The polypeptide self-assembly was stimulated by mixing 1%polypeptide solution with 1%human serum albumin solution,so that the final mass fraction of the polypeptide was 0.1%,0.3%,and 0.5%,respectively.The flow state of the liquid was observed,and the rheological mechanical properties of the self-assembled polypeptide were tested.The microstructure of polypeptide(mass fraction of 0.1%and 0.001%)-human serum albumin solution was observed by scanning electron microscope and transmission electron microscope. RESULTS AND CONCLUSION:(1)Hydrogels could be formed between different volumes of polypeptide solution and platelet-rich plasma.Compared with platelet-rich plasma hydrogels,0.1%and 0.3%polypeptide-platelet-rich plasma hydrogels could alleviate the sudden release of epidermal growth factor and vascular endothelial growth factor,and extend the release time to 48 hours.(2)After the addition of human serum albumin,the 0.1%polypeptide group still exhibited a flowing liquid,the 0.3%polypeptide group was semi-liquid,and the 0.5%polypeptide group stimulated self-assembly to form hydrogel.It was determined that human serum albumin in platelet-rich plasma could stimulate the self-assembly of polypeptides.With the increase of the mass fraction of the polypeptide,the higher the storage modulus of the self-assembled polypeptide,the easier it was to form glue.(3)Transmission electron microscopy exhibited that the polypeptide nanofibers were short and disordered before the addition of human serum albumin.After the addition of human serum albumin,the polypeptide nanofibers became significantly longer and cross-linked into bundles,forming a dense fiber network structure.Under a scanning electron microscope,the polypeptides displayed a disordered lamellar structure before adding human serum albumin.After the addition of human serum albumin,the polypeptides self-assembled into cross-linked and densely arranged porous structures.(4)In conclusion,the novel polypeptide can self-assemble triggered by platelet-rich plasma and the self-assembly effect can be accurately adjusted according to the ratio of human serum albumin to polypeptide.This polypeptide has a sustained release effect on the growth factors of platelet-rich plasma,which can be used as a new biomaterial for tissue repair.

Objective To observe the effect of TCM compound of resolving stagnation for tranquilization ( RST) on neurotransmitters and brain derived neurotrophic factors ( BNDF ) in depression rat model induced by chronic unpredictable stress ( CUS) , and to reveal the mechanism of RST on the treatment of depression.Methods Altogether 60 male SD rats were randomly divided into normal group, model group, high-, mid-and low-dose RST groups at dosage of 8.2, 16.3, 32.7 g/kg, and fluoxetine group with dose of 10 mg/kg.The rats of four therapy groups were administered intragastrically drugs with undergoing CUS for consecutive thirty-five days.The behavioral changes of rats were observed by open field test and novelty-suppressed feeding-test.The content of dopamine ( DA) , 5-hydroxeytryptamine(5-HT) , 5-hydroxyindole acetic acid (5-HIAA) and norepi-nephrine( NE) in hippocampus and cortex were determined by HPLC.And the expression of BDNF in hippocampus was measured by Western blot.Results Compared with the normal group, the rats ’ weight increased slowlier and the spontaneous activity decreased in model group; the contents of 5-HT and NE in cortex and the content of 5-HT, 5-HIAA and NE in hippocampus significantly reduced ( P<0.05 ).Compared with the model group, the spontaneous activity significantly increased in all therapy groups;the contents of 5-HT and NE in cortex dramatically increased in fluoxetine group and mid-, high-dose group ( P<0.05 );the contents of 5-HT and 5-HIAA in hippocampus significantly increased in all three RST groups ( P<0.05 ); the content of NE in hippocampus significantly increased in low-and high-dose group ( P<0.05 ).The expression of BDNF in hippocampus significantly increased in fluoxetine group and high-dose RST group ( P<0.05).Conclusion The anti-depression mechanism of TCM compound formulated by principle of resolving stagnation for tranquilization may be related to the effects of increasing neurotransmitters and up-regulation of BDNF.