Objective To prepare rabbit polyclonal antibody against mouse IQ and ubiquitin-like domain-containing protein (IQUB) and detect its expression in the mouse testis. Methods Full-length coding sequence of IQUB was inserted into the pET-30a(+) vector to construct pET-30a-IQUB recombinant prokaryotic plasmid. Transformation of pET-30a-IQUB plasmid into E. coli BL21 was performed, and protein expression was induced with isopropyl-beta-D-thiogalactoside (IPTG). The protein was purified through histidine-tagged fusion protein purification column, then denatured by treatment of urea with gradient concentration. New Zealand rabbits were immunized with the denatured protein to produce IQUB polyclonal antibody. Antibody titer was detected by ELISA, and Western blot analysis and immunofluorescence assay were employed to validate the effectiveness and specificity of IQUB antibody. Results pET-30a-IQUB recombinant plasmid was constructed, and protein expression of IQUB was induced successfully with IPTG. The titer of IQUB polyclonal antibody reached 1:1 000 000. The antibody specifically recognized the endogenous IQUB protein of testis in the wild-type adult mouse. IQUB was expressed in spermatogenic cells of different stages. It was localized in the acrosome and flagellum of mature sperms. Conclusion The highly specific rabbit anti-mouse IQUB polyclonal antibody is successfully prepared, which can be used for Western blot and immunofluorescence histochemistry.
Male ; Rabbits ; Animals ; Mice ; Ubiquitins ; Escherichia coli/genetics* ; Isopropyl Thiogalactoside ; Antibodies ; Enzyme-Linked Immunosorbent Assay

Objective To produce rabbit polyclonal antibody against mouse testis expressed 38 (TEX38). Methods Full-length open reading frame sequence of TEX38 was amplified and inserted into the pET-30a-(+) vector to construct pET-30a-TEX38 prokaryotic plasmid. The recombinant plasmid was transformed into E.coli BL21, and expression was induced with isopropyl β-D-thiogalactopyranoside (IPTG). New Zealand white rabbits were immunized with TEX38 protein after purification and denaturation, then TEX38 polyclonal antibodies were collected from rabbit serum samples. ELISA was performed to detect the antibody titer. Western blot and immunofluorescence staining were performed to determine the specificity of TEX38 polyclonal antibodies. Results The pET-30a-TEX38 recombinant plasmid was constructed, and TEX38 prokaryotic protein was expressed and purified successfully. After immunization, the titer of TEX38 antibody reached 1:1 000 000. Western blot analysis and immunofluorescence staining showed that TEX38 was localized in the mouse spermatogenic cells and sperms with a good specificity. Conclusion The rabbit polyclonal antibody against mouse TEX38 is successfully produced, and the expression of TEX38 in mouse spermatogenic cells and sperms is validated.
Male ; Rabbits ; Animals ; Mice ; Testis ; Antibodies ; Enzyme-Linked Immunosorbent Assay ; Immunization ; Spermatozoa ; Escherichia coli

Objective:To compare the efficacy of series casting with different timings after botulinum toxin injection among equinus children with spastic cerebral palsy.Methods:Sixty-three equinus children with spastic cerebral palsy were randomly divided into a control group and an experimental group. After being injected with botulinum toxin type A, 4 weeks of series casting was applied immediately in the control group, while in the experimental group it was applied 2 weeks later. Both groups were given basic rehabilitation therapy including heel walking, kicking balls, single-leg standing, standing platform training and balance training. The modified Tardieu scale, three-dimensional gait analysis, gross motor function measure 88 (GMFM-88) and a self-made questionnaire were used to evaluate muscle tone, spasticity, gait parameters, standing and walking before the treatment and 2 and 6 months afterward.Results:Immediately after the treatment there was a significant improvement in both groups′ average MTS scores (R1, R2, R1-R2 with knee extension and flexion), dorsiflexion at initial contact, peak dorsiflexion in the stance and swing phases, and dimension D (standing), and also in their average GMFM-88 (walking) scores. Two and six months later the experimental group′s average MTS scores, peak dorsiflexion in the stance phase, and dimension D (standing) score in the GMFM-88 (walking) had improved significantly compared with the control group′s averages. However, no significant differences were observed between the two groups in any of the other measurements after the treatment.Conclusion:Delaying series casting after botulinum toxin A injection can significantly reduce tone and spasticity in the plantarflexors, improve the range of motion and functioning of the ankle, and alleviate the discomfort and pain caused by series casting.

Objective: To investigate the expression of SMARCA4 (BRG1) and SMARCB1 (INI-1) protein in endometrial dedifferentiated carcinoma (DDC) and undifferentiated carcinoma (UDC), and their correlation with clinicopathologic features. Methods: Clinicopathological information was gathered for 26 cases of DDC and UDC and consulting hospitals from January, 2006 to December, 2018 in Fudan University Shanghai Cancer Center, including 10 cases of DDC and 16 cases of UDC. Morphologic features and diagnosis were reviewed by two pathologists. Immunohistochemistry for expression of BRG1 and INI1 protein was performed. The correlations with clinicopathologic features were analyzed. Results: BRG1 and INI1 loss were present in 14 of 26 cases of DDC/UDC, including 12 BRG1-deficient cases and 2 INI1-deficient cases, respectively. Six cases demonstrated variable amounts of rhabdoid cells in 14 BRG1/INI1-deficient cases, and only 1 case showed rhabdoid cells in the 12 intact expression cases. However, there was no significantly statistical difference (P=0.060). Age, invasive depth, lymph node status and FIGO stage were not associated with the expression of the BRG1 and INI1 (P=0.437, P=0.672, P=0.242, P=0.348). Remarkably, the BGR1/INI1-deficient patients had worse survival than those with intact expression (4.7 vs. 22.9, P=0.033). Conclusion BRG1/INI1-deficient is observed in approximately half of DDC and UDC. Identification of these tumors is clinically relevant due to their more aggressive behavior and poor prognosis. Hence, BRG1 and INI1 immunohistochemical stains should be performed for DDC and UDC in order to help the pathologists to distinguish these tumors from other carcinomas, and to predict the clinical prognosis.

Delirium is a common complication during perioperative period of lung transplantation, which affects postoperative recovery and survival. This article reviews the general situation of delirium during perioperative period of lung transplantation and the risk factors of perioperative delirium associated with lung transplantation, and proposes comprehensive nursing intervention measures to prevent delirium, in order to provide a reference for the nursing of perioperative delirium in lung transplantation patients.

Objective To investigate the expression of SMARCA4 (BRG1) and SMARCB1 (INI?1) protein in endometrial dedifferentiated carcinoma (DDC) and undifferentiated carcinoma (UDC), and their correlation with clinicopathologic features. Methods Clinicopathological information was gathered for 26 cases of DDC and UDC and consulting hospitals from January, 2006 to December, 2018 in Fudan University Shanghai Cancer Center, including 10 cases of DDC and 16 cases of UDC. Morphologic features and diagnosis were reviewed by two pathologists. Immunohistochemistry for expression of BRG1 and INI1 protein was performed. The correlations with clinicopathologic features were analyzed. Results BRG1 and INI1 loss were present in 14 of 26 cases of DDC/UDC, including 12 BRG1?deficient cases and 2 INI1?deficient cases, respectively. Six cases demonstrated variable amounts of rhabdoid cells in 14 BRG1/INI1?deficient cases, and only 1 case showed rhabdoid cells in the 12 intact expression cases. However, there was no significantly statistical difference (P=0.060). Age, invasive depth, lymph node status and FIGO stage were not associated with the expression of the BRG1 and INI1 (P=0.437,P=0.672,P=0.242,P=0.348). Remarkably, the BGR1/INI1?deficient patients had worse survival than those with intact expression (4.7 vs. 22.9, P=0.033). Conclusion BRG1/INI1?deficient is observed in approximately half of DDC and UDC. Identification of these tumors is clinically relevant due to their more aggressive behavior and poor prognosis. Hence, BRG1 and INI1 immunohistochemical stains should be performed for DDC and UDC in order to help the pathologists to distinguish these tumors from other carcinomas, and to predict the clinical prognosis.

BACKGROUND AND PURPOSE: External counterpulsation (ECP) is a noninvasive method used to enhance cerebral perfusion by elevating the blood pressure in ischemic stroke. However, the response of the beat-to-beat blood pressure variability (BPV) in ischemic stroke patients during ECP remains unknown. METHODS: We enrolled recent ischemic stroke patients and healthy controls. Changes in the blood flow velocities in bilateral middle cerebral arteries and the continuous beat-to-beat blood pressure before, during, and after ECP were monitored. Power spectral analysis revealed that the BPV included oscillations at very low frequency (VLF; <0.04 Hz), low frequency (LF; 0.04-0.15 Hz), and high frequency (HF; 0.15-0.40 Hz), and the total power spectral density (TP; <0.40 Hz) and LF/HF ratio were calculated. RESULTS: We found that ECP significantly increased the systolic and diastolic blood pressures in both stroke patients and controls. ECP decreased markedly the systolic and diastolic BPVs at VLF and LF and the TP, and the diastolic BPV at HF when compared with baseline. The decreases in diastolic and systolic BPV reached 37.56% and 23.20%, respectively, at VLF, 21.15% and 12.19% at LF, 8.76% and 16.59% at HF, and 31.92% and 23.62% for the total TP in stroke patients, which did not differ from those in healthy controls. The change in flow velocity on the contralateral side was positively correlated with the total TP systolic BPV change induced by ECP (r=0.312, p=0.035). CONCLUSIONS: ECP reduces the beat-to-beat BPV when increasing the blood pressure and cerebral blood flow velocity in ischemic stroke patients. ECP might be able to improve the clinical outcome by decreasing the beat-to-beat BPV in stroke patients, and this should be explored further in future studies.
Blood Flow Velocity ; Blood Pressure* ; Cerebrovascular Circulation* ; Counterpulsation* ; Humans ; Methods ; Middle Cerebral Artery ; Perfusion ; Stroke*

Objective To investigate the possible formation mechanism and imaging features of the hyperintense vessel sign (HVS) on fluid attenuated inversion recovery (FLAIR) in patients with ischemic stroke or transient ischemic attack (TIA).Methods The baseline data of the patients with middle cerebral artery (MCA) ischemic stroke or TIA with digital subtraction angiography (DSA) showing the lesions of MCA M1 segment in clinical practice were retrospectively retrieved from Nanjing Stroke Registry Program from January 2010 to July 2011.FLAIR was used to observe HVS,and DSA was used to evaluate the degree of vascular stenosis and cerebral collateral circulation.Results A total of 101 patients were enrolled,76 (75.2％) were males,and their mean age was 53.94 ± 13.47 years; 90 patients (89.1％) with ischemic stroke and 11 patients (10.9％) with TIA; 55 patients (54.5％) were HVS negative and 46 (45.5％) were HVS positive.Among the patients whose MCA stenosis ＜50％,50％-70％,70％-90％ and ≥90％,the positive rates were 0％ (0/8),25.0％ (3/12),17.6％ (3/17),and 62.5％ (40/64),respectively.There were significant differences (Z=-4.479,P＜ 0.001).The leptomeningeal collateral circulation of the HVS positive group was significantly more than that of the HVS negative group (Z =-6.196,P ＜ 0.001).Multivariate logistic regression analysis showed that the degree of MCA stenosis was an independent risk factor for influencing the formation of HVS (odds ratio 3.943,95％ confidence interval 2.03-7.659; P ＜0.001).Conclusions The formed intracranial leptomeningeal colhteral circulation after severe intracranial vascular stenosis or occlusion is a major pathophysiological basis of HVS formation on FLAIR sequences in patients with ischemic stroke or TIA.

Objective To investigate the significance of the Children's Rehabilitation Management System (ICR 2.0) for occupational therapy (OT) for cerebral palsy (CP). Methods 60 children with CP were divided into group A (n=30) and group B (n=30). The group A accepted OT program assisted with ICR 2.0, and the group B accepted OT program drawn by therapists experientially. They were assessed with Enjoji infant development scale and Fine Motor Function Measure (FMFM) before and 3 months after treatment. Results There was no significant difference in development months and the fine motor between 2 groups before treatment (P>0.05). Both the development and fine motor improved after treatment (P<0.001), and improved more in the group A than in the group B (P<0.05). Conclusion ICR 2.0 assisted OT program can further improve the recovery of children with CP.

Objective To reveal the incidence and types of acid-base disorder (ABD) of critically ill patients in emergency.Methods The clinical data of critically ill patients in the resuscitation room were collected prospectively from December 1,2008 to March 31,2009.Both arterial and venous blood samples were collected to detect arterial blood gas analysis and serum electrolytes simultaneously and their ABD were analyzed.Results Of totally 766 cases,the incidence of ABD was 97.3％ (745 cases).Among ABD the simple acid-base disorder ( SA BD) was 149 cases ( 20.0％ ),dual acid-base disorder (DABD) 525 (70.5％ ) and triple acid-base disorder (TABD) 71 (9.5％ ).After calculating anion gap (AG),the incidence of TABD increased from 12 to 71 cases and the missed diagnosis rate were 83.1％ (59 cases) and after calculating potential HCO3-,the incidence of TABD increased from 8 to 71 cases and the missed diagnosis rate were 88.7％ (63 cases).The age of patients with TABD (77.9 ± 10.7 years old) was older ( P ＜ 0.01 ) than that of other two groups of SABD and DABD as well as APACHEⅡ score (25.9 ± 7.1 ) and incidence (22.5％ ) of MODS in TABD were higher (P ＜0.01 ).The mortality of patients with TABD on the first,second,third and seven day were 14.1％,23.9％,26.8％ and 38.0％ respectively and significantly higher than those in SABD and DABD (P ＜0.05 or P ＜0.01 ).Conclusions The incidence of ABD was 97.3％.Calculation of AG and potential HCO3- would help find metabolic acidosis and TABD.The age and APACHEⅡ score were key factors to poor prognosis of patients with TABD.