Cell Physiological Phenomena ; Electromagnetic Fields

Objective To prepare the anti-hUART1 polyclonal antibody and investigate the subcellular localization of hURAT1 protein. Methods The full-length hURAT1 gene was obtained by RT-PCR and inserted into the fusion expression vector pEGFP-N3,the predicated antigen epitope was cloned into GST fusion protein expression plasmid pGEX-5X-1, transformed into E. coli BL21 cells for expressing recombinant GST-hURAT1 protein induced by IPTG. The purified hURAT1-GST fusion protein was employed to immunize rabbit for preparing the polyclonal antibody. The expression of hURAT1 was analyzed by western-blot and immunohistochemistry in human kidney. pEGFP-hURAT1 was transfected into the LLC-PK1 cell in order to observe the subcellular localization of the gene using confocal microscopy. Results Specific anti-hURAT1 rabbit polyclonal antibody was obtained, and both Western-blot and immunohistochemistry showed that hURAT1 was expressed in the human kidney brush border,localized in the apical membrane of the LLC-PK1 cell. Conclusions hURAT1 protein was a membrane protein located in renal proximal tubule, which could be detected in the apical membrane. The anti-hURAT1 polyclonal antibody could be used for studying the physiological function of hURAT1 and its pathology.

Apoptosis ; radiation effects ; Cell Cycle ; radiation effects ; Cells, Cultured ; Electromagnetic Fields ; Gene Expression ; radiation effects ; Mutation ; radiation effects ; Recombination, Genetic ; radiation effects ; Yeasts ; cytology ; genetics ; radiation effects

Aged ; Female ; Humans ; Immediate Dental Implant Loading ; methods ; Jaw, Edentulous ; diagnostic imaging ; rehabilitation ; surgery ; Male ; Mandible ; diagnostic imaging ; surgery ; Maxilla ; diagnostic imaging ; surgery ; Middle Aged ; Radiography, Panoramic ; Surgery, Computer-Assisted ; methods

Early hematoma enlargement in intracerebral hemorrhage seriously influences the prognosis of patients.This article reviews the incidence,diagnostic criteria,influence factors and prevention and treatment of early hematoma enlargement in intracerebral hemorrhage.

Objective:To evaluate the bone alteration subject to remodeling and analyze the esthetic result following immediate implant placement of incisors.Methods:In this study,20 patients (1 3 women,7 men)were involved,who needed implants for incisors of maxilla.The patients received 23 im-mediate implants totally.On the day of surgery and 6 months after the implants were placed,Cone beam CT (CBCT)was taken.The thickness of the alveolar ridge and the vertical change of marginal bone levels onthe mesial and distal aspects of theimplants were measured using the computer software (Planme-caRomexis Viewer 3.6.0.R).The evaluation of esthetic result by labial convexity score (LCS)and pa-pilla index score (PIS)were analyzed pre-operation and one year after the final crown was delivered.The statistics with paired-t test for the measurement data and Willcoxon test for rating data were done by SPSS 20.0.Results:The survival rate in the two-year follow-up was 1 00%.The measuring point 1 (MP1 ), MP2,MP3 and MP4 (0,2,4,6 mm apical to the implant platform,respectively)got significant altera-tions after 6 months of the follow-up.These differences were statistically significant (P<0.05 ).The major alteration happened at MP1 and MP4,which got (-0.89 ±2.06)mm and (-0.75 ±1 .28)mm reduction of the alveolar,respectively.The marginal alveolar ridge resorption was (-0.42 ±1 .24)mm and(-0.91 ±1 .96)mm for Ankylos System and Replace System,respectively,and the difference was not statistical significant .The esthetic results were quite acceptable.Before treatment,1 8 incisors rated 3 for LCS,and 2 incisors rated 4 for LCS;after final restoration,only 5 incisors rated 3 for LCS,and 1 4 incisors rated 2 for LCS.Before treatment,1 5 incisors rated 3 for PIS;after final restoration,1 3 incisors rated 3 for PIS.There was no statistically significant difference for the PIS pre-operation and 1 year after final restoration,while there was statistically significant negative change for LCS.Conclusion:Even fol-lowing the proper surgical technique,the alveolar ridge wall still can’t be maintained after immediate im-plant placed in fresh socket of incisors.The inter-dental papilla could be well maintained,while due to the remodeling of labial bone,labial convexity will inevitably collapse.Therefore immediate implant still has esthetic risk.

Objective:To summarize the clinical characteristics, diagnosis and treatment of caspase recruitment domain-containing protein 9 (CARD9) gene deficiency associated invasive candidiasis, and report a novel mutation in CARD9 gene.Methods:The clinical characteristics, laboratory tests, treatment and the outcome of follow-up in a boy with invasive candidiasis were described. The boy′s main clinical manifestations were central nervous system infection and retroperitoneal mass. Whole-exome sequencing was performed and Sanger sequencing was verified to identify the CARD9 gene mutations in the patient and his parents. A literature search for “CARD9”and “invasive candidiasis”was conducted in PubMed, Wanfang and CNKI databases from their establishment to May 2020.Results:A 10-year-old boy suffered onset symptom of chronic diarrhea, which lasted for two months. The symptom was followed by progressive neurological symptoms such as headache, vomiting, seizures and disorder of consciousness. His unusual medical history was absent. Candida albicans were cultured several times in cerebrospinal fluid and blood, and yeast-like fungi were found in the stool high power field of vision. Cerebral magnetic resonance imaging indicated obstructive hydrocephalus and abdominal CT scan showed retroperitoneal mass and thickening of the intestinal wall. The whole-exome sequencing analyses of blood samples from the boy and his parents were performed. The results showed that there was a homozygous mutation of c.952-12_956delinsAG in the CARD9 gene, which was an unreported pathogenic mutation. This was confirmed by Sanger sequencing. There was no significant relief from intravenous combined antifungal medications. After lateral ventricular drainage surgery and injection of amphotericin B into the lateral ventricle, improvement of clinical symptoms and cerebral spinal fluid abnormalities was observed after nine weeks, and the retroperitoneal mass shrank. At follow-up after four-month oral combined antifungal medications, the child had no complaint except fatigue. However, cerebral spinal fluid analysis showed increased protein level and decreased glucose. Persistent hydrocephalus and periventricular white matter abnormal signals were revealed on the brain magnetic resonance imaging and the smaller retroperitoneal mass than before on the abdominal CT scan. In addition to this case, totally 21 cases with CARD9 gene deficiency associated invasive candidiasis have been reported worldwide, most of which featured central nervous system infections.Conclusions:CARD9 gene deficiency is an autosomal recessive primary immunodeficiency that confers human susceptibility to fungal disease. The associated invasive candidiasis often affects the central nervous system and makes the patient severely ill. Adequate systemic antifungal therapies should be given, and patients with hydrocephalus need surgical treatment. A novel mutation is reported that expands the variant diversity of CARD9 gene. For patients with unexplained invasive candidiasis, including those without a history of previous recurrent infection, genetic testing is recommended for primary immunodeficiency including CARD9 gene deficiency.

AIM:To investigate the expression of Grb2-associated binding protein 2 (Gab2) in human osteo-sarcoma cells and its relationship with the invasion and metastases of human osteosarcoma cells.METHODS: The tech-nique of small RNA interference was used to transfect human osteosarcoma U2-OS cell lines.Western blotting and RT-PCR were used to detect the protein and mRNA expression of Gab2 in transfected U2-OS cells.After transfection, through chem-otaxis and invasion assays in vitro, the cell migration and invasion abilities were detected.RESULTS:After transfection, the expression of Gab2 at mRNA and protein levels in Gab2 siRNA transfected cells ( SiGab2/U2-OS) was lower than that in scrambled siRNA transfected cells ( Scr/U2-OS ) and U2-OS cells.After stimulation with epidermal growth factor ( EGF) at concentration of 10 μg/L, the migration SiGab2/U2-OS cells was significantly less than Scr/U2-OS cells and U2-OS cells ( P<0.01 ) .The number of invasion cells of SiGab2/U2-OS group was significantly lower than the other 2 control groups ( P<0.01) .CONCLUSION:Inhibition of Gab2 expression obviously attenuates the migration and invasion abilities of human osteosarcoma U2-OS cell line.

Objective To investigate the appropriate compatibility of appropriate compatibility of sevoflurane and propofol for patients with mild cognitive impairment (MCI) undergoing posterior lumbar interbody fusion in order to protect their cognitive function.Methods Eighty patients, 41 males, 39 females, aged 65-75 years, BMI 17-26 kg/m2, ASA physical status Ⅰ or Ⅱ, scheduled to undergo elective posterior lumbar interbody fusion, were to be scored according to Montreal cognitive assessment (MoCA), mini mental state examination (MMSE), dementia scale (CDR) and daily living ability scale (ADL) to identify patients with MCI before the surgery.They were randomly assigned to 4 groups (n=20 each) using a random number table: TCI propofol 2.0-2.5 μg/ml group (group P), TCI propofol 1.2 μg/ml+sevoflurane 0.6 MAC group (group PS1), TCI propofol 0.6 μg/ml+sevoflurane 0.9 MAC group (group PS2), 1.0-1.5 MAC sevoflurane group (group S).MoCA and MMSE were used to evaluate the cognitive function of patients 1 d before the operation (T0), after patients become wide-awake (T1), 3 d and 7 d after operation (T2 and T3).Apolipoprotein J (ApoJ) concentration related to cognitive function in blood samples, which were drawn at T0-T3 would be measured with ELISA method.Results Compared with T0, the scores of MMSE and MoCA in four groups decreased significantly (P＜0.05) at T1, the scores of MMSE and MoCA in group S decreased significantly (P＜0.05) at T2;compared with T1, the score of MMSE in the four groups increased significantly at T2, T3 (P＜0.05).The scores of MMSE at T1, T3 in group S decreased significantly compared with groups P, PS1 and PS2 (P＜0.05).The scores of MoCA at T2, T3 in group S decreased significantly compared with groups P, PS1 and PS2 (P＜0.05).Compared with T0, the concentration of plasma ApoJ in the four groups increased significantly at T1 (P＜0.05).Compared with T1, the concentration of plasma ApoJ in the four groups decreased significantly at T2 and T3 (P＜0.05).Compared with group PS1, the concentration of plasma ApoJ at T1, T3 increased significantly in groups S and group PS2 (P＜0.05).Conclusion TCI propofol 1.2 μg/ml combined with 0.6 MAC sevoflurane group is the appropriate compatibility of sevoflurane and propofol for patients with MCI undergoing posterior lumbar interbody fusion,because it has less negative influence on cognitive function and lower concentration of plasma ApoJ.

OBJECTIVETo compare clinical efficacy of Zero-profile implant for anterior cervical discectomy and fusion and conventional titanium plate with cage internal fixation for the treatment of single segmental cervical intervertebral disc herniation.

METHODSFrom August 2011 to March 2014, clinical data of 139 patients with single cervical disc herniation treated with anterior cervical discectomy and interbody fusion with internal fixation were retrospectively analyzed. The patients were divided into two groups according to its operation method. There were 63 patients in group A which performed anterior discectomy and interbody fusion with Zero-profile;76 patients in group B which performed anterior cervical discectomy and cage plate internal fixation. JOA score and Odom functional rating between two groups were compared before and after operation. Videofluorographic swallowing study (VFSS) were used to evaluate thickness of prevertebral soft tissue. Bazaz dysphagia score were used to assess incidence of dysphagia. Postoperative AP X-ray and CT of cervical vertebra at 12 months were applied for evaluating bone graft fusion. Postoperative MRI was applied for evaluating the incidence of adjacent segment degeneration. Blood loss,operative time, preoperative and postoperative JOA score, Odom functional rating and VFSS score, Bazaz score, fusion rate between vertebral bodies and incidence of adjacent segment degeneration were compared between two groups.

RESULTSThere were no statistical meaning between two groups in JOA score, Odom functional rating before and after operation (P > 0.05); and no significant meaning in VFSS score between two groups before operation (P > 0.05); There were no significant difference in operative time and blood loss. There was statistical meaning in VFSS, Bazaz dysphagia score at 2 days, and 6 months after operation (P < 0.05). All patients obtained bone union at 1 year after operation, and no obvious meaning in fusion rate (P > 0.05). Eight patients (12.7%) in group A occurred adjacent segment degeneration and 19 patients (25%) in group B occurred adjacent segment degeneration, and there was significant meaning between two groups (P < 0.05).

CONCLUSIONBoth of Zero-profile implant for anterior cervical discectomy and fusion and conventional cage internal fixation for the treatment of single segmental cervical intervertebral disc herniation could obtain satisfied clinical results. While Zero-profile implant for anterior cervical discectomy and fusion has advantages of lower incidence of adjacent segment degeneration, and its mid and long term following-up results still further observation.

Adult ; Aged ; Bone Plates ; Case-Control Studies ; Cervical Vertebrae ; surgery ; Diskectomy ; Female ; Fracture Fixation, Internal ; Humans ; Intervertebral Disc ; surgery ; Intervertebral Disc Displacement ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fusion ; Treatment Outcome