The purpose of this study was to synthesize and evaluate the necrosis target of MRI contrast agent based on rhein.The novel ligand 10-{ [6-(1,8-dihydroxyanthraquinone-3-carboxamido) hexyl] amino} acetyl-1,4,7,10-tetraazacyclododecan-1,4,7-triacetic acid (DO3A-rhein) was synthesized by two-step acylation and two-step deprotection.The paramagnetic contrast agent gadolinium 10-{ [6-(1,8-dihydroxyanthraquinone-3-carboxamido) hexyl] amino} acetyl-1,4,7,10-tetraazacyclododecan-1,4,7-triacetate (Gd-DO3A-rhein) was obtained by coordination of Gd3+ with the synthesized ligand.Its necrosis affinity was evaluated by liver infarction and muscular necrosis on rat models.The MRI was performed before administration of Gd-DO3A-rhein and during 0 h to 12 h after administration of Gd-DO3A-rhein (0.1 mmol/kg),respectively,and Gd-DOTA was used as control.After MRI scanning,rats were sacrificed and necrotic tissues were stained using triphenyltetrazolium chloride (TTC) and hematoxylin-eosin (HE).MRI images of liver infarction and muscular necrosis on rat models showed significantly enhanced signal intensity compared with normal tissues.The contrast ratios of necrotic liver/normal liver were 1.61 ±0.14 and 2.36 ±0.20 at 3 h and 12 h postinjection of Gd-DO3A-rhein (0.1 mmol/kg) respectively,demonstrating a significant difference compared with pre-administration of Gd-DO3A-rhein (1.16 ±0.10;P ＜ 0.05).The same results were obtained from necrotic muscles.These findings suggested that Gd-DO3A-rhein possessed the necrosis target and imaging capability of necrotic tissues.

Objective To establish the rat model of collagen-induced arthritis and study the diagnostic value of MRI for early RA by comparing to pathological changes. Methods Thirty Wistar rats were randomly divided into normal groups and experimental groups.Fifteen rats of the experimental groups were immunized with type Ⅱ collagen and Freund's Adjuvant Incomplete. At the scheduled days, the selected rats of both experimental and normal groups underwent X-ray, 1.5T MR scan plus enhancement and histological analysis. Results Arthritis Index increased on the 14th day after immunization and reached the peak on the 28th day in the experimental group, the positivity of Anti-CⅡ antibody was significantly different from the normal group.The experiemental model was established in 93.33% of all rats. The enhanced MRI showed that 12 (12/15) rats presented with abnormal signs . The sensitivity of MRI was 85.71% and the specificity was 100%. There was no significant correlation could be found between MRI and histological changes X-ray revealed soft tissue swollen in 4 (4/15) rats, which showed that MRI had higher sensitivity in detecting abnormal signs of arthritis than X-ray. Conclusion MRI may be used in the early diagnosis of early RA.

Objective To establish a method to simultaneously determine syringing and isofraxidin by HPLC;To investigate the features of percutaneous absorption in vitro of Jiegugao blended and pasted by white vinegar, honey and vaseline; To discuss the mechanism of commonly used ointment matrices in Tujia Minority. Methods Rat abdomen skin in vitro was as transdermal barrier;the modified Franz diffusion pool was used to simulate human skin medication; the content of syringin and isoprofen was determined by HPLC; the percutaneous absorption equation was established and the related parameters, such as cumulative permeation rate and permeation rate, were calculated. Results When using Syncronis C18 (250 mm × 4.6 mm, 5 μm) as chromatographic column, acetonitrile-0.1%phosphoric acid as mobile phase, 1.0 mL/min as perfusion speed and 265 nm as determine wavelength, regression equation of syringingwas A=10 686.454 6C+1565.778 8 (r=1.000 0), regression equation of isofraxidin was A=12 297.305 4C-5913.729 9 (r=0.999 9). Cumulative permeation quantity of syringing in Jiegugao blended and pasted by white vinegar, honey, vaseline and blank were 7.549 2, 4.580 3, 3.890 8 and 5.378 4 μg?cm-2?h-1 respectively and permeation rate were 25.66%, 16.11%, 13.73% and 18.78%. Meanwhile, cumulative permeation quantity of isofraxidin were 2.536 9, 1.941 8, 1.178 2 and 2.293 6 μg?cm-2?h-1 respectively and permeation rate were 47.04%, 35.06%, 22.11%and 41.11%. Conclusion Using white vinegar as the ointment matrix can promote the percutaneous absorption of effective composition in Jiegugao blended. However, it will retard the percutaneous absorption of effective composition in Jiegugao when using honey and vaseline as the ointment matrices, but honey and vaseline can be used as a slow-release matrix.

BACKGROUND: In the treatment of osteoporotic vertebral compression fractures with percutaneous vertebroplasty, the efficacy and safety of bone cement injection by unipedicular and bipedicular approaches are still controversial. Some studies suggest that bone cement injection via unipedicular approach can shorten operation time and reduce postoperative complications, while the other studies suggest that bone cement injection via bipedicular approach can make bone cement distribute more evenly in the vertebral body and relieve pain better. OBJECTIVE: To systematically assess the efficacy and safety of percutaneous vertebroplasty via unipedicular versus bipedicular approach in the treatment of osteoporotic vertebral compression fractures. METHODS: Randomized controlled trials about unipedicular versus bipedicular percutaneous vertebroplasty in the treatment of osteoporotic vertebral compression fracture published before September 18 t h, 2018 were retrieved in the PubMed, Cochrane library, Embase, CNKI, VIP, WanFang data and CBM. Two researchers independently screened all the literatures, carried out data extraction and used improved Jadad to evaluate the methodological quality of the included studies. Meta-analysis using Revam 5.3 was conducted. Egger's test was utilized to evaluate the publication bias. RESULTS AND CONCLUSION: Totally 14 randomized controlled trials including 900 cases were eventually included, 452 cases in unipedicular approach group and 448 cases in bipedicular approach group. The Meta-analysis results showed that compared with the bipedicular approach, the unipedicular approach required shorter operation time [weighted mean difference (WMD) =-16.59, 95％ confidence interval (CI) (-19.25, -13.94), P < 0.001], smaller amount of bone cement injected [WMD=-1.27, 95％ CI (-1.64, -0.89), P < 0.001], and had lower incidence of cement leakage [relative risk=0.70, 95％ CI (0.53, 0.92), P =0.01]. There were no significant differences in short-and long-term Visual Analogue Scale scores, short-and long-term Oswestry Disability Index scores, and the postoperative incidence of adjacent vertebral fractures between the two groups (P> 0.05). Overall, bone cement injection both via bipedicular and unipedicular approaches can lead to a significant improvement in pain relief and living quality of osteoporotic vertebral compression fracture patients, but bone cement injection via unipedicular approach can shorten operation time, reduce cement volume and lower the incidence of cement leakage compared with the bipedicular approach.

Objective A finite element analysis was conducted on the biomechanics of the locking plate and intramedullary nail fixation for the treatment of distal tibial fractures,and the resuhs were verified combined with clinical cases,so as to provide references for clinical treatment.Methods (1) Finite element analysis:the three-dimensional CT data of the lower limbs of a healthy male volunteer were used to establish a finite element model.The internal stress distribution of the tibial plateau was set to 60％ of the total load by intramedullary nail and locking plate respectively,and the tibia end was fixed effectively.400 N axial pressure load which equaled to that of adult knee joint during single axis standing was simulated.The equivalent stress and displacement of the model by different fixations were compared.(2) Clinical verification:a retrospective case control study was performed on the clinical data of 37 cases of distal tibia1 fractures treated with internal fixation from June 2015 to December 2016,including 17 cases in intramedullary nail group and 20 in locking plate group.The operation time,intraoperative blood loss,postoperative fracture healing time,and postoperative Johner-Wruhs score of patients were recorded for comprehensive assessment of recovery.Results (1) The finite element analysis results:the maximum stress value was 5.907 MPa for intramedullary nail and 5.821 MPa for locking plate model (P ＞0.05),respectively.The maximum displacement of intramedullary nail model was 2.313 mm,lower than that of locking plate fixation system (3.854 rmm) (P ＜ 0.05).(2) Clinical verification:the operation time and intraoperative blood loss of intramedullary nail were both lower than those of locking plate [(114.1 ±21.6)minutes):(129.8±21.4)minutes and (152.9 ±64.88)ml:(212.5 ±98.5)ml](P ＜0.05).The average fracture healing time was (17.7 ± 2.8)weeks for intramedullary nail and (20.6 ± 4.1) weeks for locking plate (P ＜ 0.05),respectively.In the intramedullary nail group,the Johner Wruhs score was excellent in 13 cases and good in four cases,with excellent and good rate of 100％,while in the locking plate group,nine cases were excellent,eight were good,and three were fair,with excellent and good rate of 85％ (P ＞ 0.05).Conclusions In terms of biomechanics and clinical effect,intramedullary nail fixation is superior than the medial locking plate fixation for the treatment of the distal tibial fractures.Intramedullary nail fixation can reduce surgical trauma and bone displacement after fixation and promote fracture healing.

Objective: To analyze variations of TYR and P genes among 14 patients with clinically diagnosed oculocutaneous albinism. Methods: Potential variations of the TYR and P genes were detected by Sanger sequencing. Novel variations were predicted with bioinformatics software including SIFT and PolyPhen-2. Results: No variation was found in the TYR gene, while 9 types of variations were found in the P gene among the 14 patients, which included c. 803-3C>G (7/26), c. 1327G>A (p.Val443Ile) (5/26), c. 632C>T (p.Pro211Leu) (4/26), c. 1832T>C (p.Leu611Pro) (3/26), c. 1349C>A (p.Thr450Lys) (2/26), c. 2363C>T (p.Ser788Leu) (2/26), c. 2228C>T (p.Pro743Leu) (1/26), c. 1525A>G(p.Thr509Ala) (1/26), and c. 1349C>T(p.Thr450Met) (1/26). Only 1 heterozygous variation was detected in 2 families. c. 2363C>T (p.Ser788Leu), c. 1832T>C (p.Leu611Pro) and c. 1525A>G (p.Thr509Ala) were not reported previously and predicted as "harmful" to the protein function. Conclusion The main type of ocular albinism is oculocutaneous albinism type Ⅱ in Liuzhou region, where the most common variations of the P gene were c. 803-3C>G and c. 1327G>A (p.Val443Ile). Above finding has enriched the variation spectrum of the P gene.

OBJECTIVE: To carry out mutation analysis and prenatal diagnosis for a family affected with primary carnitine deficiency. METHODS: Genomic DNA of the proband was extracted from peripheral blood sample 10 days after birth. The 10 exons and intron/exon boundaries of the SLC22A5 gene were subjected to PCR amplification and Sanger sequencing. The proband's mother was pregnant again two years after his birth. Fetal DNA was extracted from amniocytes and subjected to PCR and Sanger sequencing. RESULTS: Tandem mass spectrometric analysis of the proband revealed low level of plasma-free carnitine whilst organic acids in urine was normal. Compound heterozygous SLC22A5 mutations c.1195C>T (inherited from his father) and c.517delC (inherited from his mother) were detected in the proband. Prenatal diagnosis has detected no mutation in the fetus. The plasma-free carnitine was normal after birth. CONCLUSION Appropriate genetic testing and prenatal diagnosis can prevent further child with carnitine deficiency. The identification of c.517delC, a novel mutation, enriched the spectrum of SLC22A5 mutations.
Cardiomyopathies ; genetics ; Carnitine ; deficiency ; genetics ; Child, Preschool ; DNA Mutational Analysis ; Female ; Humans ; Hyperammonemia ; genetics ; Muscular Diseases ; genetics ; Mutation ; Pregnancy ; Prenatal Diagnosis ; Solute Carrier Family 22 Member 5 ; genetics

OBJECTIVE: To detect variants of IVD gene among 4 neonates with suspected isovalerate acidemia in order to provide a guidance for clinical treatment. METHODS: 111 986 newborns and 7461 hospitalized children with suspected metabolic disorders were screened for acyl carnitine by tandem mass spectrometry. Those showing a significant increase in serum isovaleryl carnitine (C5) were analyzed for urinary organic acid and variants of the IVD gene. RESULTS: Four cases of isovalerate acidemia were detected, which included 2 asymptomatic newborns (0.018‰, 2/111 986) and 2 children suspected for metabolic genetic diseases (0.268‰, 2/7461). The formers had no obvious clinical symptoms. Analysis of acyl carnitine has suggested a significant increase in C5, and urinary organic acid analysis has shown an increase in isovaleryl glycine and 3-hydroxyisovalerate. Laboratory tests of the two hospitalized children revealed high blood ammonia, hyperglycemia, decreased red blood cells, white blood cells, platelets and metabolic acidosis. The main clinical manifestations have included sweaty foot-like odor, feeding difficulty, confusion, drowsiness, and coma. Eight variants (5 types) were detected, which included c.158G>A (p.Arg53His), c.214G>A (p.Asp72Asn), c.548C>T (p.Ala183Val), c.757A>G (p.Thr253Ala) and 1208A>G (p.Tyr403Cys). Among these, c.548C>T and c.757A>G were unreported previously. None of the variants was detected by next generation sequencing of 2095 healthy newborns, and all variants were predicted to be likely pathogenic based on the guidelines from the American College of Medical Genetics and Genomics. CONCLUSION The incidence of isovalerate acidemia in Liuzhou area is quite high. Screening of metabolic genetic diseases is therefore recommended for newborns with abnormal metabolism. The discovery of novel variants has enriched the mutational spectrum of the IVD gene.
Infant, Newborn ; Child ; Humans ; Acidosis ; Carnitine ; Erythrocytes ; High-Throughput Nucleotide Sequencing

Necrosis is a form of cell death, which is related to various serious diseases such as cardiovascular disease, cancer, and neurodegeneration. Necrosis-avid agents (NAAs) selectively accumulated in the necrotic tissues can be used for imaging and/or therapy of related diseases. The aim of this study was to preliminarily investigate necrosis avidity of I-evans blue (I-EB) and its mechanism. The biodistribution of I-EB at 24 h after intravenous administration showed that the radioactivity ratio of necrotic to viable tissue was 3.41 in the liver and 11.82 in the muscle as determined by counting in model rats. Autoradiography and histological staining displayed preferential uptake of I-EB in necrotic tissues. nuclear extracts from necrotic cells exhibited 82.3% of the uptake in nuclei at 15 min, as well as 79.2% of the uptake at 2 h after I-EB incubation. The DNA binding study demonstrated that evans blue (EB) has strong binding affinity with calf-thymus DNA (CT-DNA) (=5.08×10 L/(mol/L)). Furthermore, the accumulation of I-EB in necrotic muscle was efficiently blocked by an excess amount of unlabeled EB. In conclusion, I-EB can not only detect necrosis by binding the DNA released from necrotic cells, but also image necrotic tissues generated from the disease clinically.