Bibenzyls, a kind of important plant polyphenols, have attracted growing attention for their broad and remarkable pharmacological activities. However, due to the low abundance in nature, uncontrollable and environmentally unfriendly chemical synthesis processes, these compounds are not readily accessible. Herein, one high-yield bibenzyl backbone-producing Escherichia coli strain was constructed by using a highly active and substrate-promiscuous bibenzyl synthase identified from Dendrobium officinale in combination with starter and extender biosynthetic enzymes. Three types of efficiently post-modifying modular strains were engineered by employing methyltransferases, prenyltransferase, and glycosyltransferase with high activity and substrate tolerance together with their corresponding donor biosynthetic modules. Structurally different bibenzyl derivatives were tandemly and/or divergently synthesized by co-culture engineering in various combination modes. Especially, a prenylated bibenzyl derivative ( 12) was found to be an antioxidant that exhibited potent neuroprotective activity in the cellular and rat models of ischemia stroke. RNA-seq, quantitative RT-PCR, and Western-blot analysis demonstrated that 12 could up-regulate the expression level of an apoptosis-inducing factor, mitochondria associated 3 (Aifm3), suggesting that Aifm3 might be a new target in ischemic stroke therapy. This study provides a flexible plug-and-play strategy for the easy-to-implement synthesis of structurally diverse bibenzyls through a modular co-culture engineering pipeline for drug discovery.

Objective:To explore the molecular mechanism of the effect of the histone methylase zeste gene enhancer homolog 2 (EZH2) on the proliferation and apoptosis of human hypertrophic cardiomyocytes AC16.Methods:The AC16 hypertrophic cardiomyocyte model was constructed by adding angiotensin Ⅱ to the AC16 cell culture medium. The cells were divided into four groups, including the blank control group, the angiotensin Ⅱ group, the empty vector + angiotensin Ⅱ group, and the EZH2 overexpression + angiotensin Ⅱ group. The expression levels of EZH2 and brain natriuretic peptide ( BNP) genes were measured using fluorescent quantitative PCR. The EZH2, trimethylation of lysine at position 27 of histone H3 (H3K27me3), and BNP proteins expression were detected by Western Blot. The MTS method was used to detect the proliferation of AC16 cell. The Annexin V-FITC/PI double staining method was used to detect the apoptosis of AC16 cell. Results:Compared with the blank control group, the expression levels of EZH2 and H3K27me3 in the angiotensin Ⅱ group were decreased, the expression level of BNP was increased, cell proliferation was decreased, and apoptosis was increased (all P < 0.001). Compared with the empty vector + angiotensin Ⅱ group, the expression levels of EZH2 and H3K27me3 in the EZH2 overexpression + angiotensin Ⅱ group were increased, the expression level of BNP was decreased, the cell proliferation level was increased, and the apoptosis level was decreased (all P < 0.001). There was no significant difference between the angiotensin Ⅱ group and the empty vector + angiotensin Ⅱ group (all P > 0.05). Conclusions:Histone methylase EZH2 has an effect on the proliferation and apoptosis of AC16 cell, providing a reference for the treatment of myocardial hypertrophy and revealing the exact pathogenesis of myocardial hypertrophy.

Orthodontically induced tooth root resorption (OIRR) is a serious complication during orthodontic treatment. Stimulating cementum repair is the fundamental approach for the treatment of OIRR. Parathyroid hormone (PTH) might be a potential therapeutic agent for OIRR, but its effects still lack direct evidence, and the underlying mechanisms remain unclear. This study aims to explore the potential involvement of long noncoding RNAs (lncRNAs) in mediating the anabolic effects of intermittent PTH and contributing to cementum repair, as identifying lncRNA-disease associations can provide valuable insights for disease diagnosis and treatment. Here, we showed that intermittent PTH regulates cell proliferation and mineralization in immortalized murine cementoblast OCCM-30 via the regulation of the Wnt pathway. In vivo, daily administration of PTH is sufficient to accelerate root regeneration by locally inhibiting Wnt/β-catenin signaling. Through RNA microarray analysis, lncRNA LITTIP (LGR6 intergenic transcript under intermittent PTH) is identified as a key regulator of cementogenesis under intermittent PTH. Chromatin isolation by RNA purification (ChIRP) and RNA immunoprecipitation (RIP) assays revealed that LITTIP binds to mRNA of leucine-rich repeat-containing G-protein coupled receptor 6 (LGR6) and heterogeneous nuclear ribonucleoprotein K (HnRNPK) protein. Further co-transfection experiments confirmed that LITTIP plays a structural role in the formation of the LITTIP/Lgr6/HnRNPK complex. Moreover, LITTIP is able to promote the expression of LGR6 via the RNA-binding protein HnRNPK. Collectively, our results indicate that the intermittent PTH administration accelerates root regeneration via inhibiting Wnt pathway. The lncRNA LITTIP is identified to negatively regulate cementogenesis, which activates Wnt/β-catenin signaling via high expression of LGR6 promoted by HnRNPK.
Mice ; Animals ; Cementogenesis ; Wnt Signaling Pathway ; beta Catenin/metabolism* ; Heterogeneous-Nuclear Ribonucleoprotein K/metabolism* ; RNA, Long Noncoding/genetics* ; Parathyroid Hormone ; Receptors, G-Protein-Coupled/metabolism*

Objective:To investigate the clinical effect of vascular pressurisation and super reflux on free anterolateral thigh flap (ALTF).Methods:From January 2017 to September 2021, the Department of Hand and Foot Microsurgery, the Second Affiliated Hospital of Hainan Medical College had treated 31 cases of soft tissue defects of the limbs. The patients were 23 males and 8 females, aged 4-76 years old at 40 years old in average. All the patients received transfer of free ALTFs carrying 2 groups of blood vessels of the descending and oblique branches of lateral circumflex femoral artery (LCFA). The vascular pressurisation and super-reflux techniques were applied in wound repair. Soft tissue defect area 7 cm × 5 cm-22 cm × 10 cm. The sizes of flaps were 8 cm×6 cm-23 cm×11 cm. All the donor sites were sutured directly. Fifteen patients had inner flap pressurisation (or super-reflux) and the rest of 16 patients had external flap pressurisation (or super-reflux). After surgery, scheduled follow-ups were conducted through outpatient clinic, telephone and WeChat reviews or home visits to evaluate the efficacy of wound repair.Results:All the 31 flaps survived, except 2 flaps that had mild infection after surgery. The wounds of donor and recipient sites healed completely. The time of follow-up was 3-55 months. The skin of flaps achieved good texture, colour, lustre and appearance. TPD of the flaps ranged 7-12 mm.Conclusion:Vascular pressurisation and super reflux technique are stable and reliable in the clinical application of free ALTF to repair soft tissue defects of limbs.

Cold atmospheric plasma (CAP), a redox modulation tool, is capable of inhibiting a wide spectrum of cancers and has thus been proposed as an emerging onco-therapy. However, with incremental successes consecutively reported on the anticancer efficacy of CAP, no consensus has been made on the types of tumours sensitive to CAP due to the different intrinsic characteristics of the cells and the heterogeneous design of CAP devices and their parameter configurations. These factors have substantially hindered the clinical use of CAP as an oncotherapy. It is thus imperative to clarify the tumour types responsive to CAP, the experimental models available for CAP-associated investigations, CAP administration strategies and the mechanisms by which CAP exerts its anticancer effects with the aim of identifying important yet less studied areas to accelerate the process of translating CAP into clinical use and fostering the field of plasma oncology.

【Objective】 To understand the distribution and gene frequency of main red blood cell blood groups in Lahu ethnic minority and analyze the genetic characteristics of Lahu people. 【Methods】 1) ABO forward and reverse typing had been performed by microplate method; 2) Rh, MN, H, P1Pk and Mur antigen were tested by the tube method. If the ABO forward and reverse typing were incompatible, the tube method was used for confirmation. 【Results】 The distribution characteristics of blood group and gene frequency in Lahu ethnic minority were as follows: B>O>A>AB for ABO, with genotype frequency as p 11.1%, q 27.5% and r 61.4%; the frequency of Rh genotype was CDe 83.3%, cDE 12.0%, cDe 2.42%, CDE 2.32%, CdE 0%, Cde 0%, cdE 0% and cde 0%; M > MN>N for MN blood group, with genotype frequency as M 75.26% and N 24.74%; P1

Objective:To explore the effects of pre- and post-diagnosis of malignant neoplasms total cholesterol difference on all-cause death in the Kailuan study population.Methods:A prospective cohort study method was used to observe on the job and retired workers who were diagnosed with malignant neoplasms during the healthy physical examination and had the complete data of total cholesterol pre- and post-diagnosis of malignant neoplasms at Kailuan (Group) Limited Liability Corporation from June 2006 to December 2015. Medical insurance system and medical records case management system of Kailuan General Hospital were used to retrieve the confirmed diagnosis time of participants with malignant neoplasms. SAS 9.4 software was used to extract the physical examination data of participants diagnosed as malignant neoplasms before and after 2 years. The whole participants were divided into 4 groups according to the quartile of the total cholesterol difference pre- and post-diagnosis of cancer. The first quartile group: the total cholesterol difference <-0.93 mmol/L (823 cases), the second quartile group: the total cholesterol difference≥-0.93 mmol/L and <-0.21 mmol/L (811 cases), the third quartile group: the total cholesterol difference≥-0.21 mmol/L and <0.49 mmol/L (832 cases), the forth quartile group: the total cholesterol difference≥0.49 mmol/L (833 cases). The incidence density was used to calculate the mortality of different total cholesterol difference quartile groups, and the Cox proportional hazards model was used to analyze the influencing factors of all-cause death of all quartile groups.Results:Totally, 1 564 cases had all-cause death during median 2.46 years of follow-up time. There were 481, 440, 333, 310 death cases respectively of 4 quartile groups. The death rates of all quartile groups were 230.10/1 000 person-years, 163.90/1 000 person-years, 115.34/1 000 person-years, and 83.44/1 000 person-years respectively ( χ2 = 604.62, P < 0.001). After adjusting for confounding factors, the Cox proportional hazards model analysis showed that compared with the first quartile group, the hazard ratio with the 95% confidence interval of all-cause death in the second, third, and fourth quartile groups was 0.86 (0.76-0.98), 0.62(0.54-0.72) and 0.58 (0.50-0.67) respectively (all Ptrend < 0.001). After adjusting for confounding factors of patients with cancer in different location, the forth quartile group except for cancer in thyroid, breast, prostate, testis and urinary system, all Ptrend was less than 0.05 compared with that of the first quartile group. Conclusion:Among Kailuan study population, the smaller decline or even rise in total cholesterol level of post-diagnosis compared with that of pre-diagnosis is a protective factor of all-cause death.

Objective:To study the HPLC fingerprints of Coptidis Rhizoma- Magnoliae Officmalis Cortex formula granules and the differences of active ingredients in different proportions; To explore the content changes of key components in different proportions of Coptidis Rhizoma- Magnoliae Officmalis Cortex. Methods:HPLC was used to determine the contents of several alkaloids and total phenol of Magnolia officinalis in Coptidis Rhizoma- Magnoliae Officmalis Cortex formula granules and their fingerprints, and the similarity evaluation, cluster analysis and principal component analysis were performed. Results:The similarity of fingerprint of 10 batches of Coptidis Rhizoma- Magnoliae Officmalis Cortex was > 0.950. 17 common peaks were identified, and 6 components were identified. Compared with single medicine, the contents of alkaloids and total phenols in the Coptidis Rhizoma- Magnoliae Officmalis Cortex formula granules were significantly reduced. The contents of multiple alkaloids and total phenols in the Coptidis Rhizoma- Magnoliae Officmalis Cortex formula granules in different proportions were different, and the contents of alkaloids and total phenols were the highest when the proportion of Coptidis Rhizoma- Magnoliae Officmalis Cortex was 2∶1. Conclusion:The contents of main components of Coptidis Rhizoma- Magnoliae Officmalis Cortex formula granules with different proportions are different, which can provide a certain basis for studying the compatibility mechanism of TCM couplet medicines.

【Objective】 To develop a novel screening reagent for -D- phenotype preliminary screening based on the difference in RhD antigen expression level of -D- phenotype and normal RhD phenotype. 【Methods】 RhD antigen expression of -D-phenotype and Rh D-- gene carrier were detected by flow cytometry. By adjusting the concentration of polybrene in the screening system, the red blood cells with high RhD antigen expression level agglutinated, and the preliminary screening of the -D-phenotype and its gene carriers was realized. 【Results】 According to the quantitative results of immunofluorescence intensity (MFI) analysis by flow cytometry, the expression level of RhD antigen in -D- phenotype cells (284 360±16 698, n=3) was about 3 times normal RhD positive cells (98 642±35 908, n=9)(P<0.01), while RhD antigen expression level of RhD-- gene carrier (181 109±39 455, n=4) was about 2 times normal RhD positive cells(P<0.01). RhD antigen expression (144 538±227 445, n=7) of the positive cells screened by 15 μL 3% fresh red blood cell suspension and screening system 35 μL (1 μL IgG anti-D, 29 μL polybrene polybrene, and 5 μL low ionic strength solution) was about 1.5 times normal RhD positive cells. 【Conclusion】 The polybrene preliminary screening system, which can be used for high-throughput screening of -D- phenotype, is a reliable technical method for frequency study of this phenotype.

Objective:To explore the effect of systolic blood pressure (SBP) trajectories on cancers.Methods:The relevant data of 54, 888 employees of Kailuan (Group) Limited Liability Company who participated in the 3 health examinations from 2006-2007, 2008-2009, 2010-2011 were collected and the new onset cancer cases were recorded. The systolic blood pressure trajectory grouping was carried out using the blood pressure measurement values of the 3 physical examinations. The life table method was used to calculate the incidence of cancer, and the multivariate Cox proportional hazard regression model was used to analyze the influence factors of cancer.Results:According to the systolic blood pressure trajectory, 54, 888 subjects were divided into 5 groups, including 14, 326 in the low-stable group, 25, 630 in the moderate-stable group, 5, 390 in the moderate-increasing group, 6, 438 in the elevated-lowering group, and 3, 104 in the elevated-stable group. A total of 1, 070 new onset cancer occurred during the follow-up period of (4.95±0.53) years. The incidence of cancer in the low-stable group, moderate-stable group, moderate-increasing group, elevated-lowering group and elevated-stable group were 1.3% (177/14, 326), 2.2% (491/25, 360), 3.1% (147/5, 390), 2.7% (156/6, 438) and 3.8% (99/3, 104), respectively, the difference was statistically significant ( P<0.001). After adjusting for gender, age, smoking, drinking, physical exercise, body mass index (BMI), fasting blood glucose, total cholesterol, antihypertensive drugs, hypoglycemic drugs, and lipid-lowering drugs, multivariate Cox regression analysis showed that the systolic blood pressure trajectory was related to the incidence of cancer. Compared with the low-stable group, the Hazard ratio ( HR) in the moderate-stable group, moderate-increasing group, elevated-lowering group and elevated-stable group were 1.413, 1.731, 1.557 and 1.907, respectively (all P<0.001). Conclusion:High systolic blood pressure trajectories is the risk factor for cancer.