Cloning and Function Identification of a Phytoene Desaturase Gene from Eucommia ulmoides

The phytoene desaturase(PDS)encodes a crucial enzyme in the carotenoid biosynthesis pathway.Silencing or inhibiting PDS expression leads to the appearance of mottled,chlorosis,or albino leaves.In this study,the CDS sequence of EuPDS(Eucommia ulmoides Phytoene Desaturase)was first cloned and then PDS was silenced in Nicotiana benthamiana.Result showed the expression level of EuPDS in leaves was higher than that in the roots and stems.In N.benthamiana leaves,which were treated by Agrobacterium for 24 h,photo-bleaching was shown on the fresh leaves one week after injection and the transcript level of PDS was down-regulated during the period of emersion.This suggested that EuPDS could silence PDS of N.benthamiana,so as to cause the phenotype of leaf whitening.PDS is the main reporter gene involved in virus-induced gene silencing(VIGS).This study offered molecular evidence for identifying PDS gene involved in Carotenoid’s biosynthesis pathway and the regulation networks in E.ulmides.It also laid a useful foundation for study on leaf discoloration mechanism of other woody plants.

Precore mutation enhances viral replication to facilitate persistent infection especially in HBeAg-negative patients

Naturally occurred precore(PC,G1896A)and/or basal core promoter(BCP,A1762T/G1764A)mutations are prevalent in chronic HBV-infected patients,especially those under HBeAg-negative status.However,the replicative capacity of HBV with PC/BCP mutations remains ambiguous.Herein,meta-analysis showed that,only under HBeAg-negative status,the serum HBV DNA load in patients with PC mutation was 7.41-fold higher than those without the mutation.Both PC mutation alone and BCPþPC mutations promoted HBV replication in cell and hydrodynamic injection mouse models.In human hepatocyte chimeric mouse model,BCPþPC mutations led to elevated replicative capacity and intrahepatic core protein accumulation.Mechanistically,preC RNA harboring PC mutation could serve as mRNA to express core and P proteins,and such pgRNA-like function favored the maintenance of cccDNA pool under HBeAg-negative status.Additionally,BCPþPC mutations induced more extensive and severe human hepatocyte damage as well as activated endoplasmic reticulum stress and TNF signaling pathway in livers of chimeric mice.This study indicates that HBeAg-negative patients should be monitored on HBV mutations regularly and are expected to receive early antiviral treatment to prevent disease progression.

Blood pressure targets in patients with chronic kidney disease:Comments and controversies

According to the Global Burden of Disease Study 2017,there are 132 million patients with chronic kidney disease(CKD)in China,^([1])and the prevalence of hypertension in Chinese patients with CKD(67.3%)is significantly higher than that in the general population(23.2%).^([2,3])CKD is both a cause and a consequence of hypertension,and these two conditions form a vicious cycle that leads to the high prevalence rate of hypertension in patients with CKD.Uncontrolled hypertension accelerates the deterioration of renal function,which significantly increases the risk of developing end-stage renal disease(ESRD),as well as cardiovascular and cerebrovascular diseases,leading to poor patient outcomes.

Comparative risk of acute kidney injury among cancer patients treated with immune checkpoint inhibitors

With the development and introduction of immune checkpoint inhibitors(ICIs)in cancer patients,immune-related side effects have increasingly attracted attention.However,the risks of immune-related renal toxicity are poorly characterized.In this study,we performed a network meta-analysis(NMA)of ICI-related randomized clinical trials(RCTs)to elucidate the comparative risk of acute kidney injury(AKI)in cancer patients receiving different ICIs.We also sought to identify other factors potentially affecting the risk of AKI.PubMed and EMBASE were searched for peer-reviewed trial reports published between January 2000 and May 2021.Eligible studies were RCTs studying ICIs in cancer patients and reporting AKI data.We performed a frequentist NMA to evaluate the risk ratios for grade 1-5 and grade 3-5 AKI between the treatment groups.We also assessed the absolute incidence of AKI in the ICI-containing arm using traditional direct meta-analysis.Once significant heterogeneity was detected in a traditional direct meta-analysis,multivariable meta-regression analysis was applied to identify factors that significantly affected the absolute incidence of AKI.A total of 85 RCTs were included in this study.In the NMA for the risk of grade 1-5 and 3-5 AKI,ipilimumab showed a significantly higher risk than avelumab and durvalumab,whereas 1 mg/kg nivolumab plus 3 mg/kg ipilimumab(N1I3)showed a significantly higher risk than other groups.In terms of treatment ranking,durvalumab±low-dose tremelimumab and avelumab were consistently among the top three safest treatments for grade 1-5 or 3-5 AKI,whereas N1I3,ipilimumab and tremelimumab were consistently among the top three treatments with the highest risk for grade 1-5 or 3-5 AKI.Compared with other cancers,renal cell carcinoma and urothelial carcinoma showed a significantly higher risk of AKI.The incidence of AKI was significantly higher with ICI+chemotherapy than with ICI monotherapy.In this NMA involving largescale up-to-date ICI trials,we demonstrated the comparative safety of existing ICI drugs for grade 1-5 and grade 3-5 AKI.Based on data from the ICI arms of these trials,we also revealed several potential risk factors for immune-related AKI,including tumor type and treatment paradigm.

RNA binding protein TIAR modulates HBV replication by tipping the balance of pgRNA translation

The pregenomic RNA(pgRNA)of hepatitis B virus(HBV)serves not only as a bicistronic message RNA to translate core protein(Cp)and DNA polymerase(Pol),but also as the template for reverse transcriptional replication of viral DNA upon packaging into nucleocapsid.Although it is well known that pgRNA translates much more Cp than Pol,the molecular mechanism underlying the regulation of Cp and Pol translation efficiency from pgRNA remains elusive.

关于近期不明原因儿童严重急性肝炎可能病因及发病机制与应对策略的探讨

2022年4月以来, 不明原因儿童严重急性肝炎已蔓延全球35个国家和地区, 共计1 010余例病例被相继报道。此次肝炎涉及地域较广且重症率较高, 因此尽早锁定病因并建立有效的预防、诊断和治疗措施至关重要。本文讨论了近期不明原因儿童严重急性肝炎的可能病因和发病机制, 并推测该病的发生可能与腺病毒、腺相关病毒感染及新型冠状病毒流行有关, 而不同种族间HLA多态性的差异则可能与报告病例多见于欧美国家有关。根据目前的相关证据, 可初步判定我国大规模暴发不明原因儿童严重急性肝炎的风险较低, 但我们仍需提高对该病的认识和警惕, 以积极准备应对策略。

Non-genetic mechanisms of diabetic nephropathy

Diabetic nephropathy （DN） is one of the most common microvascular complications in diabetes mellitus patients and is characterized by thickened glomeruIar basement membrane, increased extracellular matrix formation, and podocyte loss. These phenomena lead to proteinuria and altered glomerular filtration rate, that is, the rate initially increases but progressively decreases. DN has become the leading cause of end-stage renal disease. Its prevalence shows a rapid growth trend and causes heavy social and economic burden in many countries. However, this disease is multifactorial, and its mechanism is poorly understood due to the complex pathogenesis of DN. In this review, we highlight the new molecular insights about the pathogenesis of DN from the aspects of immune inflammation response, epithelial-mesenchymal transition, apoptosis and mitochondrial damage, epigenetics, and podocyte-endothelial communication. This work offers groundwork for understanding the initiation and progression of DN, as well as provides ideas for developing new prevention and treatment measures.

Potential use of serum HBV RNA in antiviral therapy for chronic hepatitis B in the era of nucleos（t）ide analogs

Although the efficacy of nucleos（t）ide analogue （NA） has been confirmed for treatment of chronic hepatitis B, long-term therapy has been recommended due to the high frequency of off-therapy viral DNA rebound and disease relapse. In this review, the RNA virion-like particles of hepatitis B virus （HBV） are integrated into the Hfe cycle of HBV replication, and the potential significance of serum HBV RNA is systematically described. The production of HBV RNA virion-like particles should not be blocked by NA; in this regard, serum HBV RNA is found to be a suitable surrogate marker for the activity of intrahepatic covalently closed circular DNA （cccDNA）, particularly among patients receiving NA therapy. Therefore, the concept of virological response is redefined as persistent loss of serum HBV DNA and HBV RNA. In contrast to hepatitis B surface antigen （HBsAg） that can originate from either the cccDNA or the integrated HBV DNA fragment, serum HBV RNA, with pregenomic RNA origination, can only be transcribed from cccDNA. Therefore, the loss of serum HBV RNA would likely be a promising predicator for safe drug discontinuation. The clinical status of consistent loss of serum HBV RNA accompanied with low serum HBsAg levels might be implicated as a ＂para-functional cure,＂ a status nearly close to the functional cure of chronic hepatitis B, to distinguish the ＂functional cure＂ characterized as serum HBsAg loss with or without anti-HBs seroconversion.

MXene介导的近红外光热效应用于快速清除多种耐药菌以及菌膜

随着细菌耐药性的不断出现和加重,针对耐药菌感染和菌膜这类难以用传统抗生素治疗的临床医疗问题,需要发展新型高效快速的杀菌方法.本文采用新型二维材料MXene与近红外激光相结合,实现了在20 min内对细菌以及菌膜的快速高效杀除.为了测试该方案的广谱抗菌性,我们对包括耐药性的耐甲氧西林金黄色葡萄球菌(MRSA)和耐万古霉素肠球菌(VRE)等多种耐药菌进行了快速灭菌实验.我们发现二碳化三钛类的MXene与808 nm激光的组合,对所测试的15种细菌均展现出显著的抗菌特性.并且该杀菌方案可通过破坏菌膜结构杀灭深层细菌,快速清除由MRSA所形成的菌膜.此外,对抗菌机理的研究显示,MXene和近红外激光主要是通过物理性的MXene插入及接触和光热效应杀死细菌,从而可以显著地降低细菌耐药性的产生.本工作提出了一种物理性清除多种耐药菌和菌膜的方案,并扩展了新型二维材料MXene在生物医学领域和临床上的潜在应用范围.

Integration of Prealbumin into Child-Pugh Classification Improves Prognosis Predicting Accuracy in HCC Patients Considering Curative Surgery

Background and Aims:The poor outcomes of hepatocellular carcinoma(HCC)patients may be due to not only malignant tumors but also limited liver function.Therefore,as stated in major guidelines,only patients with relatively normal liver function(Child-Pugh A)would be referred for curative hepatectomy.Even so,the postsurgery survival rate of patients is still extremely poor.Direct curative resection may benefit most patients.This study aimed to improve the prognosis predicting accuracy of the Child-Pugh scoring system.Methods:This study included two cohorts:cohort A being composed of 613 HCC patients,with a 23-month median postsurgery follow-up time;and cohort B being composed of 554 tumor-free chronic liver disease patients.Kaplan-Meier test and Cox model were used for survival analysis.Independent-samples t test or one-way ANOVA was used to test the differences between different groups.Results:Serum prealbumin levels were found inversely correlated with worsening of flbrotic scores(r=-0.482,p<0.001).Lower levels of presurgery prealbumin was an independent factor of poor postsurgery prognosis in Child-Pugh A patients,with a hazard ratio of 0.731(p=0.001).By integrating prealbumin together with total bilirubin level,serum albumin concentration and prothrombin time,a modified liver disease prognosis scoring system was developed to define traditional Child-Pugh A HCC patients as Modified Child-Pugh MCP-1,MCP-2 and MCP-3,with median postsurgery overall survival times of 44.00,28.00 and 11.00 months respectively.Conclusions:Preoperative serum prealbumin is a valuable prognosis predicting biomarker for Child-Pugh A HCC patients who may be under consideration for curative resection.With serum prealbumin included as one of the parameters,the MCP scoring system might improve the postsurgery survival predicting accuracy for HCC patients.

Netrin-1 works with UNC5B to regulate angiogenesis in diabetic kidney disease

Netrin-1,an axon guidance factor,and its receptor UNC5B play important roles in axonal development and angiogenesis.This study examined netrin-1 and UNC5B expression in kidneys with diabetic kidney disease (DKD) and investigated their roles in angiogenesis.Netrin-1 and UNC5B were upregulated in streptozotocininduced DKD Wistar rats,and their expression was compared with that in healthy controls.However,exogenous netrin-1 in UNC5B-depleted human renal glomerular endothelial cells (HRGECs) inhibited cell migration and tubulogenesis.This effect was likely associated with SRC pathway deactivation.Netrin-1 treatment also eliminated the pro-angiogenic effects of exogenous VEGF-165 on UNC5B-silenced HRGECs.These results indicate that UNC5B antagonizes netrin-1 and that UNC5B upregulation contributes partly to enhancing angiogenesis in DKD.Therefore,introducing exogenous netrin-1 and depleting endogenous UNC5B are potential strategies for reducing the incidence of early angiogenesis and mitigating kidney injury in DKD.

The role of transcriptional factor D-site-binding protein in circadian CCL2 gene expression in anti-Thy1 nephritis

Mesangial proliferative glomerulonephritis(MsPGN)is an inflammatory disease,but both the nature of disease progression and its regulation remain unclear.In the present study,we monitored the course of anti-Thy1 nephritis from days 1 to 5 and established gene expression profiles at each time point using microarrays to explore the development of inflammation.According to the gene expression profiles,macrophage infiltration(triggered by CCL2 activation)was evident on day 1 and enhanced inflammation over the next few days.We screened for genes with expression levels similar to CCL2 and found that the upregulation of the circadian gene albumin D-site-binding protein(DBP)was involved in CCL2 activation in mesangial cells.More importantly,CCL2 expression showed oscillatory changes similar to DBP,and DBP induced peak CCL2 expression at 16:00 a clock on day 1 in the anti-Thy1 nephritis model.We knocked down DBP through transfection with a small interfering RNA(siRNA)and used RNA sequencing to identify the DBP-regulated TNF-α-CCL2 pathway.We performed chromatin immunoprecipitation sequencing(ChIP-Seq)and the dual luciferase assay to show that DBP bound to the TRIM55 promoter,regulating gene expression and in turn controlling the TNF-α-CCL2 pathway.In conclusion,DBP-regulated circadian CCL2 expression by the TRIM55-TNF pathway in injured mesangial cells at an early stage,which promoted macrophage recruitment and in turn triggered infiltration and inflammation in a model of anti-Thy1 nephritis.

Risk factors of prognosis after acute kidney injury in hospitalized patients

The risk factors, especially laboratory indicators, of prognosis after acute kidney injury （AK1） remain unclear. We conducted a retrospective survey of Chinese People＇s Liberation Army General Hospital from January 1, 2012 to December 31, 2012 according to the AKI diagnosis standard issued by Kidney Disease Improving Global Outcomes. The epidemiological features and factors influencing hospital mortality and renal function recovery were evaluated through logistic regression analysis. Among 77 662 cases of hospitalized patients, 1387 suffered from AKI. The incidence rate and mortality of AKI were 1.79~ and 14.56%, respectively. Multivariate logistic regression analysis revealed that high AKI stage, age greater than 80 years, neoplastic disease, low cardiac output, increased white blood cell count, and decreased platelet count and serum albumin levels were the risk factors affecting the mortality of AKI patients. Conversely, body mass index between 28 and 34.9 was a protective factor. Increased AKI stage, tumor disease, post-cardiopulmonary resuscitation, and RRT were the risk factors of renal function recovery upon discharge. In addition to traditional risk factors, white blood cell count, platelet count, albumin, and BMI were the predictors of the mortality of AKI patients. No laboratory indicators were found to be the risk factors of renal function recovery in AKI patients.

Comparative analysis of membranous and other nephropathy subtypes and establishment of a diagnostic model

This study aimed to compare clinical features between membranous nephropathy (MN) and nonmembranous nephropathy (non-MN), to explore the clinically difTerential diagnosis of these two types, and to establish a diagnostic model of MN. After renal biopsy was obtained, 798 patients were divided into two groups based on their examination results: primary MN group (n = 248) and non-MN group (n = 550). Their data were statistically analyzed. Logistic regression analysis indicated that anti-PLA2R antibodies, IgG, and Cr were independently correlated with MN, and these three parameters were then used to establish the MN diagnostic model. A receiver operating characteristic (ROC) curve confirmed that our diagnostic model could distinguish between patients with and without MN, and their corresponding sensitivity, specificity, and AUC were 79.9%, 89.4%, and 0.917, respectively. The cutoff value for this combination in MN diagnosis was 0.34. The established diagnostic model that combined multiple factors shows a potential for broad clinical applications in differentiating primary MN from other kidney diseases and provides reliable evidence supporting the feasibility of noninvasive diagnosis of kidney diseases.

Single-cell RNA-Seq analysis identified kidney progenitor cells from human urine

Dear Editor,Urine passes through the entire kidney and urinary tract system starting from the glomerulus and ending to the urethra.Cells in the kidney and urinary tract could be exfoliated from the epithelium into the urine,while leukocyte could infiltrate from the local tissue into the urine,which makes the urine a useful subject for clinical evaluation of relevant diseases.

Immune Responses to Six Synthetic Peptides of Capsid Protein with Sera from HIV-1 Infected Individuals

Many B cell epitopes within p24 of human immunodeficiency virus type 1 （HIV-1） were identified, while most of them were determined by using murine monoclonal antibodies reacting with overlapping peptides of p24. Therefore these epitopes may not represent the actual epitopes recognized by the HIV-1 infected individuals. In the present study, immune responses of 67 HIV-1 positive sera from Yunnan Province, China to five peptides on p24 of HIV-1 and one of HIV-2 were analyzed. All of 67 sera did not recognize peptide GA-12 on HIV-1 and peptide AG-23 on HIV-2, which indicated that GA-12 was not human B cell epitope and AG-23 did not cross-react with HIV-1 positive serum. Except 13 sera （19.4%）, all remaining sera did not recognize peptides NI-15, DR-16, DC-22 and PS-18, which indicated that these four peptides represented B cell linear epitopes of HIV-1 p24 in some HIV-1 infected individuals but not the immuno-dominant epitopes in most individuals.

Hypertension in patients with CKD in China： clinical characteristics and management

Chronic kidney disease （CKD） is a life-threatening disease that results in end-stage renal disease （ESRD） and consumes substantial health resources. In China, the prevalence of CKD in the general population is 10.8% and increases gradually with age [1]. CKD also causes hypertension.

Sugemule-10:Source of prescription and modern clinical application

Sugemule-10,one of the traditional Mongolian medicine(TMM)formulae,is derived from Four Medical Classics(Vol.4)and composed of 10 Mongolian medicines.It is used to treat kidney cold,low back pain,urinary obstruction,kidney/bladder stones,and is the main prescription for kidney cold.The current research on Sugemule-10 is mostly focused on its clinical efficacy,and few papers are available upon its historical changes.Therefore,we systematically reviewed Sugemule-10 from the aspects of prescription source,prescription interpretation,efficacy evolution,and modern clinical applications.

Cell Cycle Arrest Protein CDKN2C Is Not an HBV Host Factor

Dear Editor,Hepatitis B virus(HBV)infection remains a major cause of liver diseases worldwide,which affects approximately 250 million people globally(Yuen et al.2018).Individuals with chronic HBV infection are at high risk of developing liver cirrhosis,and ultimately hepatocellular carcinoma(HCC).HBV is a Hepadnavirus DNA virus,which specifically infects and efficiently replicates in hepatocytes.It has been proposed for decades that HBV replicates preferentially in non-dividing quiescent hepatocytes(Ozer et al.1996).