Expression of LRG-1 in mice with hypertensive renal damage and its significance / 中南大学学报(医学版)

OBJECTIVES@#Long-term elevated blood pressure may lead to kidney damage, yet the pathogenesis of hypertensive kidney damage is still unclear. This study aims to explore the role and significance of leucine-rich alpha-2-glycoprotein-1 (LRG-1) in hypertensive renal damage through detecting the levels of LRG-1 in the serum and kidney of mice with hypertensive renal damage and its relationship with related indexes.@*METHODS@#C57BL/6 mice were used in this study and randomly divided into a control group, an angiotensin II (Ang II) group, and an Ang II+irbesartan group. The control group was gavaged with physiological saline. The Ang II group was pumped subcutaneously at a rate of 1.5 mg/(kg·d) for 28 days to establish the hypertensive renal damage model in mice, and then gavaged with equivalent physiological saline. The Ang II+irbesartan group used the same method to establish the hypertensive renal damage model, and then was gavaged with irbesartan. Immunohistochemistry and Western blotting were used to detect the expression of LRG-1 and fibrosis-related indicators (collagen I and fibronectin) in renal tissues. ELISA was used to evaluate the level of serum LRG-1 and inflammatory cytokines in mice. The urinary protein-creatinine ratio and renal function were determined, and correlation analysis was conducted.@*RESULTS@#Compared with the control group, the levels of serum LRG-1, the expression of LRG-1 protein, collagen I, and fibronectin in kidney in the Ang II group were increased (all P<0.01). After treating with irbesartan, renal damage of hypertensive mice was alleviated, while the levels of LRG-1 in serum and kidney were decreased, and the expression of collagen I and fibronectin was down-regulated (all P<0.01). Correlation analysis showed that the level of serum LRG-1 was positively correlated with urinary protein-creatinine ratio, blood urea nitrogen, and blood creatinine level in hypertensive kidney damage mice. Serum level of LRG-1 was also positively correlated with serum inflammatory factors including TNF-α, IL-1β, and IL-6.@*CONCLUSIONS@#Hypertensive renal damage mice display elevated expression of LRG-1 in serum and kidney, and irbesartan can reduce the expression of LRG-1 while alleviating renal damage. The level of serum LRG-1 is positively correlated with the degree of hypertensive renal damage, suggesting that it may participate in the occurrence and development of hypertensive renal damage.

Single-cell RNA sequencing reveals the transcriptomic landscape of kidneys in patients with ischemic acute kidney injury / 中华医学杂志(英文版)

BACKGROUND@#Ischemic acute kidney injury (AKI) is a common syndrome associated with considerable mortality and healthcare costs. Up to now, the underlying pathogenesis of ischemic AKI remains incompletely understood, and specific strategies for early diagnosis and treatment of ischemic AKI are still lacking. Here, this study aimed to define the transcriptomic landscape of AKI patients through single-cell RNA sequencing (scRNA-seq) analysis in kidneys.@*METHODS@#In this study, scRNA-seq technology was applied to kidneys from two ischemic AKI patients, and three human public scRNA-seq datasets were collected as controls. Differentially expressed genes (DEGs) and cell clusters of kidneys were determined. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, as well as the ligand-receptor interaction between cells, were performed. We also validated several DEGs expression in kidneys from human ischemic AKI and ischemia/reperfusion (I/R) injury induced AKI mice through immunohistochemistry staining.@*RESULTS@#15 distinct cell clusters were determined in kidney from subjects of ischemic AKI and control. The injured proximal tubules (PT) displayed a proapoptotic and proinflammatory phenotype. PT cells of ischemic AKI had up-regulation of novel pro-apoptotic genes including USP47 , RASSF4 , EBAG9 , IER3 , SASH1 , SEPTIN7 , and NUB1 , which have not been reported in ischemic AKI previously. Several hub genes were validated in kidneys from human AKI and renal I/R injury mice, respectively. Furthermore, PT highly expressed DEGs enriched in endoplasmic reticulum stress, autophagy, and retinoic acid-inducible gene I (RIG-I) signaling. DEGs overexpressed in other tubular cells were primarily enriched in nucleotide-binding and oligomerization domain (NOD)-like receptor signaling, estrogen signaling, interleukin (IL)-12 signaling, and IL-17 signaling. Overexpressed genes in kidney-resident immune cells including macrophages, natural killer T (NKT) cells, monocytes, and dendritic cells were associated with leukocyte activation, chemotaxis, cell adhesion, and complement activation. In addition, the ligand-receptor interactions analysis revealed prominent communications between macrophages and monocytes with other cells in the process of ischemic AKI.@*CONCLUSION@#Together, this study reveals distinct cell-specific transcriptomic atlas of kidney in ischemic AKI patients, altered signaling pathways, and potential cell-cell crosstalk in the development of AKI. These data reveal new insights into the pathogenesis and potential therapeutic strategies in ischemic AKI.

Modern Research Progress of Polyphyllin Ⅰ： A Review / 中国实验方剂学杂志

Paridis Rhizoma， a traditional valuable Chinese herbal medicine， has the functions of clearing heat and removing toxin， relieving edema and pain， cooling liver and calming convulsion， which can be used to treat various diseases such as mumps， abscess， burn， bleeding， and tumor. It has been used in folk medicine for a long time and is the main raw material of various Chinese patent medicines such as Gongxuening Capsules and Yunnan Baiyao. Polyphyllin Ⅰ， an isospirostanol saponin and one of the main active components in Paridis Rhizoma， is distributed in the rhizome， pericarp， and leaves of Paris polyphylla. With high polarity， polyphyllin Ⅰ is mainly extracted by n-butanol extraction and macroporous adsorption resin chromatography， separated by silica gel column chromatography and preparative high performance liquid chromatography， and purified with the combination of methods. With anti-tumor， anti-inflammatory， antibacterial， and anti-virus effects， it is generally employed to treat liver cancer， lung cancer， gastric cancer and other cancers as well as arthritis， influenza， sore toxin， and bacterial infection. However， polyphyllin Ⅰ may cause stomach irritation， hemolysis， liver damage， kidney damage， heart damage， and other adverse reactions. Pharmacokinetic studies show that it has problems such as low bioavailability and poor intestinal absorption and permeability， which affect the clinical application of polyphyllin Ⅰ. This paper summarizes the research on the plant sources， extraction and separation methods， pharmacological effects， adverse reactions， and pharmacokinetics of polyphyllin Ⅰ in recent years， which is expected to provide a reference for the rational clinical application and other in-depth research work of polyphyllin Ⅰ.

Clinical Effects of Trimetazidine Combined with Salvia Ligustrazine Injections in the Treatment of Unstable Angina in Coronary Heart Disease with Type 2 Diabetes / 中国药师

Objective:To investigate the clinical efficacy of trimetazidine combined with salvia ligustrazine injections in the treat-ment of unstable angina in coronary heart disease with type 2 diabetes. Methods:Totally 76 patients of unstable angina with type 2 di-abetes were randomly divided into the control group and the observation group with 38 cases in each. The control group received trime-tazidine on the basis of the conventional treatment, 20mg,po,tid, while the patients in the observation group received salvia ligustrazine injections additionally, 10ml, ivd,qd. The treatment course was 2 weeks, and the clinical efficacy, blood fat, blood rheology and the other indices were observed after the treatment. Results:After the treatment, the effective rate of the control group and the observation group was 65. 79% and 86. 84%,respectively with statistically significant difference(P<0. 05), the improvement of blood fat and blood rheology in the observation group was better than that in the control group (P<0. 05), the angina frequency and duration, and nitroglycerin consumption in the observation group was significantly lower than those in the control group(P<0. 05). Conclusion:The combination application containing trimetazidine and salvia ligustrazine shows exact clinical efficacy with promising safety and compli-ance, which is valuable in the clinical application.

Construction and expression of recombinant retrovirus vector carrying HBV vector / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To explore the possibility of using retrovirus vector to carry HBV vector, and to prove that replication defective HBV could be normally packaged.</p><p><b>METHODS</b>Two kinds of full length of mutant HBV gene, which express dominant negative mutants, were inserted into retrovirus vector. After recombinant retroviruses were harvested, they were used to infect Hep G2 and 2.2.15 cell line. Then the expression of HBV core antigen in the Hep G2 cell was examined by immune fluorescence, and the existence of recombinant HB virion in the culture medium was examined by PCR.</p><p><b>RESULTS</b>High titer of recombinant retroviruses were obtained in the culture medium of transfected PA317 cell line. Core antigen was detectable in the recombinant retrovirus infected Hep G2 cell. Recombinant HB virion was detectable in the culture medium of recombinant retrovirus infected 2.2.15 cell.</p><p><b>CONCLUSIONS</b>The results suggested that recombinant retrovirus could carry HBV vector and express HBV products. When structural protein is offered by wt-HBV, the recombinant retrovirus may function as HBV vector, therefore it could be used in anti?HBV gene therapy.</p>

Study on the antiviral effect of C gene truncated HBV mutant in HBV infection / 中华传染病杂志

Objective To explore the effect of the C gene truncated HBV mutant on HBV replication. Methods The C gene truncated HBV vector pHBV-?C was constructed through the molecular clone in vitro,and then transfected transiently into HepG2 cell.The expression of S protein was assayed by ELISA and SDS-PAGE Western blot.After co-transfection with pHBV-?C and wild HBV genome adwR9 into HepG2 cell the DNA was detected quantitatively by real-tine fluorescence quantitative PCR in the culture medium and the cell. Results There was no significant difference in expression of S protein assay by ELISA and Western blot.The DNA of the cotransfected group with pHBV-?C and adwR9 was lower than that of control group in the culture medium and the cell. Conclusions(C gene) truncated HBV mutant can cause the reduction of HBV replication.

DNA mediated immunization co-injected with CpG-oligodeoxynucleotides in hepatitis B virus transgenic mice / 中国免疫学杂志

Objective:To study the immune responses of HBV based immunization co injecting CpG oligodeoxynucleotides in HBV transgenic mice.Methods:The HBV transgenic mice were immunized by multiple sites intramuscular injection with V HBs and CpG ODN or V HBs alone or V 1012 controls.The anti HBs Ab and HBsAg in serum from immunized mice was measured by ELISA.The expression of HBsAg in liver tissue was detected by immunohistochemistry method(SP).The histological activity index was calculated with Knodell's method.Results:The anti HBs Ab in serum could be detected and HBsAg in serum and liver tissues could not be measured in 2 out of 6 HBV transgenic mice immunized with V HBs+CpG ODN.There were not significantly changes in the level of anti HBs Ab and HBsAg in serum from HBV transgenic mice immunized with V HBs or V 1012 alone.The histological activity indexes in liver tissues from mice immunized with V HBs+CpG ODN were higher than that in mice inoculated with V HBs or V 1012 alone.The large number of lymphocytes could be found in liver tissues in V HBs+CpG ODN immunization group by microscopy.Conclusion:The immuno response,which seems to be responsible for the disappearance of HBsAg,might be elicited in HBV transgenic mice immunize with V HBs+CpG ODN.