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Objective:To analyze the feasibility of fluorescence staining in the detection of Demodex mites. Methods:A single-center split-face study was conducted, and patients with clinically diagnosed rosacea or seborrheic dermatitis were enrolled from the Department of Dermatology, Southern University of Science and Technology Hospital from October 2020 to June 2021. Samples were obtained from the patients′ cheeks by using the squeeze-adhesion method, and Demodex mites were detected by fluorescence staining and direct microscopic examination separately. The detection rate, number of detected Demodex mites, and time for reading slides were compared between the above two detection methods, and the detection rate and number of detected Demodex mites were further compared between the fluorescence staining and standardized skin surface biopsy (SSSB). Chi-square test was used to compare enumeration data, and paired t-test for measurement data. Results:A total of 433 volunteers aged 28.3 ± 3.5 years were enrolled, including 185 males and 248 females. The performance of fluorescence staining was compared with that of direct microscopic examination in 338 pairs of samples obtained by the squeeze-adhesion method, and compared with that of SSSB in 95 pairs of samples obtained by the squeeze-adhesion method. The detection rate of Demodex mites by fluorescence staining was significantly higher (34.0%, 115/338) than that by direct microscopic examination (31.4%, 106/338; McNemar test, P = 0.004) ; among 118 positive samples, the number of detected Demodex mites by fluorescence staining ([8.0 ± 3.3]/cm 2) was also significantly higher than that by direct microscopic examination ([5.5 ± 2.9]/cm 2, t = 9.21, P < 0.001) ; the time for reading slides undergoing fluorescence staining was significantly shorter (8.3 ± 1.2 minutes) than that undergoing direct microscopic examination (17.3 ± 2.5 minutes, t = 38.44, P < 0.001) ; there was favorable consistency in fluorescence staining results between two clinical laboratorians (kappa value = 0.935, P < 0.001). The detection rate of Demodex mites by fluorescence staining (34.7%, 33/95) was higher than that by SSSB (33.7%, 32/95; McNemar test, P < 0.001) ; among 35 positive samples, the number of detected Demodex mites by fluorescence staining was also significantly higher ([11.4 ± 4.2]/cm 2) than that by SSSB ([9.8 ± 4.8]/cm 2; t = 4.77, P < 0.001) . Conclusion:Compared with direct microscopic examination and SSSB, fluorescence staining was more sensitive in the detection of Demodex mites, with better consistency between different observers and shorter time for reading slides.
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The many-banded krait, Bungarus multicinctus, has been recorded as the animal resource of JinQianBaiHuaShe in the Chinese Pharmacopoeia. Characterization of its venoms classified chief phyla of modern animal neurotoxins. However, the evolutionary origin and diversification of its neurotoxins as well as biosynthesis of its active compounds remain largely unknown due to the lack of its high-quality genome. Here, we present the 1.58 Gbp genome of B. multicinctus assembled into 18 chromosomes with contig/scaffold N50 of 7.53 Mbp/149.8 Mbp. Major bungarotoxin-coding genes were clustered within genome by family and found to be associated with ancient local duplications. The truncation of glycosylphosphatidylinositol anchor in the 3'-terminal of a LY6E paralog released modern three-finger toxins (3FTxs) from membrane tethering before the Colubroidea divergence. Subsequent expansion and mutations diversified and recruited these 3FTxs. After the cobra/krait divergence, the modern unit-B of β-bungarotoxin emerged with an extra cysteine residue. A subsequent point substitution in unit-A enabled the β-bungarotoxin covalent linkage. The B. multicinctus gene expression, chromatin topological organization, and histone modification characteristics were featured by transcriptome, proteome, chromatin conformation capture sequencing, and ChIP-seq. The results highlighted that venom production was under a sophisticated regulation. Our findings provide new insights into snake neurotoxin research, meanwhile will facilitate antivenom development, toxin-driven drug discovery and the quality control of JinQianBaiHuaShe.
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Homocestkink is a mktabolitk of mkthionink,which has bkkn provkd to bk associatkd with multiplk sestkmic diskasks. In pkdiatric diskasks,homocestkinkmia has bkkn paid mork and mork attkntion in rkcknt ekars. Hepkr-homocestkinkmia is associatkd with somk diskasks of cardiovascular sestkm,rhkumatoid immunk sestkm,urinare sestkm, nkrvous sestkm,blood sestkm,digkstivk sestkm in childrkn. Lnd karle mkasurkmknt of homocestkinkmia plaes an impor-tant rolk in thk diagnosis,trkatmknt and prkvkntion of somk diskasks in childrkn. Through sunmmarizing thk mktacha-nism of homocestkink and its rklationship with multiplk sestkmic diskasks in childrkn,wk invkstigatk thk clinical valuk of homocestkink in childrkn.
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A rapid and simple method for the determination of cyanide in blood was developed based on pinhole shell-isolated nanoparticles (pinSHINs)-enhanced Raman spectroscopy and an online lysis-purging and trapping approach.In the online lysis-purging and trapping device, the bound cyanide in blood can be cleaved through sulfuric acid acidification, and transferred into HCN volatile gas, then purged into alkaline solution to form NaCN solution, thus high-efficient liberation and entrapment of cyanide from the methemoglobin-bound form can be achieved.The pinSHINs substrate is quite stable to weaken the gold-dissolution effect caused by cyanide under strong alkaline condition, and therefore the detection window can be prolonged to 1 h comparing with 5 min of AuNPs.A limit of detection down to 10 μg/L and a linear range from 100-2000 μg/L in blood were achieved in this method.This method was further applied to rapid measurement of blood samples of cyanide exposed rats and clinic poisoned patients, which provided a sensitive, selective and reliable way for rapid detection of cyanide poisoning.
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The multidisciplinary synthetic therapy for the colorectal liver metastases has been a hot spot in clinical research,which includes operative therapy,tumor local therapy,conversion therapy,chemotherapy, molecular targeted therapy and so on. It is need to choose multiple therapies for the patients and make the whole treatment strategy in accordance with the condition of patients to maximize the survival benefit in clinical prac-tice. So,it is important to comprehend the newest research process of the clinical therapy to make a good choice for the colorectal liver metastases patients.
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Objective To explore the inhibitory effect of tetramethylpyrazine (TMP) on ultraviolet A-induced senescence as well as matrix metalloproteinase-1 (MMP-1) and-3 (MMP-3) mRNA expressions in human dermal fibroblasts (HDFs).Methods HDFs were isolated from the prepuce by enzymatic digestion, and subjected to primary culture.Cultured HDFs were randomly divided into several groups: control group cultured in high-glucose DMEM medium and receiving no treatment, three TMP groups treated with 20, 50 and 100 mg/L TMP respectively, UVA group receiving UVA radiation alone, UVA + TMP groups pretreated with 20, 50 and 100 mg/L TMP respectively for different durations followed by UVA radiation.UVA radiation was given once daily for 5 consecutive days.The 55th passage HDFs served as the P55 group (senescence control group).Subsequently, CCK-8 assay was performed to evaluate the proliferative activity of HDFs in vitro, optical microscopy to observe the morphologic changes of HDFs after UVA radiation, β-galactosidase staining to estimate the senescence in HDFs, and real-time fluorescence-based quantitative PCR to quantify the mRNA expressions of MMP-1 and MMP-3 in HDFs.Statistical analysis was carried out by one-way analysis of variance (ANOVA) followed by least significant difference (LSD)-t test or Dunnett's T3 test.Results Compared with the control group, the proliferation of HDFs was significantly but transiently inhibited in vitro after the treatment with 100 mg/L TMP for 48 hours (P ＜ 0.05), but showed no significant changes after the treatment with 20 or 50 mg/L TMP for 24, 48 or 72 hours or after the treatment with 100 mg/L TMP for 24 or 72 hours (all P ＜ 0.05).The pretreatments with TMP of 20, 50 and 100 mg/L for 24, 48 and 72 hours all promoted the proliferation of HDFs to a certain degree in the UVA + TMP groups compared with the UVA group, with significant differences in cellular proliferative activity among the UVA group, UVA + TMP groups and control group at 24, 48 and 72 hours (F =17.451,15.231, 23.535, all P ＜ 0.01).Compared with the UVA group, the proliferative activity of HDFs was significantly increased in UVA + 100-mg/L TMP group at 24, 48, 72 hours, UVA + 50-mg/L TMP group at 24 and 72 hours and UVA + 20-mg/L TMP group at 72 hours.After repetitive UVA radiation, HDFs in the UVA group experienced an increase in cell volume, granule acount, and β-galactosidase expression, which was similar to the changes in the P55 group, while the pretreatments with 20, 50 and 100 mg/L TMP for 24 hours suppressed these UVA-induced changes in HDFs.The percentage of β-galactosidase-positive HDFs was 68.417％ ± 1.181％ in the UVA group, 58.167％ ± 5.620％ in the UVA + 20-mg/L TMP group, 45.167％ ± 5.502％ in the UVA + 50-mg/L TMP group, 43.000％ ± 2.000％ in the UVA + 100-mg/L TMP group, 33.667％ ± 5.865％ in the control group, and 76.000％ ± 6.557％ in the P55 group, with significant differences among these groups (F =45.918, P ＜ 0.01).Furthermore, the UVA group significantly differed from the UVA + TMP groups and control group in the percentage of β-galactosidase-positive HDFs and mRNA expressions of MMP-1 and MMP-3 (all P ＜ 0.05).Conclusion TMP can protect HDFs against senescence induced by repetitive UVA radiation, and down-regulate the mRNA expressions of MMP-1 and MMP-3 during senescence.
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JAK/STAT includes many types of cell factors. Growth factor signal transduction is an important pathway and widely participates in cell multiplication, differentiation, and immunity adjustment. Aberrant-activated JAK/STAT signaling pathway is in-volved in carcinogenesis. Recent studies demonstrated that abnormal expression and activation of STAT3 were found in lymphoma. Constitutive activation of STAT3 promotes development, invasion, and metastasis of cancer. AG490, a JAK2 inhibitor, can block the JAK/STAT3 signal pathway and reduce the pathway's downstream STAT3 expression. Several studies showed that AG490 can inhibit cell proliferation and promote apoptosis in lymphoma. AG490 combined with chemotherapy drugs could improve sensitivity. In our study, we reviewed the potential role of JAK/STAT signaling pathway and the blocker AG490 in lymphoma.
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Codon usage bias is an important characteristic of genetic information transfer in organisms. Analysis of codon usage bias of different species is important for understanding the rules on genetic information transfer. The previous method for analysis of codon usage bias is mainly based on genomic data. However, this method is greatly limited, because the genome sequences of higher organisms are still not available up to now. In this study, we found that we could obtain the same optimal codons of Ganoderma lucidum (Curtis: Fr.) P. Karst based on its whole genomic data or large-scale transcriptomic data from its liquid-cultured hyphae, primordium and fruiting body, separately. This result indicated the feasibility to understand the codon usage bias based on the large-scale transcriptomic data. By calculating the proportion of rare codons of Escherichia coli and Saccharomyces cerevisiae in 26 terpene synthases (TS) of G. lucidum, we found that the rare codons of S. cerevisiae have a higher proportion in TS genes, while the rare codons of E. coli have relatively lower, suggesting that the TS genes of G. lucidum are possibly more difficult to be expressed in S. cerevisiae than in E. coli. Chemical synthesis of TS genes according to the yeast optimal codons will be an effective way to solve the problem on the mismatch of gene codon bias between the foreign genes and the host strain.
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Objective To evaluate the value of PET/CT in preoperative assessment and postoperative monitoring of ovarian cancer. Methods A retrospective analysis was conducted in 45 patients of ovarian neoplasm with clinical records underwent 18F-FDG PET/CT, including 10 patients underwent PET/CT before surgery and 35 patients after surgery. The clinical follow-up time was 6 months at least. The diagnosis based on pathology and clinical follow-up data. Results (1) The sensitivity, specificity and accuracy of PET/CT in detecting ovarian cancer were 94.6%,75.0%and 91.1%, respectively. (2) Ten patients before surgery were all detected tumor by PET/CT, but 2 of them were false positive based on pathologic results. (3) Two patients with non-standard surgery were detected tumor by PET/CT. In 33 patients after standard surgery, 6 patients were no tumor detected by PET/CT. In addition,4 patients with normal CA125 and no signs of recurrence and metastasis were detected tumor by PET/CT. The pathology and clinical follow-up data supported the results. 23 patients with higher CA125 were diagnosed recurrence and metastasis based on pathology and clinical follow-up data, 21 of them were detected tumor by PET/CT. Conclusion 18F-FDG PET/CT plays an important role in preoperative assessment, early diagnosis and accurate positioning of recurrent and metastasis of ovarian cancer. It can be used to guide the clinical treatment.
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Codon is an important carrier of genetic information in organisms. In evolution, codon bias is formed in many organisms. In this study, the analysis of expressed genes in Lonicera japonica Thunb. is presented. The codon bias of genes with complete coding region is analyzed through CodonW. And we calculated the codon usage bias of hydroxycinnamoyl-CoA: quinate hydroxycinnamoyl transferase (HQT) genes. The results showed that L. japonica were bias toward the synonymous codons with A and T at the third codon position.
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ERF family transcription factor (TF) represented ethylene-responsive protein which harbored a conserved AP2 domain. After searching the plant transcription factor database, a total of 75 unigenes was found which contained AP2 domain from the transcriptome dataset of Panax quinquefolius L. One unique sequence of ERF transcript, named as PqERF1, was cloned with entire open reading frame of 933 base pairs (bp). Protein prediction result indicated that the gene was localized in nucleus and had a conserved AP2 domain. PqERF1 gene could be induced by methyl jasmonate (MeJA) which was consistent to the inducing profile of triterpene ginsenosides. InterproScan prediction indicated that PqERF1 was probably a pathogenesis-related gene. Sequence alignment and phylogenetic analysis demonstrated PqERF1 was with high identity and had relative close relationship to the NtERF4 (Nicotiana tabacum), PhERF12 (Petunia x hybrida) and DcERF1 (Daucus carota) which was related to plant defense, regulation of secondary metabolism and the flower senescence respectively. Therefore, the gene was likely involved in regulation of secondary metabolism, plant defense and physical processes which would provide gene resource for further study on secondary metabolite synthesis and molecular breeding of P. quinquefolius.
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DNA barcoding is a technique in which species identification and discovery are performed by using short and standard fragments of DNA sequences. In this study, eleven species of Paris, including seven varieties, were sampled. Five chloroplast sequences, psbA-trnH, rpoB, rpoC1, rbcL, matK, and one nuclear marker, the second internal transcribed spacer (ITS2) of ribosomal DNA, were amplified and sequenced. The PCR amplification and sequencing efficiency, intra- and inter-specific divergence and barcoding gap were used to evaluate different loci, and the identification efficiency was assessed using BLAST1 and Nearest Distance methods. The ITS2 sequences in the studied samples of Paris were amplified and sequenced successfully using primers designed by our group, while matK showed low level in the amplification and psbA-trnH was difficult for sequencing because of over 800 bp and poly (A) structure. Analysis of the intra- and inter-specific divergence and barcoding gap showed ITS2 was superior to other loci. The ITS2 showed a much higher percentage of success (100%) in identification than other five loci, none of which indicated more than 50% except matK (52.9%). The 2-locus combination of rbcL+matK didn't improve ability of authentication. In addition, the rate of successful identification with ITS2 kept 100% when the samples were expanded to 67 samples of 29 species. In conclusion, ITS2 can be used to correctly identify medicinal plants of Paris, and it will be a potential DNA barcode for identifying medicinal plants of other taxa.
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The dried succulent stems of Cistanche (Cistanche deserticola Y. C. Ma and Cistanche tubulosa Wight.) are one of the most widely used components of traditional Chinese medicines. However, it is often confused and substituted with the roots of Orobanche pycnostachya, Boschniakia rossica (Cham. & Schltdl.) Standl., Cistanche sinensis Beck, and Cistanche salsa (C. A. Mey.) Beck. In this study, we identified psbA-trnH regions from species and tested their suitable for the identification of the above mentioned taxa. The psbA-trnH sequences showed considerable variations between species and thus were revealed as a promising candidate for barcoding of Cistanche species. Additionally, the average genetic distance of psbA-trnH ranging from 0.077% to 0.743%. In contrast, the intra-specific variation among Cistanche species was found to be significantly different from those of other species, with percentages of variation studied ranged from 0% to 0.007%. The sequence difference between the psbA-trnH sequences of Cistanche species and Orobanche pycnostachya ranged from 0.979% to 1.149%. The distance between the Cistanche species and Boschniakia rossica ranged from 1.066% to 1.224%. Our results suggest that the psbA-trnH intergenic spacer region represent a barcode that can be used to identify Cistanche species and other morphologically undistinguishable species.
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Background and purpose:A pressing obstacle in clinical chemotherapy is drug resistance in breast cancer.A nano-delivery system,which has many advantages as a drug carrier,such as carrying anticancer drugs,can be used effectively to overcome drug resistance in tumors.This paper examined a new nano-delivery system,called calcium phosphate and glycerophosphocholine-mPEG(CAP/GPC-MPEG)composite nanoparticle and its influence on the cellular drug uptake of BCRP-over expressing mitoxantrone(MIT)-resistant breast cancer cell MCF-7/MIT.This paper will also examine its effect on overcoming drug resistance in the MCF-7/MIT cells.Methods:After the calcium phosphate and GPC-mPEG composite nanoparticles were designed and prepared,the entrapment efficiency and in vitro drug release of mitoxantrone-loaded nanoparticles were investigated.Quantitative comparisons were made between cellular uptake of drug-loaded nanoparticles and free drugs.Finally,a confocal laser scanning microscopy Was used to compare the subcellular distribution of drug-loaded nanoparticles and the free drugs.Results:Calcium phosphate and GPC-mPEG composite nanoparticles were nanoporous spherical particles with diameters between 50-100 mn.The MIT-loaded nanoparticles have an entrapment efficiency of(89.45±0.05)%.Although the drug-loaded nanoparticles showed an initial burst of drug release,it was followed by a more sustained release.The concentration of mitoxantrone was 1.89 times treated with MIT-loaded nanoparticles for 1 h compared to that treated with free mitoxantrone for 1 h in MCF-7/MIT cells.and which was 2.33 times in MCF-7 cells.Fluorescent red mitoxantrone appeared in the cytoplasm and nucleus of the MCF-7 and MCF-7,MIT cells treated with MlT-loaded nanoparticles whereas it is almost undetected in both cells treated with free mitoxantrone.Conclusion:Calcium phosphate and GPC-mPEG composite nanoparticles Can promote the cellular uptake and entering of mitoxantrone to the nucleus in MCF-7 and its corresponding BCRP-over expressing MIT-resistant MCF-7/MIT breast cancer cell lines.This nanoparticle is a potential nano-carrier for overcoming drug resistance in tumors.
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Gastric cancer is one of the most common malignant tumors. Traditional Chinese medicine (TCM) has been widely used in treatment of gastric cancer, but still lacking large sample controlled trial to evaluate its efficacy.
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The incidence of colorectal cancer is high among the elderly. Traditional Chinese medicine (TCM) has been widely used in the treatment for colorectal cancer of old people. However, controlled trials with large sample size evaluating the effect of TCM are rare.