Primary biliary cholangitis (PBC) is an autoimmune liver disease with unclear pathogenesis.The amino acid composition and sequence in the complementarity-determining region 3 of T cell receptor (TCR) and B cell receptor (BCR) are highly diverse, which forms a large antigen recognition receptor repertoire, i.e., immune repertoire.In recent years, second-generation sequencing techniques combined with multiplex PCR or amplicon rescue multiplex PCR have been used to study the features of immune repertoire in PBC patients, and it has been found that PBC patients have clonal expansion of specific CD4+ T lymphocytes, clonal diversity of B lymphocytes, somatic hypermutation, and reduction in class switch, as well as increase in clonal diversity after treatment with ursodeoxycholic acid.These findings need to be confirmed by large-scale in vivo and in vitro studies and different immune repertoire research strategies.

BACKGROUND:In recent years, some studies have demonstrated that ganglioside can promote survival and differentiation of umbilical blood cord mesenchymal stem cel s in vitro. OBJECTIVE:To observe the effect of injection of human umbilical blood cord mesenchymal stem cel s and ganglioside into rat lateral ventricles on neurological functional recovery from cerebral palsy. METHODS:Total y 60 cerebral palsy neonatal rats were delivered from pregnant rats which were modes were given intraperitoneal injection of lipopolysaccharide for 2 successive days on day 17 of gestation. Then those neonatal rats were randomly divided into five groups, including model group (n=10), sham transplantation group (n=10), stem cel transplantation group (n=18), ganglioside group (n=10) and combination group (n=12). Under stereotaxic instrument, umbilical blood cord mesenchymal stem cel s or ganglioside were injected into left lateral ventricles of the rat brain, respectively, and the sham transplantation group was given the same volume of phosphate buffered saline. Two rats from the stem cel transplantation group were put to death for immunofluorescence staining at 7, 14, 21 and 28 days after transplantation, respectively, and two rats in the combination group were kil ed for immunofluorescence staining at 14 days. Besides, al rats were underwent neurologic evaluation at 28 days after transplantation. RESULTS AND CONCLUSION:The umbilical blood cord mesenchymal stem cel s could survive, migrate and differentiate, which mainly distributed in the lateral ventricle, hippocampus and cortex. At 14 days after transplantation, positive expressions of BrdU and glial fibril ary acidic protein in the combination group were significantly higher than those in the stem cel transplantation group (P<0.05). In addition, compared with the model group, the holding time significantly prolonged and foot error times significantly decreased in the latter three groups (P<0.05), as wel as in the combination group compared with the stem cel transplantation and ganglioside groups (P<0.05). These results indicate that umbilical blood cord mesenchymal stem cel s and ganglioside can both improve neurological function of rats with cerebral palsy. Given that ganglioside can promote survival and differentiation of umbilical blood cord mesenchymal stem cel s in vivo, the combined transplantation is preferred.

BACKGROUND: The uterine arterial embolization which is a major method to treat hysteromyoma has bean widely used in clinic and achieved a satisfactory therapeutic efficacy. The study addressing the effect of trisacryl gelatin microspheres on uterine arterial embolization in a hysteromyoma guinea pig model has less bean reported yet. OBJECTIVE: To vedfy the feesibility of trisacryl gelatin microspheres to uterine arterial embolization in hysteromyoma guinea pig models. METHODS: A total of 30 adult female guinea pigs were randomly divided into two groups: pelvic cavity artery moulding group (n=10) was performed pelvic vascular casting mould to demonstrate the anatomical characteristics, such as source, running shape, length, diameter and branches; arterial embolization group (n=20) was induced hysteromyoma model using astrogen-progestogen replacement therapy and performed technical research and pathological analysis by bilateral uterine arterial embolization. RESULTS AND CONCLUSION: The trunks of uterine arteries were erupted from internal iliac arteries. The diameter of the trunks and its arcuate branches were (0.350±0.022) mm and (0.160±0.012) mm, respectively. The 20 guinea pigs of the arterial embolization group were succeeded in operating bilateral arterial embolization. The dosage of 40-120 pm and 100-300 μm trisacryl gelatin microspheras were (0.040t±0.005) mL and (0.017±0.002) mL respectively during the operation. The achievement ratio of establishing model was 75% in the arterial embolization group. On the pathological section, the microspheres could be found in the uterine arterial arcuate branches and second branches within the subsercsa and third branches. The myometrium Was thickening. The cells of the leiomyoma nodules arranged in palisade or weaving shapes. Ischemia and necrosis were evidently present in leiomyomas of guinea pigs after embolization, but the myometria and endometria had no pathological change of ischemia and necrosis. It is feasible to use trisacryl gelatin microspheres to operate uterine arterial embolization for hysteromyoma of guinea pigs and the embolization effects are satisfactory.

[Objective]This study was designed to evaluate the effects of percutaneously puncturing vertebrate adjacent to cartilage endplate and injecting pingyangmycin on lumbar intervertebral disc and cartilage endplate in New Zealand Rabbits.[Methods]Thirty-six New Zealand white rabbits were enrolled in this study.The fifth lumbar vertebmte(L_5)was injected with pingyangrnycin as experimental group,and the fourth lumbar vertebmte(L_4)injected normal sodium as control group.Six rabbits were selected randomly,then MRI and histological observation was performed in the first,second,third,fourth,Fifth week and third month after operation respectively.Moreover,the correlation analysis was performed between MRI and histological measurements for areas of the lesion in L_5.[Results]There was no obvious changes on MRI and histological examination in control group.For experimental group,there were also no obvious changes in the first two weeks after bperation.However,in the third week,it demonstrated slightly hyperintense signal on T_2WI and fat-suppression T_2WI(FS T_2WI),while FS T_1WI was hypointense signal.The signal changed more obviously in the fourth week.Histologically,the structure of vertibrates arranged disordedy,chondrocyte of endplate decreased and architecture became disorder.Anulus fibrosus and nucleus pulposus did not change.The cartilage endplate and intervertebral disc degenerated in the fifth week.Both of them degenerated more obviously in third month.There was a strong correlation between MRI and histological measurements for areas of the lesion in the fourth week(r=0.965,P＜ 0.001).[Conclusion]Degeneration of lumbar intervertebral disc and cartilage endplate in New Zealand Rabbits can be induced by percutaneously puncturing vertebrate adjacent to cartilage endplate and injecting pingyangmycin.

Objective To analyze the genetic quality of 24 domestic inbred strains mice using microsatellite loci panel.Methods Previously selected 30 microsatellite loci of mouse with high polymorphism and more allele numbers were used to synthesize corresponding fluorescently-labeled primers.Then the genomic DNA samples of each mouse were amplified by PCR and the products were analyzed by STR scanning to genotype the inbred strains of mice.Results Out of the 24 inbred strains, 15 inbred strains showed the same genotype within one strain at 30 loci.Among different strains, microsatellite loci indicated polymorphism which could be used to distinguish different strains.However, the rest 9 strains demonstrated polymorphism within strains.Conclusions Our stuoly provides a useful microsatellite panel to detect genetic quality of inbred mice and distinguish different strains with the optimized microsatellite loci.

With the rapid development of transgenic technology, the safety of genetically modified products has received extensive attention. Certified reference materials for the detection of genetically modified organisms play important roles in ensuring comparability and traceability of the qualitative and quantitative detection of genetically modified products. However, the development of protein reference materials is relatively slow, and one of the difficulties is the preparation of protein candidates with high purity. The cry1Ah1 gene of Bacillus thuringiensis has been used for the development of transgenic insect-resistant crops because of its excellent insecticidal activity against lepidopteran pests such as Asian corn borer, and has obtained transgenic lines with good insect resistance traits. In order to develop Cry1Ah protein certified reference material, it is urgent to establish a preparation and purification system. In this study, a system for preparing Cry1Ah protein by Bt expression system was optimized, and a high-purity Cry1Ah protein (size exclusion chromatography purity: 99.6%) was obtained by ion-exchange chromatography and size exclusion chromatography stepwise purification. The results of biological activity assay showed that there was no significant difference in the insecticidal activity of purified Cry1Ah protein and protoxin against diamondback moths (Plutella xylostella). Finally, the amino acid sequence of the activated Cry1Ah protein was determined using Edman degradation and mass spectrometry. In summary, the obtained Cry1Ah pure protein can be used for the development of protein reference materials.
Animals ; Bacillus thuringiensis ; Bacterial Proteins ; Cryptochromes ; metabolism ; Endotoxins ; Hemolysin Proteins ; Moths ; Pest Control, Biological ; Plants, Genetically Modified

Objective To investigate the clinical efficacy of low-frequency transcranial magnetic stimulation (rTMS) on post-stroke upper limb spasticity and its mechanism. Methods From September, 2015 to December, 2017, 23 patients with post-stroke upper limb paralysis were randomly divided into control group (n=13) and experimental group (n=10). Both groups received routine rehabilitation, and the experimental group received 1 Hz rTMS at primary motor area (M1) for eight weeks. They were assessed with modified Ashworth Scale (MAS), modified Barthel Index (MBI) and Fugl-Meyer Assessment-Upper Extremities (FMA-UE) before and after treatment, while the activation under fMRI in the task state was observed and the laterality index (LI) was calculated. Results The scores of MAS, FMA-UE and MBI improved after treatment in both groups (Z>2.121, t=6.248, P<0.05), and improved more in the experimental group than in the control group (Z>2.084, t=-2.095, P<0.05). The ipsilateral M1, ipsilateral sensory motor cortex and bilateral supplementary motor area were activated more in the control group than in the experimental group during the movement of affected hand. LI in the M1 increased after treatment in both groups (Z>2.366, P<0.05), and was more in the experimental group than in the control group (Z=-2.430, P<0.05). There was a positive correlation between the change of LI in the M1 and the improvement of the MAS and FMA-UE (r>0.612, P<0.05). Conclusion Low-frequency rTMS may improve the motor function and spasticity of upper limb after stroke by promoting reorganization of the cortex and inducing normalization of cortical function.

Objective: To analyze the characteristics of immunoglobulin heavy chain complementarity-determining region (IgH-CDR3) repertoire of peripheral B cells in a patient with primary biliary cholangitis (PBC) and to investigate the diversity of the immune system. Methods: Arm-PCR was used to amplify the IgH-CDR3 region of circulating B cells isolated from a PBC patient, and high-throughput sequencing was used to analyze the amplified product. The characteristics of immune repertoire were analyzed by bioinformatics. Results: In total, 329219 sequence reads were generated from the sample, with 325540 total CDR3 sequences and 72774 distinct CDR3 sequences, and the D50 of IGH-CDR3 was 7.7. The dominant CDR3 length of the sample was 45 nt (9.6%); the N addition with the highest frequency ranged from 13 to 14 nt (5.25%); the J trimming with the highest frequency was 0 nt (12.7%); the three most frequent V alleles were V4-59 (9.5%), V3-23 (8.1%), and V1-69 (6.4%). Conclusion The diversity of IgH-CDR3 repertoire is relatively low in this patient with PBC, with several B-cell clonal expansions. The specificity needs to be further verified after increasing the sample size.

Objective:The investigation and research on the application status of Hepatic Venous Pressure Gradient (HVPG) is very important to understand the real situation and future development of this technology in China.Methods:This study comprehensively investigated the basic situation of HVPG technology in China, including hospital distribution, hospital level, annual number of cases, catheters used, average cost, indications and existing problems.Results:According to the survey, there were 70 hospitals in China carrying out HVPG technology in 2021, distributed in 28 provinces (autonomous regions and municipalities directly under the central Government). A total of 4 398 cases of HVPG were performed in all the surveyed hospitals in 2021, of which 2 291 cases (52.1%) were tested by HVPG alone. The average cost of HVPG detection was (5 617.2±2 079.4) yuan. 96.3% of the teams completed HVPG detection with balloon method, and most of the teams used thrombectomy balloon catheter (80.3%).Conclusion:Through this investigation, the status of domestic clinical application of HVPG has been clarified, and it has been confirmed that many domestic medical institutions have mastered this technology, but it still needs to continue to promote and popularize HVPG technology in the future.