AIM To study the delayed cardioprotection eff ec ts of pharmacological preconditioning of nitroglycerin (NG) and buprenorphine (B U) used alone or in combination on myocardial ischemia in rats. METHODS Male Wistar rats were randomized to 5 groups. Normal saline (NS) was inf used intravenously with the same volume of NG and BU in Sham group. The anterior mesenteric artery was ligated for 10 min in delayed ischemic preconditioning (D IP) group. BU (1 0 mg?kg -1 ) and NG (0 3 mg?kg -1 ) was administere d intravenously alone or in combination in BU, NG and B+N group respectively. Th e ischemia and reperfusion injury were induced by ligation of left arterior desc ending coronary artery for 30 minutes and followed 2 hours reperfusion at 24 h a fter first operation. Heart rate, blood pressure, ECG ST-segment and arrhythmia s were recorded continuously throughout the whole test. The plasma LDH and CK we re measured. on 30min after ischemia and 2 h after reperfusion. HE and TTC stain ing were performed to determine the myocytes necrosis at the end of test. RESULTS Compared with sham group,DIP group reduced ST-segment during ischemia ( P

Objective To evaluate the effect of propofol (PPF) on stress response and lung injury in rats with traumatic brain injury (TBI).Methods A total of 36 SD rats were randomly (random number)divided into sham group,intralipid group,TBI group,PPF1 h group,PPF 2 h group,PPF 6 h group (n =6 in each group).Fluid percussion brain injury models were used.By intraperitoneal injection,intralipid was administered in intralipid group after sham operations,while propofol 100 mg · kg-1 was given to rats in PPF1 h group,PPF 2 h group and PPF 6 h group 1,2,6 hours following injury,respectively.Nerve motor function were scored at different intervals,serum concentrations of adrenocorticotropic hormone (ACTH),cortisol (COR) and norepinephrine (NE) were measured 12 h after injury.Seventy-two hours later,all rats were sacrificed and brains were harvested for TTC staining,and lungs taken were stained with HE staining for observation under light microscope and electron microscopy.Results Compared with sham and intralipid group,nerve motor function scores were significantly decreased,and serum concentrations of ACTH,COR and NE were increased significantly in rats after injury.Compared with TBI group,the above biomarkers were improved significantly after propofol injection.There were no significant differences in above biomarkersbetweenPPF 1 hand PPF 2 h group (t=-0.816,t=-0.208,t=0.582,P＞0.05).The differences in COR and NE concentrations between PPF 2 h and PPF 6 h group were statistically significant (t =3.018,P =0.013;t =3.662,P =0.004).Light microscopy demonstrated abundant inflammatory cell infiltration and massive thickening of the alveolar walls,Electron microscopy showed Type Ⅱ lung epithelial cell swelling,vacuolar degeneration,osmiophilic lamellar corpuscle emptying in cytoplasm,microvilli protrusions decreases in some cytoplasm in TBI group,and pathological damage was improved significantly in PPF 2 h group.Conclusions Propofol may inhibit stress and protect the lung tissue from damage in TBI rats.

Turnner syndrome is a common sex chromosome disorder. We reported a rare case with Turnner syndrome caused by abnormal number and structure of sex chromosomes. Hereby fluorescence in situ hybridization (FISH) and copy number variation by whole genome low depth sequencing (CNV-seq) were used to clarify the abnormal chromosome. This study provides a diagnostic strategy for clinicians and genetic researchers.

BACKGROUND:Percutaneous vertebroplasty and percutaneous kyphoplasty have become the mainstream clinical methods for the treatment of vertebral compression fractures. However, both of them have several shortcomings such as bone cement leakage, spinal stenosis, nerve compression, pulmonary embolism and other issues. OBJECTIVE:To verify the possibility of bone filing mesh container prepared by polyethylene terephthalate for the treatment of vertebral compression fractures. METHODS:The biological properties of bone filing mesh container were examined according to GB/T16886. After sample aging test, the tensile properties of the aged samples and the fresh prepared samples were compared. The expansion and bone cement leakage were evaluated by injecting bone cement into the bone filing mesh container and measuring the pressure. The initial strength and stiffness of the fresh pig vertebrae with calcium phosphate cement injection or with bone filing mesh container filed with calcium phosphate cement were compared. The in vivo bone tissue growth was periodicaly observed after the lumbar vertebra of 4-month-old pigs was implanted with the bone filing mesh container that was then ful of bone cement. RESULTS AND CONCLUSION: The bone filing mesh container had good biocompatibility. Bone filing mesh containers after 2-year storage had the same tensile strength to the fresh bone filing mesh containers. At ambient conditions, after bone cement injection, bone filing mesh containers could be expanded at 5-10 atm and therefore could play the role of uplift; at 7-10 atm, bone cement could leak out from the bone filing mesh container and enter into the interspace between surrounding bone tissues, thus playing the role of adhesion and fixation. The vertebrae after bone cement injection with or without bone filing mesh containers had the same initial strength and stiffness and exhibited bigger initial strength and stiffness than untreated vertebrae. Thein vivo animal experiments proved that bone filing mesh container had no obvious effect on the vertebrae. These findings indicate that the bone filing mesh container can be used to restore the height and strength of the fractured vertebrae. Moreover, it may eliminate bone cement leakage and therefore increase the surgery safety.

OBJECTIVE: To analyze the clinical features and variants of ABCD1 gene in a Chinese pedigree affected with X-linked adrenoleukodystrophy. METHODS: Clinical data of the proband were collected and analyzed. Potential variant of the ABCD1 gene were analyzed by PCR and Sanger sequencing of the proband, his parents and 100 unrelated healthy individuals. RESULTS: The prominent features of the proband included cerebellar and brainstem lesions, along with increased serum level of very-long chain fatty acids. He was found to harbor a hemizygous c.1509delG (p.L504Sfs*54) variant of the ABCD1 gene, for which his mother was heterozygous. The same variant was not detected in his father and 100 healthy controls. CONCLUSION X-linked adrenoleukodystrophy has a variety of clinical manifestations. Discovery of the c.1509delG (p.L504Sfs*54), as a novel pathogenic variant of the ABCD1 gene, has enabled diagnosis and genetic counseling for this pedigree.
Adrenoleukodystrophy/genetics* ; Asians/genetics* ; China ; Female ; Genetic Testing ; Humans ; Male ; Mutation ; Pedigree

To study the genotype-phenotype and genetic characteristics of Kallmann syndrome. Five patients with Kallmann syndrome were enrolled. Clinical data collection, chromosome karyotyping, whole exome sequencing (WES), and multiplex ligation-dependent probe amplification (MLPA) were used. All the five patients were males, aging from 2 months to 45 years old. Three of the five patients complained cryptorchidism, one complained gonadal dysgenesis, and one complained fasting hyperglycemia. The clinical feature was hypogonadotropic hypogonadism with anosmia, and all karyotype was 46 XY. Magnetic resonance imaging (MRI) showed undeveloped olfactory bulbs and tracts. Kallmann syndrome related gene novel variants were found in all the 5 patients. The hypoplasia of right kidney was found in a patient with c. 1795_1799del (p.Asn599Profs*66) of anosmin 1 (ANOS1) variant. Clinical heterogeneity and incomplete penetrance were seen in a patient with c. 2824A>G (p.Thr942Ala) of chromodomain helicase DNA binding protein 7 (CHD7). Besides, WES indicated a 109 bp-deletion on Xp22.31 (chrX: 8507699-8507804), which was the deletion of exon 10 on ANOS1 gene verified by MLPA. The deletion variant was inherited form his mother, and conformed to X-linked recessive inheritance. Kallmann syndrome is genetic and clinical heterogeneous. WES is helpful for early diagnosis. MLPA and genome copy number variation analysis (CNV) are also recommend if necessary.

Objective:To construct the pET30a-EgG1Y162-2 prokaryotic expression plasmid and induce the expression of EgG1Y162-2 protein, so as to provide a research basis for development of Echinococcus granulosus vaccine. Methods:Using Echinococcus granulosus cDNA as a template, the target gene of EgG1Y162-2 was synthesized by PCR, and after digestion with restriction enzymes EcoRⅠ and Hind Ⅲ, it was connected to the prokaryotic expression vector pET30a to construct the recombinant plasmid pET30a-EgG1Y162-2. The recombinant plasmid was transformed into competent cell BL21 (DE3) and induced by isopropyl β-D-thiogalactoside (IPTG) to express a large number of proteins. The recombinant protein was purified by affinity chromatography. The purification level was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the expression product was identified by Western blotting. Results:The recombinant plasmid pET30a-EgG1Y162-2 was successfully constructed. After inducting expression, the bacterial supernatant and the eluate were both at a relative molecular weight of about 15 × 10 3, and the protein antigen component eluted with 200 mmol/L imidazole was relatively pure. Western blotting results showed that the purified recombinant protein EgG1Y162-2 with His tag could be recognized by His monoclonal antibody. Conclusion:The pET30a-EgG1Y162-2 prokaryotic expression plasmid of Echinococcus granulosus is successfully constructed, and the recombinant protein of EgG1Y162-2 is induced to express, laying a foundation for further study on anti- Echinococcus granulosus vaccine.

Objective:To explore the effects of miRNA-373-3p (miR-373-3p) on the proliferation of nephroblastoma G401 cells through targeted regulation of CD44 expression.Methods:Bioinformatic method was used to predict the possible targeted genes of miR-373-3p based on bioinformatic databases including miRDB, miRanda, PITA and DIANA-microT. G401 cells were taken and transfected with miR-373-3p mimic, mimic negative control, miR-373-3p inhibitor or inhibitor negative control, respectively. Cell proliferation ability was detected by using CCK-8 assay. The number of clones was detected by using clone formation assay. The relative expression level of CD44 mRNA was detected by using real-time fluorescent quantitative polymerase chain reaction (qRT-PCR), and the expression level of CD44 protein was detected by using Western blotting. The dual luciferase gene reporter assay was carried out in HEK-293T cells to vertify the target gene of miR-373-3p.Results:Bioinformatic analysis indicated that CD44 was a targeted gene of miR-373-3p. After 24 h transfection, the proliferation activity of G401 cells in miR-373-3p mimic group was decreased compared with that in mimic negative control group (all P < 0.05). After 48 h transfection, the proliferation activity of tumor cells in miR-373-3p inhibitor group was increased compared with that inhibitor negative control group (all P < 0.05). The formed number of clones in miR-373-3p mimic group was reduced compared with that in the mimic negative control group (55.3±2.5 vs. 90.7±2.9), and the difference was statistically significant ( t = 14.57, P < 0.01). The formed number of clones in miR-373-3p inhibitor group was more than that in inhibitor negative control group (115.0±2.7 vs. 92.0±2.4), and the difference was statistically significant ( t = 8.86, P < 0.01). The dual-luciferase gene reporter assay showed that CD44 was a direct targeted gene of miR-373-3p. The relative expression levels of CD44 mRNA in miR-373-3P mimic and mimic negative control group were 0.62±0.03 and 1.00±0.01, respectively, and the difference was statistically significant ( t = 11.28, P < 0.01). The relative expression levels of CD44 mRNA in miR-373-3p inhibitor and inhibitor negative control group were 1.31±0.02 and 1.00±0.00, respectively, and the difference was statistically significant ( t = 12.65, P < 0.01). The CD44 protein expression was decreased in miR-373-3p mimic group, while increased in miR-373-3p inhibitor group. Conclusion:miR-373-3p can inhibit tumor cell proliferation by targeting CD44 in nephroblastoma.