Objective To evaluate the accuracy and clinical practicality of DNA chip in comparison with serology in typing of human leukocyte antigen A (HLA-A) in Han's individuals of donor-recipients of transplantation. Methods 120 peripheral blood samples were obtained from donor-recipients of transplantation. Each sample was divided into two parts and HLA-A antigens were identified by DNA chip in one part and by serology in another. Samples in which the HLA-A typing results by these 2 methods were discordant were verified by polymerase chain reaction with sequence specific primers (PCR-SSP). Accuracy and clinical practicability of both methods were compared according to the typing results. Results Serological typing for HLA-A took 3 h, while DNA chip typing 4. 5-5 h. 112 samples have been typed successfully. Typing results were same in 91 samples and discordant in 21 cases. The verified results showed that DNA chip made 2 incorrect typing and the error rate was 2%. Meanwhile, serology made 19 mistakes, consisting of 5 antigens being incorrectly interpreted and 14 "blanks" turning out to be definable alleles. The discrepancy rate was 17 %. Conclusions DNA chip typing for HLA-A is suitable for clinical application in Chinese Han's population with a greater precision than serology. It may replace the serology in future after being improved and perfected.

Objectives To explore the effect of holder pasteurization, frozen storage time and thawing methods on macronutrients and energy content of donor human milk, and to provide theoretical basis for the rational use of breast milk. Methods Thirty-three samples of donor human milk were collected and an aliquot of each sample was analyzed before and after pasteurization. The remaining milk after pasteurization was split into 9 aliquot , and frozen at -20 ℃. After 30, 60, and 90 days, the milk was thawed by three different methods of room tempe-rature, 4 ℃ refrigeration, and 37 ℃ water bath, respectively. The nutrient components of each aliquot were analyzed and compared. Results We observed a mild reduction in fat and energy content after pasteurization (P <0.05). A significant decrease of fat, protein and energy content with the prolonged storage time was observed (P <0.01), and during the whole process (pasteurization + frozen storage), the decrease of fat, protein and energy content was 36.6%, 32.6%and 22.6%, respectively. The protein was influenced mostly by different thawing methods and the content of protein reached highest while thawed at 4 ℃ refrigeration. Conclusions Holder pasteurization and frozen storage at-20℃significantly reduce fat, protein and energy content of donor human milk. The donor milk should be used as quickly as possible when applied for preterm infants and thawing at 4 ℃ refrigeration is recommended before delivery to newborn infants.

To determine the effects of Von Hippel-Lindau (VHL) on the invasion and migration of glioma U251 cells. Methods:U251 GBM cells were transfected using VHL expression plasmid. Real-time polymerase chain reaction was conducted to de-tect VHL mRNA expression after transfection. Western blot assay was used to measure protein (VHL, MMP-2, and MMP-9) expres-sion. Tumor invasion and migration were examined by the Transwell and wound-healing experimental methods after VHL up-regula-tion. The intracranial model of nude mouse was developed using U251 cells transfected by VHL expression plasmid, and immunohisto-chemical staining was used to measure protein (VHL, MMP-2, and MMP-9) expression in the tissue sections. Results: In the U251 cells transfected by VHL expression plasmid, the expression of VHL mRNA and VHL proteins increased, and the expression of MMP-2 and MMP-9 protein decreased. Meanwhile, the invasion and migration of glioma U251 cells were also inhibited. Immunohistochemical staining results showed that the expression of MMP-2 and MMP-9 proteins decreased, and the VHL protein expression increased after transfection. Conclusion:VHL can inhibit the invasion and migration of glioma U251 cells. Thus, VHL gene can be used as a target for the gene therapy of gliomas.

AIM:To investigate the effect of RWDD3 gene silencing on the biological characteristics of human glioma U251 cells.METHODS: A lentiviral vector expressing RWDD3 shRNA was constructed and transfeeted into the U251 cells.The expression of RWDD3 at mRNA and protein levels was detected by real-time PCR and Western blot , re-spectively .The cell activity was determined by MTT assay .The colony formation ability was detected by the colony forma-tion assay .The cell proliferation ability was detected by BrdU incorporation assay .The cell invasion and migration were evaluated by Transwell assay .Flow cytometry was used to monitor the changes of cell cycle distribution and apoptosis .RE-SULTS:Recombinant lentivirus was successfully transfected into U 251 cells.Compared with the cells transfected with the scrambled shRNA and control cells, the cell activity, colony formation ability, and the invasive and migratory activities were inhibited, the cell cycle was arrested in G 0/G1 phase, and the apoptosis was increased in the U 251 cells transfected with RWDD3 shRNA ( P <0.05 ) .CONCLUSION: RWDD3 plays a vital role in proliferation and invasion of glioma cells.It may serve as a potential target of gene therapy for glioma .

Objective:To investigate the effects of iron deficiency in early life on neurobehavioral manifestations in CDH2 genetic mutation rats, in order to explore interaction between iron deficiency and CDH2 gene mutation in autism-like behavior.Methods:The 16 female SD rats with established CDH2 genetic mutations were randomly divided into 8 rats as iron deficiency group and 8 rats as normal diet control group.After mating, rats were fed low-iron diets during pregnancy and lactation or standard diets as control.The offsprings were randomly divided into 27 rats as iron deficiency group and 27 rats as control group.The voice communication ability of offspring was studied through ultrasonic vocalizations.The data and videos of open-field test, three-chamber social interaction test were recorded by animal behavioral video analysis system(Smart3.0). The characteristics of behavior changes were analyzed by statistical methods.Results:Total number of calls in rats aged 10 day in iron deficiency group[(755.67±161.86)times]were significantly lower than that in the control group[(1461.89±166.57)times]( P<0.05). Total calling duration[(41.77±16.17)s]were significantly lower than that in the control group[(86.22±10.07)s]( P<0.05). There is a certain synergistic effect of CDH2 genetic mutation and iron deficiency on the reduction of total calls in rats aged 6 and 8 day( P<0.05). The total distance, distance in zone-periphery, mean speed in zone-total and mean speed in zone-periphery of rats in iron deficiency group were all higher than that in the control group( P<0.05); the numbers of standing and rearing were reduced( P<0.05). In the second stage of the three-chamber test, the rats in the iron deficiency group had shorter communication time with the new stranger 2 and longer interaction time with the old stranger 1 compared with the control, showing a weakening trend of social ability, but the difference was not statistically significant( P>0.05). Conclusion:Autism-like behavioral changes occurred in rats with iron deficiency in early life and CDH2 genetic mutation, and there is a certain degree of synergy.Iron deficiency and CDH2 mutation may increase the risk of autism spectrum disorder.

Objective To investigate the effects of low GI cereals on metabolomics and pregnancy outcomes in women with gestational diabetes mellitus (GDM),in order to explore the preventive and therapeutic mechanisms and provide the basis for nutritional interventions.Metbods Pregnant women with gestational diabetes were assigned to the treatment group (n=31),using low GI grains 12 weeks for nutrition intervention and the control group (n =31) according to the random digital table method;30 healthy pregnant women were enrolled as normal control group.At 36th gestational week serum was analyzed by 1H-NMR metabolomics approach.Pregnancy outcomes were gathered for statistics after delivery.Comparison among groups and related influencing factors analysis were conducted.Results After nutritional intervention for 12 weeks,there were statistically significant differences in 15 potential biomarkers associated with gestational diabetes between the treatment group and the control group (P＜0.05),and no statistically significant difference between the treatment group and the normal control group (P＞0.05),that was the pregnant women in the treatment group were close to normal pregnant women.Cesarean rate,gestational weight gain,glycosylated hemoglobin during delivery,fasting insulin and newborn birth weight were significantly lower in the treatment group than in the control group.Multiple linear regression analysis showed that the GI of diet,fasting insulin and blood glucose were influencing factors for metabolomics in women with GDM.Conclusions Using low GI cereals intervention treatments,the pregnancy outcomes of GDM are improved distinctly with the possible mechanisms as adjusting the related biomarkers.Our study provides evidences for further exploring etiology and the therapeutic mechanisms of GDM,and individualized medical nutrition treatment strategy.

Objective To evaluate the short-term effect of Pipeline embolization device (PED)for the treatment of unruptured wide-necked intracranial aneurysms. Methods From October 2015 to September 2016,15 consecutive patients with unruptured wide-necked intracranial aneurysm (aneurysm neck and aneurysm body ratio ≥0. 5)treated with PED at the Department of Neurosurgery,the Second Affiliated Hospital of Nanchang University were enrolled retrospectively. Their clinical and imaging data were analyzed. Kamran scale was used to evaluate the embolization rate of aneurysms and the changes of the parent arteries. DSA examination was performed again at 6 -12 months after operation. Results Fifteen PED were implanted in 15 patients with unruptured wide-necked intracranial aneurysms,including 13ophthalmic artery aneurysms,1 posterior communicating artery aneurysm,and 1 cavernous sinus aneurysm. The technical success rate was 100% . Immediately after PED implantation,Karman rating of 15 cases were aneurysm grade 2 embolization,parent artery grade A (grade 2a). DSA examination was performed again at 6 - 12 months after operation showed that 14 patients were aneurysm grade 4,parent artery was grade A (grade 4a). One patient (ophthalmic artery aneurysm)underwent the second DSA examinations at 6 and 12 months after operation showed that the residual development of aneurysms. The aneurysm embolization was grade 3, and the parent artery was grade A (grade 3a). No branch artery occlusion was observed. Non of them had neurological deficit. The modified Rankin scale score was 0 in all 15 patients. Conclusion The use of PED in the treatment of unruptured wide-necked intracranial aneurysms has a higher occlusion rate. Its long-term effect still needs further follow-up.

Objective:To explore the characteristics of gut microbiota in the preoperative, short-term postoperative and long-term postoperative period at (15.61±4.51) months in children with ventricular septal defect (VSD) of congenital heart disease (CHD) treated with cardiopulmonary bypass (CPB).Methods:A prospective study was conducted.In Guangzhou Women and Children′s Medical Center, 13 patients with VSD who were scheduled for CPB and additional 10 age- and gender-matched healthy infants as pre-CPB control group from January 2021 to January 2022 were enrolled.Fecal samples were collected at pre- and early post-CPB.Meanwhile, 18 gender- and CHD diagnosis and operation-matched patients at (15.61±4.51) months after CPB and 8 healthy age- and gender-matched children as long-term control group after CPB were also enrolled, and fecal samples were collected.16S rRNA sequencing of fecal samples from all subjects were performed and comparing the differences in gut microbiota between two groups via comparing alpha and beta diversity, parameter test or nonparametric test, and LEfSe analysis.Results:Compared with those of pre-CPB control group, there was a significant difference in the composition of gut microbiota in the preoperative period of VSD children, with significantly increased abundances of Enterobacteriaceae and Shigella, and decreased abundance of Bifidobacterium (all P<0.05). The diversity of gut microbiota was comparable in VSD children before CPB and in the short period time after CPB (all P>0.05), except for the abundances of Clostridium and Streptococcus (all P<0.05), and there was no significant difference in the relative abundances of other highly abundant gut bacteria between the two periods (all P>0.05). Compared with that in VSD children in the short period time after CPB, the abundances of short-chain fatty acids-producing microbes were significantly higher at (15.61±4.51) months postoperatively (all P<0.05), and the gut bacteria profile was similar to that of the long-term control group after CPB (all P>0.05). Conclusions:Gut microbiota imbalance exists in VSD children before CPB.The gut microbiota profile is not influenced by CPB, which returns normal at (15.61±4.51) months postoperatively.

Objective To study the effect of exsomes in SH-SY5Y cells after hypoxic ischemia/reperfusion injury.Methods Human umbilical vein endothelial cell hypoxic ischemia/reperfusion (HUVEC I/R) injury models were established,and the exosomes derived from HUVEC I/R were extracted and identified.SH-SY5Y cell hypoxic ischemia/reperfusion injury models (SH-SY5Y I/R) were established,and cells from SH-SY5Y I/R were divided into control group and exosomes-treatedgroup.The proliferation of SH-SY5Y cells was evaluated by CCK-8 assay 24,48and 72 h after cell inoculation.Transwell assay and wound-healing assay were used to examine the invasion and migration.Hochest33258 staining and Flow cytometry were used to monitor the changes of cell cycle and apoptosis.Expressions of Caspase-3,Bax and Bcl-2 were measured by real-time fluorescence quantificative-PCR and Western blotting.Results As compared with those in the control group,the proliferation abilities of SH-SY5Y cells in exosomes-treated group were significantly promoted (48 h:0.70±0.05 vs.0.94±0.08;72 h:0.83±0.05 vs.1.02±0.06),the cell cycle rate of S phase was significantly increased (14.39％±4.11％ vs.20.54％±3.46％),and G0/G1 phase was statistically decreased (71.26％± 5.24％ vs.66.87％±4.23％,P＜0.05).What's more,cell invasive was significantly promoted (44.00±6.56 vs.70.67±6.11),and relative wound injury area was significantly reduced in the exosomes treated group (0.61±0.07 vs.0.52±0.10);significant differences were noted between the two groups (P＜0.05).The mRNA expressions of Bax and Caspase-3 were significantly decreased and the mRNA expressions of Bcl-2 was significantly increased in the exosomes-treated group as compared with those in the control group (P＜0.05).Conclusion HUVEC I/R-derived exosomes play neuro-protective role in human SH-SY5Y cells after hypoxic I/R injury.