Objective To investigate the protective effect of Xinyi capsule pretreatment on myocardial ischemia reperfusion injury in rabbits and its possible mechanism. Methods Ninety-four rabbits were randomly divided into 6 groups: model group (n=16), tirofiban group (n=16), high-, medium- and low-dose Xinyi capsule groups (4.0, 2.0, 1.0 g/kg;n=16 in each group), and sham operation group (n=14). Five days after intragastric administration with drug, myocardial ischemia reperfusion was induced by ligation of the proximal left circumflex artery. The electrocardiogram (ECG) was continuously recorded. The serum levels of myeloperoxidase (MPO), lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB) were measured. Myocardial histopathological damage was evaluated. Results The changes of J-point amplitude on ECG in high-, medium-and low-dose Xinyi capsule groups (0.064 ± 0.049 mV, 0.069 ± 0.061 mV, 0.079 ± 0.060 mV) were significantly lower than that in the model group (0.158 ± 0.105 mV, P<0.01 or P<0.05), the serum levels of LDH (399.7 ± 202.3 U/L, 369.6 ± 229.0 U/L, 435.5 ± 152.4 U/L), CK-MB (900.8 ± 231.2 U/L, 1 268.3 ± 899.8 U/L, 1 386.7 ± 621.6 U/L), MPO (69.81 ± 5.51 U/L, 85.44 ± 10.31 U/L, 81.33 ± 16.87 U/L) were significantly lower than those in the model group (LDH:817.1 ± 401.9 U/L, CK-MB:2 071.3 ± 693.5 U/L, MPO:149.9 ± 20.11 U/L;P<0.01 or P<0.05). Histopathological examination showed that myocardial damage in high-, medium- and low-dose Xinyi capsule groups reduced compared with the model group. Conclusions Xinyi capsule pretreatment can protect against myocardial ischemia reperfusion injury in rabbits, and its mechanism may be related to inflammation inhibition.

Acute pancreatitis (AP) is an inflammatory disease of the pancreas. Its pathogenesis is not only related to abnormal activation of trypsinogen, but also related to calcium overload, mitochondrial dysfunction, impaired autophagy and endoplasmic reticulum stress. However, the mechanism has not been fully elucidated and needs to be further studied. Currently, there is no effective treatment for AP. It is difficult to prevent the loss of pancreatic function. An in-depth understanding of the pathophysiological mechanisms of AP may help to identify the potential therapeutic targets. Therefore, the purpose of this study is to review recent advances in the mechanism of AP in order to provide more research direction for treatment.

Objective:To identify pathogenicity of the potential splicing variants in two Chinese Han patients with osteogenesis imperfecta.Methods:Genomic DNA was extracted using the conventional phenol-chloroform method; whole exome sequencing (WES) was used to analysis the disease-related variants in the two probands; Minigene assay was used to identify pathogenicity of the variants found in the patients' genome that possibly affect RNA splicing.Results:Two potential splicing variants, c.858+1_858+5delGTAAG in intron 12 of COL1A1 and c.1405-7C>T in intron 24 of COL1A2, were found in proband 1 and proband 2, respectively. In addition, a missense mutation, c.2972G>T (p.G991V) in exon 45 of COL1A2, was detected in proband 2. Minigene assay revealed that the variant in proband 1 caused the skipping of exon 12, while the variant in proband 2 did not lead to aberrant splicing. G199 of the COL1A2 in proband 2 was a highly conserved amino acid site, and the results suggested that c.2972G>T (p.G991V) may be the real pathogenic variant by the means of bioinformatics analysis.Conclusion:The variant c.858+1_858+5delGTAAG in COL1A1 was a causative variant that led to OI in proband 1, while the missense variant c.2972G>T (p.G991V) in COL1A2 was the cause of OI in proband 2, instead of the variant c.1405-7C>T. Minigene assay for potential splicing variants detected by WES could not only validate the pathogenicity of the candidate variants and enrich the mutation spectrum of OI, but also lay the foundation for patients' prenatal diagnosis and subsequent mechanism research.

Malignant tumors usually have no obvious clinical symptoms in the early stage. Most patients are already in the advanced stage when they are diagnosed. Some patients have lost the opportunity for operation, resulting in poor prognosis. Therefore, how to find the best therapeutic target for such patients and improve the prognosis of patients has gradually become the focus of scholar′s attention. Recently, Kruppel-like factor (KLF) is a transcriptional regulator that can bind to the target DNA, and its family plays an important role in the occurrence and development of malignant tumors. It has also been confirmed that the KLF family affects the proliferation, differentiation and migration of tumor cells, but the specific mechanism is still not fully elucidate. Consequently, in order to further explored the effect of the KLF family on tumors, this study intends to briefly review the roles and regulatory mechanisms of the KLF family in the cell proliferation, differentiation and migration of malignant tumors, hoping to provide new target for the biological treatment of tumors.

Exosomes are extracellular vesicles containing DNA, RNA and protein. They participate in intercellular communication and play an important role in tumor growth and metastasis. Exosomes exist in a variety of body fluids. Blood, urine and cerebrospinal fluid can be separated and extracted. Exosomes derived from tumor cells have the characteristics of tumor cells. Studies have shown that exosomes are involved in many processes of tumorigenesis and development, including information transmission between tumor cells, invasion and metastasis of tumor cells. Exosomes can also be used as carriers to deliver drugs to target cells, which has the potential of targeted therapy. In the process of tumor treatment, drugs can be designed based on the targeted recognition characteristics of exosomes, so as to improve the bioavailability of anti-tumor drugs, reduce adverse reactions and enhance the therapeutic effect. The research and application of exosomes are very challenging. There is a huge heterogeneity in the types, sizes and sources of exosomes, and the production mechanism is also very complex. This paper reviews the extraction and identification methods of exosomes, and reviews the clinical application of tumor derived exosomes.

An HPLC method for simultaneous determination of chlorogenic acid and mangiferin in original medicinal materials and decoction pieces of Pyrrosiae Folium was developed. The assay was performed on a Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column eluted with a mobile phase consisted of acetonitrile and 0.5% phosphoric acid solution in gradient elution at a flow rate of 1.0 mL x min(-1). The column temperature was set at 25 degrees C. The detection wavelength was 320 nm. The results showed that The linear ranges of chlorogenic acid and mangiferin were 5.2-130 mg x L(-1) (r = 0.9999) and 1.2-18 microg x mL(-1) (r = 0.9999), and the average recoveries (n=6) were 97.9% (RSD 1.9%) and 99.6% (RSD 2.9%), respectively. The method was simple, reproducible and valid. It can be used for quality evaluation and control of original medicinal materials and decoction pieces of Pyrrosiae Folium.
Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Xanthones ; analysis

Objective To develop an SPE‐HPLC method for the determination of methotrexate (M TX) in human plasma to monitor the clinical drug use of M TX ,and to identify the human metabolite of M TX by mass spectrometry .Methods M TX was extracted from human plasma using C18 SPE column and analyzed directly after elution .The separation of MTX was per‐formed on Diamonsil C18 column (150 mm × 4 .6 mm ,5 m) with a gradient mobile phase of methanol and 0 .5% acetic acid solu‐tion (containing 0 .3% triethylamine) at a flow rate of 1 .0 ml/min .The detection wavelength was 306 nm .QLMS and MRM‐IDA‐EPI scan modes were used to obtain the mass spectrum of MTX metabolite .Results The calibration curve of MTX in plasma was linear over the range of 0 .05‐100 μmol/L(r=0 .999 9) .The extraction recovery was above 95% and accuracy was between 97% and 105% .Both intra‐and inter‐day precision were less than 5% .7‐OH MTX was identified to be the metabolite through the analysis of the mass spectrum .Conclusion The method is sensitive ,reproducible ,easy to operate and suits for monitoring the concentration of M TX in clinical practice .

Objective:To analyze the effects of infectious complications [infected pancreatic necrosis (IPN) and extra-pancreatic infection (EPI)] on the outcomes of patients with severe acute pancreatitis (SAP), and evaluate the differences in infection time, infection site and infecting species between SAP patients with infections complications.Methods:The clinical data of 66 SAP patients with combined infectious complications admitted to Xuanwu Hospital, Capital Medical University from January 2014 to December 2020 were retrospectively analyzed, and SAP patients were divided into IPN group ( n=7), EPI group ( n=14) and co-infection (EPI+ IPN) group ( n=45) according to the type of infection. Whether the study data conformed to a normal distribution was assessed by the Shapiro-Wilk test, normally distributed measures were expressed as mean ± standard deviation ( ± s), and ANOVA was used for comparison between groups; skewed measures were expressed as median (interquartile range) [ M ( Q1, Q3)], and the rank-sum test was used for comparison between groups. Bonferroni correction was used for multiple group comparisons ( P value significance level reduced to 0.017). Quantitative data were compared between groups using the χ2 test or Fisher's exact probability method. Results:There were no statistical differences between the three groups in terms of baseline data at admission (gender, age, etiology, modified CTSI score, degree of pancreatic necrosis, and number of organ failure) ( P>0.05), patients in the EPI group were referred earlier than the other two groups ( P<0.05). In clinical treatment, patients in the IPN group and co-infection group required multiple minimally invasive interventions compared with those in the EPI group ( P<0.05), and the number of patients requiring combined nutritional support, length of intensive care unit stay, and total length of hospital stay were higher in the co-infection group than in the other two groups ( P<0.05). In addition, 360 strains of pathogenic bacteria were cultured in this study, with Gram-negative bacteria being the most common, and patients with SAP were more likely to have EPI in the early stage of disease onset, with bacteremia and respiratory tract infections in the early stage (≤14 d), and bacteremia, urinary tract infections, and catheter-associated infections in the late stage (>14 d). Conclusions:Among patients with SAP, patients in the co-infection group had higher surgical intervention, nutritional support and length of hospital stay than those in the single infection group. It is advisable to prioritize EPI in SAP patients with suspected infections, and the common infectious strains in SAP patients are still predominantly Gram-negative bacteria, and clinicians need to adjust the treatment plan in a timely manner according to the changes in patients′ conditions.

OBJECTIVE: To explore the pathogenic variants and clinical classification of two fetuses with Short-rib thoracic dysplasia with or without polydactyly (SRTD). METHODS: With informed consent obtained, the phenotypic characteristics of the fetuses were comprehensively examined, and genomic DNA was extracted from fetal skin tissue and peripheral blood samples of the parents with conventional phenol-chloroform method. Whole exome sequencing (WES) was carried out on both fetuses, and the candidate variants were validated by Sanger sequencing. The pathogenicity of the candidate variants was analyzed using bioinformatic software VarCards, and the impact of the variants on the protein structure was predicted with Swiss-Pdb-viewer. RESULTS: Both fetuses were found to harbor compound heterozygous variants of the DYNC2H1 gene, including c.515C>A (p.Pro172Gln) and c.5983G>A (p.Ala1995Thr) in fetus 1, and c.5920G>T (pGly1974) and c.9908T>C (p.He3303Thr) in fetus 2. The parents of both fetuses were heterozygous carriers. CONCLUSION The compound heterozygous variants of the DYNC2H1 gene probably underlay the SRTD3 in the two fetuses.
Humans ; Fetus ; Chloroform ; Computational Biology ; Ethnicity ; Ribs

Acute and chronic wounds seriously threaten patients' life health and quality of life, therefore, wound repair has become a hot topic of research for scholars at home and abroad in recent years. With the development of material science and tissue engineering, more and more biomaterials prepared from natural ingredients were used in basic research and clinical treatment of wound repair. Such biomaterials can be used as templates for wound tissue regeneration to induce autologous cell adhesion and migration, and promote the deposition of extracellular matrix, which have broad clinical application prospects. This paper reviews the characteristics and application advance of natural biomaterials which are popular in the field of wound repair, aiming to provide ideas for the research and development of new wound dressing and tissue engineering skin.