Guided by Academician Zhang Boli's theory of "dampness， turbidity， phlegm， and fluid-related diseases"，this paper elaborated on the pathogenesis and syndrome differentiation treatment of heart failure from the perspective of the "spleen-mitochondria". It analyzed the essential similarities between "spleen-mitochondria" and "dampness， turbidity， phlegm， and fluid-related diseases"， as well as their close association with the onset of heart failure. Furthermore，it explored the connection between spleen function and mitochondrial function in traditional Chinese medicine （TCM），positing that the spleen's role in transportation and transformation is analogous to mitochondrial material metabolism and energy conversion，with spleen deficiency closely related to mitochondrial dysfunction. It thus concluded that mitochondrial material metabolism and energy conversion represent the microscopic essence of the spleen's role in transportation and transformation，and mitochondrial dysfunction is a contributing factor to pathological products like dampness and turbid phlegm，which are closely associated with the occurrence of heart failure. The four elements of dampness，turbidity，phlegm，and fluid are a series of related symptoms resulting from abnormal fluid transportation and transformation，serving as both factors in the onset of heart failure and the core pathological basis for its deterioration. Therefore，during the treatment of heart failure，it is essential to regulate mitochondrial function. Early intervention should focus on eliminating dampness and turbidity to improve mitochondrial function and restore normal energy metabolism. In the middle and late stages，emphasis should be placed on resolving phlegm，promoting blood circulation，warming Yang，and reducing water retention to alleviate mitochondrial damage and improve cardiac function. Supporting Qi and strengthening the spleen should be a continuous approach，and treatment should be adjusted to enhance mitochondrial function and stabilize the condition，thereby improving prognosis. This paper discussed the role of the spleen and mitochondria in the pathogenesis of heart failure，examined the evolution of heart failure mechanisms from the perspective of dampness， turbidity， phlegm， and fluid-related diseases，and proposed a phased treatment strategy. It enriched the theory of dampness， turbidity， phlegm， and fluid-related diseases and offered new strategies for heart failure treatment. However，in practical application，TCM strategies for treating heart failure need to be integrated with modern medical approaches to provide a more solid scientific foundation for treatment.

Objective To explore the diagnosis of clinically suspicious von Willebrand disease(vWD)in a family and its pathogene-sis.Methods The pedigree information and the biological specimen were collected from the clinically suspected VWD patient and her family members(4 persons in total)in Peking University First Hospital.The levels of platelet count(PLT),activated partial thrombo-plastin time(APTT),vWF antigen(vWF:Ag),vWF activity(vWF:Ac)and FⅧ activity(FⅧ:C)were detected,and vWF risto-cetin cofactor(vWF:RCo)assay,ristocetin-induced platelet aggregation assay(RIPA)and vWF collagen binding(vWF:CB)assay were performed for phenotype diagnosis.The peripheral blood genomic DNAs were extracted from the proband and her family members to perform whole-exome sequencing for identifying the mutation of vWF gene,The mutation site was analyzed by using bioinformation tools to explore the pathogenesis of the proband.Results The APTT of proband(m 1)was slightly prolonged and her vWF:Ag,vWF:Ac,vWF:RCo and vWF:CB were significantly decreased.There was no obvious aggregation in RIPA assay(1.0 mg/mL and 1.25 mg/mL).In her father(Ⅱ3),APTT,FⅧ:C,vWF:Ag,vWF:Ac and vWF:CB were normal,but vWF:RCo was slightly decreased.In her mother(Ⅱ4),APTT,FⅧ:C,vWF:Ag,vWF:RCo and vWF:CB were all normal,but vWF:Ac significantly decreased.In her brother(Ⅲ2),APTT and FⅧ:C were normal,but vWF:Ag,vWF:Ac,vWF:RCo and vWF:CB were reduced to varying degrees.In all the family members(father,mother and brpther),no apparent aggregation in RIPA(1.0 mg/mL)was shown.Genetic analysis showed that the proband(Ⅲ1)carried a compound heterozygous mutation of vWF gene c.7288-9T＞G and c.1117C＞T,her father(Ⅱ3)carried vWF gene c.7288-9T＞G heterozygous mutation,and vWF gene c.1117C＞T heterozygous mutation was presented in both mother(Ⅱ4)and brother(Ⅲ2).Conclusion According to the results of laboratory tests,the proband was diagnosed as type 2A vWD.The hetero-zygous mutation in vWF gene c.1117C＞T and c.7288-9T＞G may be the molecular mechanism leading to type 2A vWD in the proband.

Objective:To investigate the effect of tripartite motif-containing protein 59 (TRIM59) on glucose metabolism in macrophages and its role in regulating hypoxia-inducible factor-1α (HIF-1α)/IL-10 axis in macrophages under inflammatory conditions.Methods:The differentially expressed genes between macrophages with high expression of TRIM59 and control cells transfected with empty TRIM59 plasmid were analyzed by GO and KEGG. The expression of HIF-1α by RAW264.7 macrophages with high expression of TRIM59 was detected at different time points after lipopolysaccharide (LPS) stimulation by RT-qPCR and Western blot. Bone marrow was isolated from TRIM59-cKO and TRIM59 flox/flox mice and induced to differentiate into bone marrow-derived macrophages (BMDMs). These BMDMs were stimulated with LPS and the supernatants of cell culture were collected at 3, 6, 12 and 24 h after stimulation to detect IL-10 level by ELISA. In addition, mouse models of cecal ligation and puncture (CLP) were established, and bronchoalveolar lavage fluid (BALF) samples were collected at the same time points to detect IL-10 level by ELISA. Histopathological changes in lung tissues were observed after HE staining. Results:There was a significant change in glucose metabolism-related genes in macrophages with high expression of TRIM59, and the content of lactic acid increased significantly. Compared with the control group, the expression of HIF-1α at mRNA level in BMDMs from TRIM59-cKO mice decreased after LPS stimulation ( P<0.05); the level of IL-10 increased at 3 h and 24 h in the TRIM59-cKO group, but there was no significant difference in IL-10 level at 6 h or 12 h between the two groups. In the TRIM59-cKO mouse model of CLP, the levels of IL-10 in the BALF samples increased with time, but decreased at 24 h. The level of IL-10 was higher in the TRIM59-cKO mouse model group than that in the control group at each time point ( P<0.05 or P<0.01). Conclusions:TRIM59 can inhibit inflammation and lung injury by decreasing HIF-1α-mediated lactate secretion and IL-10 expression in macrophages. This study provides a new idea for developing novel anti-sepsis drugs based on TRIM59.

Purpose: The prognosis of patients with hepatocellular carcinoma (HCC) and portal vein tumor thrombus (PVTT) is extremely poor, and systemic therapy is currently the mainstream treatment. This study aimed to assess the efficacy and safety of lenvatinib combined with anti–programmed cell death-1 antibodies and transcatheter arterial chemoembolization (triple therapy) in patients with HCC and PVTT. Materials and Methods: This retrospective multicenter study included patients with HCC and PVTT who received triple therapy, were aged between 18 and 75 years, classified as Child-Pugh class A or B, and had at least one measurable lesion. The overall survival (OS), progression-free survival (PFS), objective response rates, and disease control rates were analyzed to assess efficacy. Treatment-related adverse events were analyzed to assess safety profiles. Results: During a median follow-up of 11.23 months (range, 3.07 to 34.37 months), the median OS was greater than 24 months, and median PFS was 12.53 months. The 2-year OS rate was 54.9%. The objective response rate and disease control rate were 69.8% (74/106) and 84.0% (89/106), respectively; 20.8% (22/106) of the patients experienced grade 3/4 treatment-related adverse events and no treatment-related deaths occurred. The conversion rate to liver resection was 31.1% (33/106), with manageable postoperative complications. The median OS was not reached in the surgery group, but was 19.08 months in the non-surgery group. The median PFS in the surgery and non-surgery groups were 20.50 and 9.00 months, respectively. Conclusion Triple therapy showed promising survival benefits and high response rates in patients with HCC and PVTT, with manageable adverse effects.

Objective:To explore the mechanism of Ginseng Radix et Rhizoma- Notoginseng Radix et Rhizoma- Chuanxiong Rhizoma medicinal pair in delaying heart aging based on animal experiments, network pharmacology and molecular docking. Methods:Mice were divided into control group, aging group, metformin group and TCM group according to random number table method. All the groups were injected subcutaneously by D-galactose except the control group to build the subacute aging model. Two weeks later, the metformin group was given metformin suspension (150 mg/kg), the TCM group was given Ginseng Radix et Rhizoma- Notoginseng Radix et Rhizoma- Chuanxiong Rhizoma lyophilized powder solution (650 mg/kg), and the control group and aging group were given an equivalent volume of ultrapure water by gastric gavage, once a day, six times a week, for 10 weeks. The level of heart TERT mRNA was detected by PCR; the expression of heart p53 was observed by immunohistochemical staining; the morphology of heart tissue was observed by HE staining. TCMSP and SwissTargetPrediciton databases were used to retrieve the active components and targets of Ginseng Radix et Rhizoma- Notoginseng Radix et Rhizoma- Chuanxiong Rhizoma medicinal pair; TTD, OMIM, Gene, HAGR, DisGeNET and other data platforms were used to screen the targets of heart aging; after the drug and disease targets were intersected, the active components of them were collected; STRING database, Cytoscape 3.8.0 software, etc. were used to make PPI of the intersection targets, and screen out the key targets; FunRich was used to perform enrichment analysis of cellular components, molecular functions, biological processes, and biological signal pathways for key targets; Schr?dinger Maestro software was used to do the molecular docking of the screened active components and key targets, and docking results were visualized via PyMOL 2.1 software. Results:Experiment results showed that Ginseng Radix et Rhizoma- Notoginseng Radix et Rhizoma- Chuanxiong Rhizoma could significantly ameliorate the damage of aging heart tissues, elevate TERT mRNA level, while significantly reducing the positive expression of p53. A total of 32 active components from the medicinal pair were screened, corresponding to 637 target genes. There were 263 targets for heart aging, and 67 intersection targets of drug active component targets and heart aging targets. 31 key targets were obtained after screening. Enrichment analysis showed that molecular functions were related to transcription factor activity and protein-tyrosine kinase activity. Biological processes involved signal transduction and cell communication. Signaling pathways mainly involved PDGFR-beta, PI3K-Akt, S1P1, Glypican, TRAIL, and Glypican 1. The molecular docking results showed that kaempferol, suchilactone, and ginsenoside Rg5_qt in the medicinal pair had a strong binding ability to p53. Conclusion:Ginseng Radix et Rhizoma- Notoginseng Radix et Rhizoma- Chuanxiong Rhizoma may achieve the effect of delaying heart aging by inhibiting p53 expression, providing a foundation for further research on mechanism of invigorating qi and activating blood circulation drugs to delay heart aging.

ObjectiveTo observe the effects of Cuscutae Semen on the learning and memory ability， N6-methyladenosine （m⁶A）-related modification enzymes and total m⁶A level in hippocampus of the offspring of fear-damaged pregnant rats. MethodForty-five pregnant rats were randomly divided into blank group， model group and Cuscutae Semen group. From the 1^st day to the 19^th day of pregnancy， rats in the model group and the Cuscutae Semen group were induced by observing electric shock of other rats. The Cuscutae Semen group was treated with 5 g·kg^-1·d^-1 Cuscutae Semen decoction （ig）， while the other two groups were treated with the same amount of purified water. The offspring were assigned following the grouping method of their maternal generation. The behavioral changes of the offspring were tested by Morris water maze on 21^st day after birth， and the development of hippocampal neurons was observed by transmission electron microscopy. The mRNA and protein expression levels of methyltransferase-like 3 （METTL3）， METTL14， Wilms tumor 1 associated protein （WTAP）， fat mass and obesity-associated protein （FTO） and Alk B homologue 5 （ALKBH5） were detected by Real-time polymerase chain reaction （Real-time PCR）， Western blot and immunohistochemistry （IHC）. The total content of m⁶A in hippocampus was determined by high performance liquid chromatography tandem mass spectrometry （LC-MS/MS）. ResultCompared with the conditions in the blank group， the average latency duration in the model group was prolonged， and the number of entries in the target quadrant， the target quadrant duration and the number of crossing the platform were decreased （P<0.01）. Additionally， the model group had seriously damaged structure of hippocampal CA1 and CA3 neurons， swollen mitochondria， expanded endoplasmic reticulum， and small number of synapses with some having blurred structure， and the expression levels of METTL3， METTL14， FTO， ALKBH5 as well as the total m⁶A level were lower than those in the blank group （P<0.05，P<0.01）. Compared with the model group， the Cuscutae Semen group had shortened average latency duration， increased number of entries in the target quadrant， target quadrant duration and number of crossing the platform （P<0.01）， alleviated damage of hippocampal CA1 and CA3 neurons， fine structure of mitochondrial and endoplasmic reticulum， and clear， intact and dense synapses. And the expression levels of METTL3， METTL14， FTO as well as the total level of m⁶A were up regulated， while the expression level of ALKBH5 was down regulated in the Cuscutae Semen group （P<0.05，P<0.01）. ConclusionCuscutae Semen improved the learning and memory ability of the offspring of the rats affected by fear damaging kidney during pregnancy， protected hippocampal neurons， and up-regulated the expression levels of METTL3， METTL14， FTO and the total m⁶A level in hippocampus.

Objective This study examined the research status and development process of FU Qingzhu’s Obstetrics and Gynecology (Fu Qing Zhu Nv Ke,《傅青主女科》, FQZNK) in the past 40 years with bibliometrics and visual analysis. Methods Retrieved all related literature in the research field of FQZNK from the domestic and foreign databases: China National Knowledge Infrastructure (CNKI), China Science and Technology Journal Database (VIP), Wanfang Database, and Web of Science (WOS) core database, including Science Citation Index Expanded (SCIE), Social Sciences Citation Index (SSCI), and Arts & Humanities Citation Index (A&HCI). The search range was from January 1, 1980 to March 10, 2021. In addition, bibliometrics and CiteSpace 5.7.R2 software were used to analyze literature types, published journals, cited literature, the number of author publications, co-author networks, co-institution networks, keyword co-occurrence networks, keyword clusters, and keyword bursts. Results A total of 678 valid records were included in the final dataset. Literature types, high publication journals, highly cited literature, high-yield institutions, high-yield research teams, and high-productivity scholars in this research field were found through bibliometrics. Literature types can be divided into four categories, among which 451 are theoretical studies on academic thoughts of FQZNK, accounting for 66.5% of the included journals. The Journal of Shanxi Traditional Chinese Medicine had the largest volume of published articles (61), accounting for 9.0% of the total number of the included journals. The most cited literature was ZHOU Mingxin’s article “Using the quantitative method to discuss author’s authenticity and formula characteristics of FU Qingzhu’s Obstetrics and Gynecology”, which was cited 94 times. Hunan University of Chinese Medicine, the institution with the most publications, published 45 articles, and YOU Zhaoling, the most published author, published 33 articles. Moreover, it was found that most high-yield researchers came from high-yield institutions and that Hunan University of Chinese Medicine had the most research on FQZNK. Keyword co-occurrence analysis revealed that the keyword “FQZNK” had the highest frequency (597 times) and the highest centrality (1.00). Keyword cluster analysis used the Log-Likelihood Ratio (LLR) algorithm to form eleven important clusters: #0 treatment aiming at its root causes, #1 gynecopathy, #2 Siwu Decoction (四物汤), #3 FU Qingzhu, #4 post-partum, #5 infertility, #6 dysmenorrhea, #7 sterility, #8 coordinate the heart and kidney, #9 Danggui Buxue Decoction (当归补血汤), and #10 treatment. It was found that the prescriptions of FQZNK were studied mainly before 2000, the theoretical studies were mainly conducted before 2010, and its clinical application was mainly explored from 2010 until now. Diseases such as dysmenorrhea, morbid vaginal discharge, infertility, metrorrhagia, and polycystic ovarian syndrome (PCOS) have recently become popular topics in this field. Conclusion The current study provides more scientific, accurate, and comprehensive scientific support for further research and development of traditional Chinese medicine (TCM) in FQZNK. With this foundation, people can use burst detection to ascertain the current hotspots in research, get their development trends, and forecast future research directions. In addition, infertility, morbid vaginal discharge, flooding, and PCOS treatments based on TCM syndrome differentiation are currently popular research topics for FQZNK.

Objective:To evaluate the accuracy of pRIFLE criterion and KDIGO criterion for the diagnosis of acute kidney injury (AKI) after radical operations for tetralogy of Fallot in children from the perspective of postoperative outcomes.Methods:A total of 375 children, aged<8 yr, undergoing radical operations for tetralogy of Fallot, were selected continuously and retrospectively. According to the pRIFLE and KDIGO diagnostic criteria, postoperative AKI was diagnosed, and the children were classified into different AKI grades. The prognostic parameters (postoperative mechanical ventilation time, duration of intensive care unit (ICU) stay, postoperative length of hospital stay, total costs of hospitalization, and in-hospital mortality, etc.) were collected, and the differences among different AKI grades were compared. Logistic regression method was used to analyze the risk factors for prolonged postoperative length of hospital stay (≥14 days) when two different criteria were used to diagnose AKI. The children diagnosed as non-AKI by KDIGO criterion were further confirmed using pRIFLE criterion, and the prognostic parameters in the children who were diagnosed as AKI and non-AKI were compared.Results:When two different criteria were used to diagnose AKI after radical resection for tetralogy of Fallot, the incidence was 56.8% (pRIFLE criterion) and 40.0% (KDIGO criterion). AKI diagnosed according to the two criteria was the independent risk factor for prolonged postoperative length of hospital stay, and the levels of all the prognostic parameters (postoperative mechanical ventilation time, duration of ICU stay, postoperative length of hospital stay, total costs of hospitalization, and in-hospital mortality) were significantly higher in AKI children than in non-AKI (AKI grade 0) children ( P<0.01). Among the 225 children diagnosed as non-AKI according to the KDIGO criterion, 63 cases were diagnosed as AKI and 162 cases as non-AKI according to the pRIFLE criterion, however, there was no significant difference in each prognostic parameter between children with AKI and non-AKI ( P>0.05). Conclusion:The pRIFLE criterion has a higher sensitivity, while the KDIGO criterion produces better accuracy when used to evaluate the diagnosis of AKI after radical operation for tetralogy of Fallot in children from the perspective of postoperative outcomes.

Exonuclease Ⅷ (Exo Ⅷ), an ATP-independent dsDNA 5'-3' exonuclease, is a candidate protein with great application value for in vitro DNA recombination. However, the application of Exo Ⅷ in DNA recombination in vitro has not been reported. In this study, the recombinant expression vector of the truncated Exo Ⅷ (tExo Ⅷ) with the full exonuclease activity was built and used to achieve the overexpression of tExo Ⅷ in Escherichia coli. Based on the purified tExo Ⅷ protein with high-purity, the feasibility of tExo Ⅷ applied in vitro DNA recombination and effects of the reaction temperatures, reaction duration, and homology arm lengths were examined. The results showed that tExo Ⅷ was highly expressed in soluble form in E. coli. One liter of bacterial culture yielded 92.40 mg of purified tExo Ⅷ with the specific activity of 1.21×10⁵ U/mg. In a 10 μL recombination system containing 2.5 U tExo Ⅷ, the highest cloning efficiency was achieved in a reaction at 25 °C for 12.5 min and followed by incubation at 50 °C for 50 min. With addition of Pfu DNA polymerase, the homology arm extension strategy can effectively improve the recombination efficiency. Using competent E. coli Mach1 T1 with 2.2×10⁶ cfu/μg transformation efficiency as recipient cell, the recombination of a 1 kb fragment with a 21 bp homology arm and a 5.8 kb linearized vector can form about 1.1×10⁴ recombinant clones per μg vector, and the positive rates was over 80%. The recombination efficiency was increased with the increasing length of homology arm ranged from 8 to 21 bp. Under the optimal reaction condition, only 8 bp homology arm can still achieve valid DNA recombination. This novel in vitro DNA recombination system mediated by tExo Ⅷ was particularly characterized by its easy preparation, no limitation on restriction sites and high recombination cloning efficiency. All results revealed that the new efficient gene cloning system has potential application in the field of molecular biology.
Cloning, Molecular ; Escherichia coli ; genetics ; Exonucleases ; genetics ; Recombinant Proteins ; metabolism ; Recombination, Genetic

Objective: To analyze the genetic characterization of 2B genotype rubella virus strains first isolated in Liaoning province. Methods: Vero/Slam cells were used to isolate rubella viruses from throat swabs. Fragments of the E1 gene (739 nucleotides) were amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the PCR products were sequenced and analyzed. The phylogenetic trees were constructed with rubella 2B genotypes reference strains. Results: RVi/Liaoning.CHN/13.18/1[2B] was isolateded from the case which had a history of international travel and probably imported from abroad. RVi/Liaoning.CHN/13.18/1[2B] was in the same 1ineage with RVs/Edinburgh.GBR/6.12[2B] and RVs/Kawasaki.JPN/17.11/1[2B], but far from the epidemic strains isolated in China in recent years. RVi/Liaoning.CHN/14.18/1[2B] belonged to the same lineage with rubella virus 2B genotype strains isolated in China from 2014 to 2015, and had the highest homology with RVi/Chouzhou.Anhui.CHN/17.15/2[2B] and RVi/Shaoyang.Zhejiang. CHN/13.13[2B]. Conclusions It was the first time to isolate the 2B genotype rubella virus in Liaoning province and the strain belonged to different 1ineage.