Podophyllotoxin is an aryl naphthalene lignan-like secondary metabolite, which is widely existed in plants of the Podophyllum. It exhibits a broad spectrum of antitumor activity and holds significant clinical value as a medicinal natural product of plant origin. This review discusses podophyllotoxin from three aspects: plant distribution, biosynthesis and pharmacological effects. Distribution of podophyllotoxin in nature: Podophyllotoxin and its analogues are mainly found in 12 genera of three families, which are Berberidaceae, Linaceae and Cupressaceae, respectively. They are mostly distributed in East Asia and North America, especially in China, which is not only the diversity center of the Podophyllum, but also the distribution center. Among all these plants, the genera Dysosma and Sinopodophyllum are the focus of the relevant research. Biosynthesis of podophyllotoxin: In order to take the alleviation of the pressure of podophyllotoxin that is in short supply of natural resources, and to protect its existing resources, the biosynthesis of podophyllotoxin, especially synthetic biology, has gradually become a new direction of current research. Compared with chemical synthesis which has the shortcoming of low synthesis efficiency, high production cost and cumbersome steps, biosynthesis has a variety of advantages. It’s not limited by resources, easy to cultivate on a large scale and has fast production speed, low production cost and simple genetic manipulation, so there is a broader prospect for the exact purpose of biosynthetic development. The biosynthesis of podophyllotoxin is divided into four steps in general, including the synthesis of coniferyl alcohol, the synthesis of matairesinol, the synthesis of deoxypodophyllotoxin and the synthesis of podophyllotoxin. It requires various enzymes catalyzed during the synthesis process, such as pinoresinol-lariciresinol reductase(PLR), secoisolariciresinol dehydrogenase(SDH) and others. At present, by means of synthetic biology, heterologous production of natural drugs in microbial cell factories, the synthesis of the intermediate coniferyl alcohol has been achieved in Escherichia coli with an increased yield of (124.9±5.1)mg·L−1, and the concentration of (-)-pluviatolide was elevated to 137 mg·L−1. In addition, p-coumaric acid in Saccharomyces cerevisiae was obtained to 71.71 mg·L−1, and in a tobacco plant production chassis, highly purified (-)-deoxypodophyllotoxin was increased to 0.71 mg·g−1 (dry weight) after chromatographic separation. Overall, the utilization of biosynthesis can significantly increase the yield of podophyllotoxin and improve the research and production efficiency of anti-tumor drugs greatly, which brings a more effective strategy for the treatment of related diseases. The pharmacological activity of podophyllotoxin: Podophyllotoxin and its analogues have strong cytotoxicity and they exert good inhibitory effects on the proliferation of multifarious tumor cells, viruses and bacteria. So they are widely applied to the clinical treatment of various types of tumors, such as colorectal cancer, lung cancer, cervical cancer, hemangioma, breast cancer, esophageal cancer, etc. The molecular mechanism of its cancer treatment is mainly in the G1, G2/M to block the cell cycle to make drugs work. Meanwhile, podophyllotoxin has relatively significant toxic side effects and drawbacks that can’t be ignored as a drug with superior therapeutic effect on specific diseases, which tends to increase the risk of bone marrow suppression, chromosomal aberrations and so forth. In conclusion, this literature sums up the distribution of podophyll plants in China, making review of the biosynthetic steps, applications, and biological research of podophyllotoxin, and outlines the pharmacological activity of podophyllotoxin. It aims to set the foundation for the reasonable application of podophyllotoxin resources, the efficient synthesis of podophyllotoxin, and the development of Podophyllum drugs which are highly effective and low toxic. At the same time, this paper gives full play to the clinical medicinal value of podophyllotoxin, which has important influence and far-reaching significance in the treatment of malignant diseases, and promotes the modernization of traditional Chinese medicine.

Objective: To investigate the second-hand smoke behavior at home among smokers, so as to provide the reference for developing home tobacco control strategies. Methods: Permanent residents who were smokers and at the ages of 15 years and above were sampled from 10 streets (townships) in Xuhui District, Shanghai Municipality using the multi-stage random sampling and population-size proportional sampling methods in 2022. Demographic information, smoking status, awareness of second-hand smoke hazards and second-hand smoke behavior at home were collected by questionnaire surveys. Factors affecting second-hand smoke behavior at home were identified using a multivariable logistic regression model. Results: A total of 1 024 smokers were surveyed, including 769 males (75.10%) and 255 females (24.90%). The awareness of hazards of second-hand smoke was 33.59%; the awareness rate of second-hand smoke causing lung cancer in adults was the highest at 76.76%, while the awareness rate of second-hand smoke leading to premature birth and low birth weight in newborns was the lowest at 39.45%. There were 459 smokers with second-hand smoke behavior at home, accounting for 44.82%. Multivariable logistic regression analysis showed that occupation (the retired, OR=2.320, 95%CI: 1.276-4.218), frequency of smoking (often, OR=5.722, 95%CI: 3.977-8.231), smoking duration (a year and above, OR=10.089, 95%CI: 5.508-18.480), electronic cigarette use (occasionally, OR=2.994, 95%CI: 1.283-6.986), living with pregnant women or infants (no, OR=2.171, 95%CI: 1.367-3.448), family indoor smoking restrictions (no restriction, OR=13.926, 95%CI: 7.538-25.727) and awareness of second-hand smoke hazards (unknown, OR=1.562, 95%CI: 1.067-2.287) were the influencing factors for second-hand smoke behavior at home. Conclusion There were 44.82% smokers in Xuhui District with second-hand smoke behavior at home, which was influenced by occupation, living situation, smoking status, family indoor smoking restriction and awareness of second-hand smoke hazards.

Objective To observe the value of cardiac CT angiography(CCTA)for left atrial appendage closure(LAAC).Methods Twelve Labradors were enrolled and underwent LAAC.CCTA examinations were performed before and 7,45 and 90 days after LAAC,and the left atrial appendage(LAA)parameters before and post LAAC occluder compression ratio were recorded.Ninety days after LAAC,the dogs were euthanized for gross anatomical observation,micro CT imaging and pathological examination,and the value of CCTA for LAAC was analyzed.Results Based on CCTA before LAAC,LAA opening depth was(18.8±2.6)mm,the minimum diameter was(16.3±2.0)mm and the maximum diameter was(22.6±3.2)mm,so 28 mm diameter occluders were selected for LAAC.Seven,45 and 90 days after LAAC,no displacement of occluder,thrombi nor device leakage was found on CCTA,while no significant difference of compression ratio was found among different time points(P＞0.05),nor between those measured with CCTA and micro CT 90 days after LAAC(P＞0.05).Ninety days after LAAC,gross dissection revealed complete endothelialization of occlude in 11 dogs and incomplete endothelialization in 1 dog.Pathological examination showed that collagen fibers arranged neatly,with endothelial cells covering the surface,and no inflammatory cell infiltration was observed.Conclusion CCTA could be used to observe LAA and surrounding structures before LAAC,hence guiding to select appropriate occluder,evaluate occlusion effect and monitor complications of LAAC in dogs.

Objective To observe the drainage pathways of interstitial fluid(ISF)and substance clearance pattern in rat brain with fluorescence tracing imaging and treacer-based MRI.Methods Thirty-three male SD rats were randomly divided into fluorescence tracing group(F group,n=18)and treacer-based MRI group(MRI group,n=15),then further divided into thalamic,hippocampal and caudate nucleus subgroups,respectively.Evans blue was injected to rats in F group,and cardiac perfusion was performed after injection,then brain tissue was harvested,and frozen sections were made to observe the drainage pathways of IFS in different subgroups.MRI was performed on rats in MRI group before and after injection of gadolinium-diethylenetriamine pentaacetic acid(Gd-DTPA)to observe signal intensity in ROI of brain regions in different subgroups,the signal unit ratio was calculated,and the changing trend was explored.Results ISF in thalamus,hippocampus and caudate nucleus had different dominant drainage pathways,and the time of tracer reached to adjacent brain regions and whole brain in F group were different.In MRI group,within 4 h after injection of Gd-DTPA,there were differences in direction and clearance rate among tracer in thalamus,hippocampus and caudate nucleus,mainly manifesting as the tracer in thalamus and hippocampus drained to the ipsilateral cortex and lateral ventricle,while the tracer in the caudate nucleus diffused to the cortex and midbrain,and there were differences of the peak time of tracer signal among adjacent drainage brain regions.Conclusion Fluorescence and MR dual-mode imaging showed that there were differences in the dominant drainage pathways of IFS and clearance rates of small molecule substances among hypothalamus,hippocampus and caudate nucleus of rats.

OBJECTIVE To explore the effects of posaconazole combined with proton pump inhibitors （PPI） on the blood concentration and the risk of invasive fungal disease （IFD） in patients with malignant hematological disorder. METHODS In accordance with the random number table method， 40 patients with malignant hematological disorders who were admitted to the hematology department of our hospital between December 2020 and December 2021 were chosen and divided into control group （20 cases） and observation group （20 cases）. The control group received Posaconazole oral suspension alone， while the observation group received Posaconazole oral suspension combined with PPI. The incidence of IFD， attainment rate of blood concentration， the time from the start of prophylaxis to IFD onset， the fatality associated with IFD， treatment of infected patients， and blood concentrations of posaconazole on 7th， 14th， 21st， and 28th day after posaconazole application were compared between 2 groups； the occurrence of adverse events during drug administration in the two groups was recorded. RESULTS The study was stopped because 2 patients in the observation group and 9 patients in the control group received hospital departures after taking posaconazole for fewer than 7 days. The incidence of IFD in the observation group was significantly higher than control group， and the attainment rate of blood concentration in the observation group was significantly lower than control group （P＜0.05）. There was no significant difference in the time from the start of prophylaxis to IFD onset， the fatality associated with IFD， treatment of infected patients and the incidence of adverse events （P＞ 0.05）. The blood concentration of posaconazole in the observation group was significantly lower than control group on 7th day of medication （P＜0.05）； there was no significant in blood concentration of posaconazole between 2 groups on the 14th day of medication （P＞0.05）. CONCLUSIONS Posaconazole combined with PPI can reduce the blood concentration of patients with malignant hematological disorders， increase the risk of IFD. Clinical practice should try to avoid the combination of the two or use them under the guidance of therapeutic drug monitoring.

Objective: To study the effect of long non⁃coding RNA (LncRNA) HAGLR on the expression of NODlike receptor pyrin domain⁃associated protein 3 ( NLRP3 ) inflammasome and fracture healing in tibial fracture (TF) mice and to explore the mechanism . Methods : First , HAGLR in osteoblast MC3T3 ⁃E1 was silenced by in vitro . Cell viability was detected by CCK⁃8 assay , cell apoptosis was detected by TUNEL assay , and the expressions of bone alkaline phosphatase (BALP) and osteocalcin were detected by qPCR . Western blot assay was used to detect the expressions of RUNX2 , phosphorylated RUNX2 ( p ⁃RUNX2 ) , NLRP3 , cysteine aspartic protease 1 (Caspase1) , apoptosis⁃associated spot⁃like protein ( ASC) and interleukin⁃1β . TF mouse models were established by tibial fracture operation in mice . HAGLR was overexpressed in the model mice , and RUNX2 was silenced or an inflammatory body inhibitor MCC950 was added on the basis of overexpression of HAGLR . The expressions of HAGLR and RUNX2 were detected by qPCR , and the expressions of insulin⁃like growth factor (IGF⁃1) were detected by Western blot . microCT was used to measure the volume of mouse callus (MBV) and the total tibial wet weight . Results : The apoptosis rate of MC3T3 ⁃E1 cells increased and the expression levels of RUNX2 ,p⁃RUNX2 , BALP and osteocalcin decreased ( P < 0. 05 ) . The expressions of NLRP3 , Caspase1 , ASC and IL⁃1β increased ( P <0. 05) . Compared with healthy tissue , the expressions of HAGLR and RUNX2 in TF mice decreased . Overexpression of HAGLR promoted the expressions of HAGLR and RUNX2 in TF mice , and increased the expression of MBV and tibia wet weight and IGF⁃1 (P < 0. 05) . Silencing RUNX2 on the basis of overexpression of HAGLR resulted in decreased expression of MBV , tibial wet weight and IGF⁃1 in TF mice (P < 0. 05) . However , the addition of NLRP3 inflammasome inhibitor MCC950 on top of the overexpression of HAGLR resulted in increased expressions of MBV , full⁃length tibia wet weight and IGF⁃1 (P < 0. 05) . Conclusion LncRNA HAGLR promotes the healing of tibial fractures by activating RUNX2 and inhibiting NLRP3 inflammasome .

To analyze the methodological and reporting quality of systematic reviews of prediction models published in Chinese journals, with the aim of providing reference for enhancing the overall quality of Chinese systematic reviews of prediction models.

BACKGROUND: Systemic lupus erythematosus (SLE) is a complex autoimmune disease, and the mechanism of SLE is yet to be fully elucidated. The aim of this study was to explore the role of two-pore segment channel 2 (TPCN2) in SLE pathogenesis. METHODS: Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the expression of TPCN2 in SLE. We performed a loss-of-function assay by lentiviral construct in Jurkat and THP-1 cell. Knockdown of TPCN2 were confirmed at the RNA level by qRT-PCR and protein level by Western blotting. Cell Count Kit-8 and flow cytometry were used to analyze the cell proliferation, apoptosis, and cell cycle of TPCN2-deficient cells. In addition, gene expression profile of TPCN2-deficient cells was analyzed by RNA sequencing (RNA-seq). RESULTS: TPCN2 knockdown with short hairpin RNA (shRNA)-mediated lentiviruses inhibited cell proliferation, and induced apoptosis and cell-cycle arrest of G2/M phase in both Jurkat and THP-1 cells. We analyzed the transcriptome of knockdown-TPCN2-Jurkat cells, and screened the differential genes, which were enriched for the G2/M checkpoint, complement, and interleukin-6-Janus kinase-signal transducer and activator of transcription pathways, as well as changes in levels of forkhead box O, phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of rapamycin, and T cell receptor pathways; moreover, TPCN2 significantly influenced cellular processes and biological regulation. CONCLUSION TPCN2 might be a potential protective factor against SLE.
Apoptosis/genetics* ; Cell Division ; Humans ; Jurkat Cells ; Lupus Erythematosus, Systemic/genetics* ; RNA, Small Interfering/genetics*

OBJECTIVE: To explore the genetic mechanism underlying a case with para-Bombay phenotype. METHODS: The ABO and Lewis phenotype were identified with serological methods. The coding regions of exons 6 and 7 of the ABO and FUT1 genes were amplified with PCR and directly sequenced. Haploid sequence analysis was carried out on the variant sites of the FUT1 gene. RESULTS: Serological analysis confirmed that the proband has a rare para-Bombay phenotype. Direct sequencing revealed that he was a B.01/O.01.02 heterozygote for the ABO gene, and had heterozygous deletion for the 768 and 881-882 sites of the FUT1 gene. Further haploid analysis showed that the c.881_882delTT deletion has occurred in one haploid while c.768delC was present in the other haploid. The proband was therefore determined as a FUT1*01N.13/01N.20 heterozygote, which have resulted in frameshift in polypeptide chain p.Phe294Cysfs*40 and p.Val257Phefs*23, respectively. CONCLUSION A rare bi-allelic heterozygous deletion of para-Bombay phenotype has been identified in a blood donor. The c.881_882delTT and c.768delC deletions may decrease the activity of α-1,2-fucosyltransferase.
Animals ; Male ; ABO Blood-Group System/genetics* ; Alleles ; Fucosyltransferases/genetics* ; Genotype ; Heterozygote ; Mutation ; Phenotype ; Humans

Objective To compare the application effect between the method of cassette automatic blood analyzer and the method of traditional serological .Methods Using the method of cassette automatic blood analyzer and the method of traditional serological , respectively ,the samples of blood donors in Dongguan from October 1 ,2013 to April 30 ,2014 were performed irregular antibody screening .The positive samples of screening were identified by using antiglobulin method ,and the irregular antibodies of blood do‐nors were analyzed .Results There were 95 positive cases of irregular antibody by the method of cassette automatic blood analyzer , and the detection rate was 0 .208% .There were 16 positive cases of irregular antibody by the method of traditional serological ,the detection rate was 0 .035% ,and there was statistical significance in differences (P< 0 .05) .The positive coincidence rate of the method of cassette automatic blood analyzer was 56 .547% ,higher than the rate of the method of traditional serological which was 28 .571 % (P<0 .05) .The positive rate of irregular antibody was 2 .242% in RhD‐negative blood donors ,higher than the rate in RhD‐positive blood donors which was 0 .198% (P<0 .05) .Conclusion The positive rate of irregular antibody in blood donors i‐dentified by using the method of cassette automatic blood analyzer is higher than the rate identified by using the method of tradition‐al serological .The irregular antibody screening should be performed for RhD‐negative blood donors .The types of irregular antibody in blood donors are mainly the types of IgM which are no clinical significance .