Objective A high performance liquid chromatographic (HPLC) method was established for the determination of glycerol in propofol medium and long chain fat emulsion injection.MethodsThe chromatographic conditions were as follows: Kromasil 100-5-NH2 column(4.6×250mm,5μm) with the column temperature was 40℃,acetonitrile-water(8515)as mobile phase with flow rate of 1.0mL/min.Glycerol was detected by refractive index (RI) detector at 40℃.ResultsThe linear range of glycerol was 455.3916-2276.9580μg/mL(r=0.9999,n=7),the average recovery rate was 99.5%,RSD was 0.6%(n=9),the limit of detection(LOD) was 121ng and the limit of quantification(LOQ)was 364ng.ConclusionThe method was simple, rapid, strong specifity and accurate with good reproducibility, which is suitable for the content determination of glycerol in propofol medium and long chain fat emulsion injection.

Objective To study the effect of de novo donor-specific antigen (DSA) on transplant heart function and recipient survival after heart transplantation.Methods 195 recipients went through heart transplantion from March 2013 to January 2016 in our center,13 patients with preoperative panel reactive antibody (PRA) and 10 patients suffered from in-hospital death were exclude from this study,and the actual number of patients enrolled in this study was 172.By detecting HLA typing and DSA,recipients were divided into DSA positive group,anti-HLA antigen positive but DSA negative group (DSA negative group) and anti-HLA negative group.Cardiac dysfunction,coronary heart disease and cardiovascular death were recorded as cardiac events during the follow-up period.By analyzing the cardiac events rate among 3 groups,the relationship between DSA and cardiac events was acknowledged.Results The mean follow-up period of all patients was (1.3 ± 0.8) years.Among 172 patients,6 were positive for DSA (3.4％).In the DSA positive group,66％ of DSA were directed at HLA Ⅱ,mainly against HLA-DQ,1 developed only anti-HLA I antibody,1 developed both anti-HLA Ⅰ and Ⅱ antibody.The median developing time of DSA was (256 ± 125) days,and the distribution was centralized in the first half year.84％ of patients in DSA positive group were witnessed cardiac events.One patient was diagnosed as coronary heart disease and received PCI at 145th days after DSA was developed.Four out of 6 patients experienced cardiac dysfunction with median developing time of (54 ± 13) days,and the cardiac function restored after treatment with immunosuppression modification,high-dose glucocorticoid and IVIG.In the DSA negative PRA positive group,the incidence of cardiac events was 13％.There was one cardiovascular death,and 2 cases of cardiac dysfunction.In the HLA antigen negative group,the incidence of cardiac events was 4％.Cox regression analysis revealed that DSA could be seen as an independent risk factor in leading to cardiac events and affecting mid-long term survival of recipients (P =0.02).Significant difference was observed in Kaplan-Meier analysis among 3 groups (P＜0.001).Conclusion DSA has strong impact on outcome after heart transplantation.Routine surveillance and appropriate treatment should be attached to DSA.

Objective To observe the clinical effect of monosialotetrahexosyl ganglioside sodium injection (GM1) on spastic cerebral palsy. Methods 98 children with spastic cerebral palsy were randomly divided into control group (n=50) and treatment group (n=48). Both groups received Bobath approach, and the treatment group received GM1 in addition. They were assessed with Functional Independence Measure for Children (WeeFIM), Gross Motor Function Measure (GMFM) and Gesell Development Schedule (GDS) before and after 90 days of treatment. Results The scores of WeeFIM, all the dimensions of GMFM and the gross motor, fine motor, personal-social and adaption of the GDS improved in both groups after treatment (P<0.05), and improved more in the treatment group than in the control group (P< 0.05). Conclusion GM1 may further improve the recovery of function for children with spastic cerebral palsy.

Objective:To explore the angiographic distribution of the anomalous ostium of right coronary artery (RCA) and improve the technical tips for catheter incubation during right coronary angiography.Methods:A total of 24 861 patients who underwent coronary angiography in Zhongshan Hospital,Fudan University from May 2015 to May 2017 were reviewed,and the cases with anomalous RCA origin were identified.The aortic root at left anterior oblique 40°-45° view during coronary angiography was set as an imaginary plane,which was divided into four quadrants.The ostia distributions of anomalous RCAs,the image features and the technical essentials were analyzed.Results:Totally 147 cases had anomalous RCA origin and all underwent coronary angiography successfully.Among them,67 patients (45.6％,Type A) had an ectopic RCA origin from left sinus of valsalva (LSOV),48 patients (32.7％,Type B) had an anomalous RCA origin from left side of ascending aorta,and 32 patients (21.7％,Type C) had an anomalous RCA origin from right side of ascending aorta.In angiography image,some special signs named as "sea gull sign" and "horn sign" were observed in cases with anomalous RCA origin from LSOV or aortic sinus crest.Such as transposition of the heart and widened aortic root,the anomalous RCA originated from ascending aorta near coronary sinus in angiography image were actually originated from LSOV in some cases.Conclusions:The anomalous RCA arising from LSOV is the most common variation,followed by arising from ascending aorta.The RCA arising from ascending aorta above right coronary sinus is relatively rare.Understanding of the distribution regularity and image features of ectopic RCAs is helpful for coronary angiography and intervention.

OBJECTIVETo observe the effects of cryopreserved (-70 degrees C) neural stem cells (NSCs) transplantation on the axon regeneration after the spinal cord injury (SCI) of rats.

METHODSNeural stem cells were cultured from the hippocampus of the rat's embryo and identified by immunocytochemistry of nestin, then NSCs in logarithmic phage were cryopreserved (-70 degrees C) for 2 weeks. Seven days after the operation of SCI, the NSCs in were transplanted into the injured site immediately. Thirty six adult Wistar rats were randomly divided into three groups: spinal cord injury treated with transplantation of NSCs to the injured site (Group A), spinal cord injury received DMEM solution (Group B), control group (Group C). Then NSCs labeled with Brdu were detected by immunohistochemisty, and the reconstruction of spinal cord were detected by horseradish peroxidase (HRP) staining.

RESULTSNSCs can be detected in the spinal cord after transplantation. The number of HRP positive cells of Group A was higher than that of Group B.

CONCLUSIONThe transplantation of cryopreserved NSCs can survived in the injured site and promote the reconstruction. It may be a progress in the repairing of the SCI by cells transplantation.

Animals ; Axons ; physiology ; Cryopreservation ; Male ; Nerve Regeneration ; Rats ; Rats, Wistar ; Spinal Cord Injuries ; physiopathology ; therapy ; Stem Cell Transplantation

OBJECTIVETo investigate the effect of polycaprolactone-ascobic acid (PCL-AA) scaffolds in promoting repair of articular cartilage defects in a rabbit model.

METHODSThe cartilage defects (3.5 mm in diameter and 3.0 mm in depth) were created in the trochlear groove of the bilateral knees of eight 6-month-old male New Zealand white rabbits. The rabbit models were then randomized into 3 groups to receive implantation of PCL-AA scaffolds (group A, n=8), implantation of PCL scaffolds without AA (group B, n=5), or no treatment (group C, n=3). In groups A and B, the mixture of fibrin gel (10 µg) and thrombinogen (10 µg) was injected into the defects to fix the scaffolds during the surgery. Histological analyses and quantitative assessments of defect repair were conducted at 6 and 12 weeks after implantation of the scaffold.

RESULTSAt 6 weeks after scaffold implantation, macroscopic observation showed better filling of the cartilage defects in group A than in group B, while no obvious defect repair was observed in group C. The rabbits in group A showed a significant improvement of the Wakitani score than those in group B (4.05∓1.11 vs 7.05∓0.98, P<0.05). HE staining revealed the presence of newly generated cells in and around the PCL-AA scaffolds without inflammatory cells. Safranin O staining showed a significantly greater ECM of the newly regenerated tissue in groups A and B than in group C (P<0.05), and the volume of the regenerated cartilage and cells was significantly greater in group A than in group B (P<0.05). Samples harvested at 12 weeks showed more hyalione-like cartilage formation than that at 6 weeks in group A.

CONCLUSIONPCL-AA scaffolds have a good biocompatibility and promotes the healing of articular cartilage defects. Adding ascorbic acid into PCL scaffolds better promotes cartilage formation in terms of both quantity and quality of the regenerated tissues. PCL-AA scaffolds can serve as a promising biomaterial to promote the regeneration of articular cartilage using tissue engineering techniques.

Diarrhea is a common intestinal symptom in macaque.The corresponding intestinal lesions of macaque are mainly described at autopsy but less observed by colonoscopy.The aim of this study was to develop a colonoscopic technique and to obtain endoscopic images of the entire colon in macaques.Eight healthy adult macaques ( 5 males and 3 females) without diarrhea for 2 months, were fed Glauber’ s salt through nasogastric tubes.The colon cleanliness was well matched to the endoscopic observation of macaque colon.The procedure took 10-20 min for each animal.There was no obvious abnormality in the colon of four animals except some slight differences of mucosal structure from that of human beings.Small pieces of erosion and ulcer in the colons were observed in four macaques which presented mild diarrhea for less than 1 day, while a severe stenosis was observed in one of those four macaques.No animal died during and one week after the endoscopic procedure.Colonoscopy may safely performed in macaques.The images taken by colonoscopy may be important to establish diagnosis and treatment of colitis in macaques in time and to evaluate the efficacy of drug intervention as well.This technique is also helpful to provide qualified macaques for scientific researches.

To investigate the freshness, high molecular weight substances, the determination of polypeptide, haemolysis and agglomeration, biological activity of Cervus and Cucumis polypeptide injection; to provide the direction for improving the quality of products for enterprises; furthermore, to provide reference for the revision of the quality standards of Cervus and Cucumis polypeptide injection. Firstly, we investigated the factors affecting the freshness of the injection, including biogenic amines, aflatoxins, the acid value and peroxide value of the melon seeds. The method of dansyl chloride pre-column derivatization-HPLC was used to determine the content of 8 biogenic amines in Cervus and Cucumis polypeptide injection. The method validation results showed good specificity, precision, linearity and recovery rates, which was suitable for the determination of biogenic amines in Cervus and Cucumis polypeptide injection. The results of sample determination showed that relatively higher concentrations of cadaverine were detected in the products from company B. The results of aflatoxins, acid value and peroxide value showed that the melon seeds from some companies had rancidity, mildew and other problems, indicating that the quality standards of multi-component biochemical drugs containing animal- and plant-derived components should be controlled in terms of freshness. Secondly, the methods for the determination of high molecular weight substances and polypeptides in the quality standard were improved. Tricine-SDS-PAGE electrophoresis was used instead of gel chromatography to determine the high molecular weight substances, which improved the accuracy of determination. The kits were used instead of folin-phenol for the determination of peptide content, which is easy to operate, specific and suitable for high-throughput sample determination. Finally, the haemolysis, agglomeration, and biological activity of Cervus and Cucumis polypeptide injection were studied. The results showed that no haemolysis and agglomeration were found in all samples, and the inhibitory effect of samples on THP-1 proliferation in vitro from different companies was different to some extent. In conclusion, the optimized quality standard is more suitable for the detection of Cervus and Cucumis polypeptide injection, and can lay the foundation for improving the safety of multi-component biochemical drugs.

OBJECTIVE: To explore the coincidence rate of G-banding karyotype analysis and fluorescence in situ hybridization (FISH) for the diagnosis of children with sex chromosome mosaicisms. METHODS: A retrospective analysis was carried out for 157 children with suspected sex chromosome abnormalities who had presented at Shenzhen Children's Hospital from April 2021 to May 2022. Interphase sex chromosome FISH and G-banding karyotyping results were collected. The coincidence rate of the two methods in children with sex chromosome mosaicisms was compared. RESULTS: The detection rates of G-banding karyotype analysis and FISH were 26.1% (41/157) and 22.9% (36/157) , respectively (P > 0.05). The results of G-banding karyotype analysis showed that 141 cases (89.8%) were in the sex chromosome homogeneity group, of which only 5 cases (3.5%) were inconsistent with the results of FISH. There were 16 cases (10.2%) in the sex chromosome mosaicism group, of which 11 cases (68.8%) were inconsistent with the results of FISH. There was a statistical difference between the two groups in the coincidence rate of the results of the two methods (P < 0.05). CONCLUSION No significant difference was found between G-banding karyotype analysis and FISH in the detection rate of chromosome abnormalities. The coincidence rate in the mosaicism group was lower than that in the homogeneity group, and the difference was statistically significant. The two methods should be combined for clinical diagnosis.
Humans ; Mosaicism ; In Situ Hybridization, Fluorescence/methods* ; Retrospective Studies ; Karyotyping ; Chromosome Aberrations ; Sex Chromosome Aberrations ; Karyotype ; Chromosome Banding ; Sex Chromosomes

Stem cells play an important role in self-renewal and repair of tissues and organs.Under the influence of the external magnetic field, labeling the stem cells with magnetic nanoparticles can increase the directional homing and retention of the stem cells, thereby amplifying the repair effect of the stem cells.In this paper, the principle of magnetic nanoparticles targeting stem cells and its progress in the application of various pathological models are reviewed.