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Abstract Objective: Studies focusing on bone and joint infections (BJIs) in young infants are rare. Some cases of BJI are accompanied by sepsis. This study aimed to identify the clinical and bacteriological features of sepsis in neonates and young infants with BJIs. Methods: Neonates and infants younger than 3 months diagnosed with BJI in the present institution from 2014 to 2021 were retrospectively reviewed. Patient characteristics, clinical data, and outcomes were documented and compared between those with and without sepsis. Results: Twenty-five patients with a mean age of 34.8 days were included. Nine BJI cases had concomitant sepsis (group A), and 16 had BJI without sepsis (group B). Within group A, staphylococcus aureus was the major pathogenic germ (5 cases, of which 4 were of the methicillin-resistant staphylococcus aureus (MRSA) type). There was no statistical difference in male-to-female ratio, age, history of hospitalization, anemia, birth asphyxia, peripheral leukocyte counts, C-reactive protein on admission, and sequelae between groups. Univariate analyses indicated a significant difference in the incidence of septic arthritis (SA) combined with osteomyelitis (OM) (88.9% vs 37.5%), congenital deformities (44.4% vs 0%), and mean duration of symptoms (2.83 days vs 9.21 days) in comparisons between groups A and B. Conclusion: Staphylococcus aureus is the main pathogenic bacteria in BJI cases complicated with sepsis in neonates and young infants. Among infants younger than 3 months diagnosed with BJI, those with concurrent SA and OM, MRSA infection, or congenital deformities are more likely to develop sepsis.
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Tigliane type macrocyclic diterpenoids with special structures and diverse bioactivities are mainly extracted from plants of Euphorbiaceae and Thymelaeaceae. According to the different functional groups, they can be classified into types of phorbol esters, C-4 deoxyphorbol esters, C-12 deoxyphorbol esters, C-16 or C-17 substituted phorbol esters and others. Most of them present promising antiviral activities and cytotoxic activities and are expected to be developed as candidates for anti-AIDS, anti-tuberculosis, and anti-tumor clinical trials, demonstrating great potential for the application in healthcare. This paper reviews 115 novel tigliane-type diterpenoids discovered since 2013 and summarize their chemical structures and bioactivities, aiming to lay a foundation for further development and utilization of these compounds and provide new ideas for the development of clinical drugs.
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Phorbols , Molecular Structure , Diterpenes/chemistry , Antiviral Agents , Phorbol EstersABSTRACT
OBJECTIVE@#To assess the value of DNA methylation level of HYAL2 gene as a molecular marker for differential diagnosis of malignant and benign thyroid tumors.@*METHODS@#DNA methylation of HYAL2 gene in tissue specimens of 190 patients with papillary thyroid cancer (PTC) and 190 age- and gender-matched patients with benign thyroid tumors was examined by mass spectrometry, and the protein expression of HYAL2 was detected immunohistochemically for another 55 pairs of patients. Logistic regression analysis was performed to calculate the odds ratio (OR) and evaluate the correlation of per 10% reduction in DNA methylation with PTC. Receiver operating characteristic (ROC) curve analysis was performed and the area under curve (AUC) was calculated to assess the predictive value of alterations in HYAL2 methylation.@*RESULTS@#Hypomethylation of HYAL2_CpG_3 was significantly correlated with early-stage PTC (OR=1.51, P=0.001), even in stage I cancer (OR=1.42, P=0.007). Age-stratified analysis revealed a significantly stronger correlation between increased HYAL2_CpG_ 3 methylation and early-stage PTC in patients below 50 years than in those older than 50 years (OR: 1.89 vs 1.37, P < 0.05); ROC analysis also showed a larger AUC of 0.787 in younger patients. The results of immunohistochemistry showed that patients with PTC had significantly higher protein expressions of HYAL2 than patients with benign tumors.@*CONCLUSION@#The alterations of DNA methylation level of HYAL2 gene is significantly correlated with early-stage PTC, suggesting the value of DNA methylation level as a potential biomarker for differentiation of malignant from benign thyroid tumors.
Subject(s)
Humans , Middle Aged , Adenoma, Oxyphilic/genetics , Biomarkers, Tumor/metabolism , Cell Adhesion Molecules/metabolism , DNA Methylation , GPI-Linked Proteins/metabolism , Hyaluronoglucosaminidase/metabolism , Immunohistochemistry , Thyroid Cancer, Papillary/pathology , Thyroid Neoplasms/pathologyABSTRACT
Objective:To explore the effect of dyslipidemia on the clinical outcome of intracytoplasmic sperm injection-embryo transfer (ICSI-ET) in infertility patients receiving donor eggs.Methods:A total of 118 patients were selected to receive egg donors and ICSI-ET at the First Affiliated Hospital of Nanjing Medical University between April 2007 and December 2020. According to the levels of triacylglycerol, serum cholesterol, high density lipoprotein (HDL), and low density lipoprotein, they were divided into dyslipidemia group (35 cases) and normal blood lipids group (83 cases). The influence of body mass index (BMI) and age was adjusted by 1∶1 propensity score matching, and the general condition and clinical outcome of the two groups were analyzed retrospectively. Finally, the relationship between lipid composition and clinical outcome was analyzed according to patients′ age and BMI.Results:(1) Comparing the pre-matching dyslipidemia group with the normal blood lipids group, the BMI of the dyslipidemia group was significantly higher than that of the normal blood lipids group [(23.5±2.4) vs (22.4±2.7) kg/m 2], and the embryo implantation rate was significantly lower than that of the normal blood lipids group [13.6% (8/59) vs 27.3% (36/132)], the differences were statistically significant (both P<0.05). (2) There were no significant differences in years of infertility, number of pregnancies, number of abortions, number of transplanted embryos, protocol of endometrial preparation, endometrial thickness on transplantation day and high quality embryo rate between the two groups, through propensity score matching (all P>0.05). The biochemical pregnancy rate [28.6% (10/35)], embryo implantation rate [13.6% (8/59)] and live birth rate [20.0% (7/35)] in dyslipidemia group were significantly lower than those in the normal blood lipids group ( P<0.05). The clinical pregnancy rate was lower than that of the normal blood lipids group ( P>0.05). (3) The results of stratified analysis showed that the level of HDL in the clinically non-pregnant group was significantly lower than that in the pregnant group in patients ≤ 35 years old [(1.5±0.3) vs (1.8±0.5) mmol/L; P<0.05]. In the overweight recipient patients, the level of HDL of the clinically non-pregnant group was lower than that of the pregnant group ( P>0.05). Conclusions:Dyslipidemia significantly reduces the biochemical pregnancy rate, embryo implantation rate and live birth rate in patients with receiving donor eggs. Especially in patients aged ≤35 years old, the reduction of HDL is closely related to adverse pregnancy outcomes.
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ObjectiveTo observe the protective effect of Chaihu Jia Longgu Mulitang (CLMT) on dopaminergic neurons in Parkinson's disease with depression (PDD) model rats, and to explore the mechanism based on adenosine monophosphate-activated protein kinase/mammalian target of rapamycin (AMPK/mTOR) signaling pathway. MethodAmong the 80 male SD rats, 10 were randomly selected as normal group and the rest were treated with long-term low-dose subcutaneous injection of rotenone in the neck and back combined with chronic unpredictable mild stress (CUMS) to establish PDD rat model. The successfully modeled PDD rats were randomly divided into model group, western medicine group (madopar 0.032 g·kg-1+fluoxetine hydrochloride 0.002 g·kg-1), CLMT low-dose, medium-dose and high-dose groups (5, 10 and 20 g·kg-1), 10 rats in each group. Normal group and model group were administrated with the same amount of normal saline by gavage for 4 consecutive weeks. Behavioral changes of rats in each group were evaluated by open field test and pole climbing test. The content of dopamine (DA) and 5-hydroxytryptamine (5-HT) in cerebrospinal fluid was determined by high performance liquid chromatography (HPCL). The pathological changes of dopaminergic neurons in substantia nigra of rats were observed by hematoxylin-eosin (HE) staining. The positive expression of tyrosine hydroxylase (TH) and expression of α-synuclein in substantia nigra were detected by immunohistochemistry (IHC) and immunofluorescence (IF), repsectively. The protein expression of microtubule-associated protein 1 light chain 3 (LC3), adenosine monophosphate-activated protein kinase (AMPK), phosphorylated AMPK (p-AMPK), mammalian target of rapamycin (mTOR) and phosphorylated mTOR (p-mTOR) was detected by Western blot. ResultCompared with the conditions in normal group, the total horizontal distance and the activity time in the central region in open field test and the content of DA and 5-HT in cerebrospinal fluid were decreased (P<0.05, P<0.01), and the time of pole climbing was shortened (P<0.01), with increased score (P<0.01) in model group. Compared with model group, CLMT high-dose group and western medicine group increased the total horizontal distance and activity time in the central region and the content of DA and 5-HT (P<0.05, P<0.01), and extended the time of climbing pole (P<0.05), with decreased score (P<0.05, P<0.01). Compared with those in normal group, the number of dopaminergic neurons in the substantia nigra was reduced, with narrowed and loosely arranged cell body. The fluorescence expression of α-synuclein was enhanced (P<0.01), and the positive expression of TH was decreased (P<0.01) in model group. Compared with model group, CLMT high-dose group and western medicine group showed elevated number of dopaminergic neurons in the substantia nigra, with enlarged cell body, and decreased fluorescence expression of α-synuclein, and enhanced the positive expression of TH (P<0.05, P<0.01). Compared with normal group, model group had lowered expression of LC3Ⅱ/Ⅰ, p-AMPK/AMPK in striatum (P<0.05, P<0.01) and increased expression of p-mTOR/mTOR (P<0.01). Compared with those in model group, LC3Ⅱ/Ⅰ and p-AMPK/AMPK expression were increased (P<0.05, P<0.01) and p-mTOR /mTOR expression was decreased (P<0.01) in CLMT high-dose group and western medicine group. ConclusionCLMT exerts a neuroprotective effect by inhibiting rotenone neurotoxicity. It enhances the level of DA, and thus improves the depression condition in rats with Parkinson's disease. The underlying mechanism may be related to the regulation of AMPK/mTOR signaling pathway, activation of autophagy, and promotion of degrading α-synuclein.
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italic>Tripterygium wilfordii Hook. f. is a valuable medicinal plant, with anti-tumor, anti-inflammatory, immunosuppressive and other pharmacological activities. Triterpenoids are one of the main active components that exert pharmacological effects. However, the content of triterpenoids dominated by triptolide is very low in Tripterygium wilfordii, and the analysis of the biosynthetic pathway of triterpenoids in Tripterygium wilfordii provides an effective new idea for obtaining these compounds. 2,3-Oxidosqualene cyclases (OSCs) are the key enzyme that catalyzes the formation of triterpene skeleton diversity. Based on the genome and transcriptome data of Tripterygium wilfordii, 16 OSC genes were identified and analyzed. Phylogenetic analysis showed that 16 TwOSC proteins could be mainly classified as four groups. They are β-amyrin synthase group, friedelin synthase group, multifunctional amyrin synthase and cycloartenol synthase group. TwOSC6 was successfully cloned. Functional characterization analysis revealed that TwOSC6 can catalyze the formation of α-amyrin and β-amyrin. This indicates that TwOSC6 is a multifunctional amyrin synthase. This provides new gene resources for the diversity of Tripterygium wilfordii triterpenoids, as well as new gene elements for biosynthesis triterpenoids.
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Prostate cancer is one of the common malignant tumors of male urogenital system, and the incidence of prostate cancer in China has increased significantly in the past decade. At present, endocrine therapy based on androgen blockade is the main method of clinical treatment except radical surgery and radiotherapy/chemotherapy for prostate cancer. However, the clinical benefit can only be obtained in the early stage of treatment, and nearly 90% of patients will develop to the castration resistance, and among them, nearly 90% of patients will have bone metastasis. The quality of life decreases sharply with the progression of disease for patients. In addition to the androgen signal pathway, studies have shown that many other oncogenic signal pathways have involved in the development of castration resistance, including classic cancer signaling pathways, immune and inflammatory signaling pathways, etc. Understanding the mechanism of androgen independent signal pathway in the formation of castration resistance will help to understand the off-target effect of androgen blocking therapy and introduce new treatment targets or strategies to get rid of the "no drug available" dilemma for clinical treatment of castration resistance.
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Objective To investigate the karyotypes and C bands of Triatoma rubrofasciata in China, so as to understand its chromosome number, morphology and C-band staining of T. rubrofasciata. Methods The testis specimens were sampled from male T. rubrofasciata collected from Shunde City, Guangdong Province, prepared into slides of metaphase chromosomes and subjected to Giemsa staining and C-band staining. The morphology of metaphase chromosomes and the location of heterochromatin were observed using microscopy, and the long arm and short arm of each chromosome and total chromosome length were recorded to analyze the karyotypes and C bands of T. rubrofasciata. Results The male T. rubrofasciata presented a chromosome number of 2n = 25, including 22 autosomes and 3 sex chromosomes. The relative length of chromosomes ranged from 3.59% to 12.76%, the arm ratio was 1.06 to 1.24, and the centromere index was 44.76% to 48.47%. All chromosomes were metacentric chromosomes and the karyotype formula was 2n = 22 metacentric + X1X2Y, and the C bands varied on different chromosomes. No heterochromatin was found in the X chromosome, and the overall staining appeared pale, while heterochromatin was detected in all regions of the Y chromosome, and the overall staining appeared dark. In addition, heterochromatin was present in both ends of the autosome. Conclusions The male T. rubrofasciata presents a chromosome number of 2n = 25 in China, and the karyotype formula is 2n = 22 metacentric + X1X2Y. C-banding shows dark staining of the Y chromosome, pale staining of the X chromosome, and dark staining of both ends of the autosome. Our data may provide insights into the investigation on the origin, evolution and gene mapping of T. rubrofasciata in China.
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BACKGROUND@#Long non-coding RNA (lncRNA) actin filament-associated protein 1 antisense RNA 1 (AFAP1-AS1) functions as a competing endogenous RNA to regulate target genes expression by sponging microRNAs (miRs) to play cancer-promoting roles in cancer stem cells. However, the regulatory mechanism of AFAP1-AS1 in cervical cancer (CC) stem cells is unknown. The present study aimed to provide a new therapeutic target for the clinical treatment of CC.@*METHODS@#Hyaluronic acid receptor cluster of differentiation 44 variant exon 6 (CD44v6)(+) CC cells were isolated by flow cytometry (FCM). Small interfering RNAs of AFAP1-AS1 (siAFAP1-AS1) were transfected into the (CD44v6)(+) cells. The levels of AFAP1-AS1 were measured by quantitative real-time PCR (qRT-PCR). Sphere formation assay, cell cycle analysis, and Western blotting were used to detect the effect of siAFAP1-AS1. RNA pull-down and luciferase reporter assay were used to verify the relationship between miR-27b-3p and AFAP1-AS1 or vascular endothelial growth factor (VEGF)-C.@*RESULTS@#CD44v6(+) CC cells had remarkable stemness and a high level of AFAP1-AS1. However, AFAP1-AS1 knockdown with siAFAP1-AS1 suppressed the cell cycle transition of G(1)/S phase and inhibited self-renewal of CD44v6(+) CC cells, the levels of the stemness markers octamer-binding transcription factor 4 (OCT4), osteopontin (OPN), and cluster of differentiation 133 (CD133), and the epithelial-mesenchymal transition (EMT)-related proteins Twist1, matrix metalloprotease (MMP)-9, and VEGF-C. In the mechanism study, miR-27b-3p/VEGF-C signaling was demonstrated to be a key downstream of AFAP1-AS1 in the CD44v6(+) CC cells.@*CONCLUSIONS@#LncRNA AFAP1-AS1 knockdown inhibits the CC cell stemness by upregulating miR-27b-3p to suppress VEGF-C.
Subject(s)
Female , Humans , Cell Line, Tumor , Cell Proliferation/genetics , Gene Expression Regulation, Neoplastic , MicroRNAs/genetics , RNA, Long Noncoding/genetics , Uterine Cervical Neoplasms/genetics , Vascular Endothelial Growth Factor A/genetics , Vascular Endothelial Growth Factor CABSTRACT
Triptolide has wide clinical applications due to its anti-inflammatory, anti-tumor and immunosuppressive activities. In this study, we investigated the effect of blocking isopentenyl pyrophosphate (IPP) translocation on the biosynthesis of triptolide by exogenously adding D,L-glyceraldehyde (DLG) to the suspension cells of Ttripterygium wilfordii at different stages (7 d, 14 d). Subsequently, the cell viability, biomass accumulation, triptolide contents, as well as the profiles of the key enzyme genes involved in the upstream pathway of triptolide biosynthesis, were analyzed. The results showed that IPP translocation is involved in the biosynthesis of triptolide. IPP is mainly translocated from the plastid (containing the MEP pathway) to the cytoplasm (containing the MVA pathway) in the early stage of the culture, but reversed in the late stage. Blocking the translocation of IPP affected the expression of key enzyme genes involved in the upstream pathway of triptolide, which in turn affected the accumulation of triptolide. Understanding the characteristics and mechanism of IPP translocation provides a theoretical basis for further promoting triptolide biosynthesis through synthetic biology.
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Diterpenes , Epoxy Compounds , Hemiterpenes , Organophosphorus Compounds , PhenanthrenesABSTRACT
Objective This study aimed to determine the association of hyperlipidemia with clinical endpoints among hospitalized patients with COVID-19, especially those with pre-existing cardiovascular diseases (CVDs) and diabetes. Methods This multicenter retrospective cohort study included all patients who were hospitalized due to COVID-19 from 21 hospitals in Hubei province, China between December 31, 2019 and April 21, 2020. Patients who were aged < 18 or ≥ 85 years old, in pregnancy, with acute lethal organ injury (e.g., acute myocardial infarction, severe acute pancreatitis, acute stroke), hypothyroidism, malignant diseases, severe malnutrition, and those with normal lipid profile under lipid-lowering medicines (e.g., statin, niacin, fenofibrate, gemfibrozil, and ezetimibe) were excluded. Propensity score matching (PSM) analysis at 1:1 ratio was performed to minimize baseline differences between patient groups of hyperlipidemia and non-hyperlipidemia. PSM analyses with the same strategies were further conducted for the parameters of hyperlipidemia in patients with increased triglyceride (TG), increased low-density lipoprotein cholesterol (LDL-C), and decreased high-density lipoprotein cholesterol (HDL-C). Mixed-effect Cox model analysis was performed to investigate the associations of the 28-days all-cause deaths of COVID-19 patients with hyperlipidemia and the abnormalities of lipid parameters. The results were verified in male, female patients, and in patients with pre-existing CVDs and type 2 diabetes. Results Of 10 945 inpatients confirmed as COVID-19, there were 9822 inpatients included in the study, comprising 3513 (35.8%) cases without hyperlipidemia and 6309 (64.2%) cases with hyperlipidemia. Based on a mixed-effect Cox model after PSM at 1:1 ratio, hyperlipidemia was not associated with increased or decreased 28-day all-cause death [adjusted hazard ratio (
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , COVID-19/therapy , Cardiovascular Diseases/complications , Case-Control Studies , Cause of Death , China/epidemiology , Diabetes Mellitus, Type 2/complications , Hospitalization , Hyperlipidemias/complications , Propensity Score , Proportional Hazards Models , Retrospective Studies , Risk FactorsABSTRACT
Objective:To investigate the efficacy and pregnancy outcome of fertility-preserving treatment for patients with stage Ⅰa, grade 2 endometrial cancer (EC).Methods:Clinical data was retrospectively collected for EC or atypical endometrial hyperplasia (AEH) patients treated in Peking University People's Hospital, Foshan First People's Hospital of Guangdong Province and First Affiliated Hospital of Sun Yat-sen University, from 2010 to 2019. Inclusion criteria for fertility-preserving treatment included: (1) Age ≤45 years. (2) EC with histological differentiation of G 1, G 2 or endometrial AEH. (3) EC disease should be stage Ⅰa, confined to the endometrium without myometrial invasion, lymph node or extrauterine metastasis. Treatment regimen: patients were given oral progestin therapy and endometrial pathology was evaluated every three months. Patients were divided into three groups as G 2 EC group, G 1 EC group and AEH group based on the histological differentiation. Oncological and pregnancy outcomes were compared among them. Results:(1) Totally 57 eligible patients were included in this study, including 11 cases with G 2 EC, 22 cases with G 1 EC, and 24 cases with AEH. (2) Oncological outcome: among the three groups of G 2 EC, G 1 EC and AH, the complete remission rates (9/11, 91% and 96%, respectively) and recurrence rates (3/9, 30% and 22%, respectively) were not significantly different (all P>0.05). Median remission time was significantly longer in the G 2 EC group than those in the other two groups (8, 6 and 4 months; P=0.046). Among 9 G 2 EC patients who recurred after complete remission, three patients relapsed at 7, 18 and 53 months, respectively. All 3 patients chose fertility-sparing treatment again, and all achieved complete remission after retreatment. (3) Pregnancy outcome: among the three groups, the assisted reproduction technology rates (4/8, 5/18 and 36%, respectively) and pregnancy rates (6/8, 5/18 and 36%, respectively) had no significant difference ( P>0.05). However, time interval to pregnancy was shorter in G 2 EC patientsthan the other two groups (4, 9 and 22 months, respectively; P=0.006). Conclusions:Fertility-preserving treatment for patients with stageⅠa, G 2 endometrial cancer, may obtain a relatively high remission rate and an acceptable pregnancy rate. However, further exploration is needed due to the limited number of cases.
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Objective To analyze the changes in the endemic situation of schistosomiasis in national surveillance sites of Hunan Province, so as to provide scientific basis for the development of the schistosomiasis elimination programme in the province. Methods According to the requirements of the National Guidelines for Schistosomiasis Surveillance in China (2014 version), a total of 41 national schistosomiasis surveillance sites were assigned in all disease-endemic counties (districts) across Hunan Province. During the period between 2015 and 2019, Schistosoma japonicum infections were monitored in local residents, mobile populations and livestock, and snail status was monitored. The morbidity due to schistosomiasis and snail status was compared between years. Results The sero-prevalence of S. japonicum infections was 2.57% and 1.56% in local residents and mobile populations in national surveillance sites of Hunan Province from 2015 to 2019, respectively, and the sero-prevalence appeared a tendency towards a decline over years. A higher sero-prevalence rate of S. japonicum infections was seen in men than in women (P < 0.01). During the 5-year study period, the sero-prevalence rate of human S. japonicum infections appeared a tendency towards a decline in the marshland, embankment, inner embankment and hilly types of endemic areas over years. There were 44 and 19 egg -positives detected in local residents and 5 and 1 egg-positives in mobile populations in 2015 and 2016 respectively. A total of 9 346 domestic animals were monitored from 2015 to 2019, and 6 egg-positives were detected in 2015 and 2016 (all were bovine). A total of 0.155 billion m2 settings were surveyed from 2015 to 2019, and the mean density of living snails appeared a tendency towards a decline over years, with a 45.79% reduction in 2019 as compared to 2015. However, no S. japonicum infections were identified in snails during the 5-year period. A total of 1 469 mixed snail samples were detected using loop-mediated isothermal amplification (LAMP), and 6 positive snail samples were identified in 2015 (one sample) and 2017 (5 samples). Conclusions The overall endemic situation of schistosomiasis appears a tendency towards a decline in Hunan Province, and the prevalence of S. japonicum infections is at a low level in humans and livestock; however, there is still a risk of schistosomiasis transmission. Improvements of health education, intensification of schistosomiasis examinations in mobile populations and reinforcement of the surveillance-response system is required to consolidate the achievements of schistosomiasis control in Hunan Province.
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Objective To examine the construction and operation of the schistosomiasis diagnostic network platform in Hunan Province, so as to provide insights into the improvements of the schistosomiasis diagnostic network laboratory system in the province. Methods According to the criteria and requirements for the construction of the county-level schistosomiasis diagnostic network laboratory in China, the establishment and operation of the laboratory were assessed using self-assessment and field review in national schistosomiasis surveillance sites of Hunan Province. Results A total of 41 county-level schistosomiasis diagnostic network laboratories were built in national schistosomiasis surveillance sites of Hunan Province, and 36 met the requirements for the construction of the national schistosomiasis diagnostic network laboratory in China, which were approved for inclusion in the province-level schistosomiasis diagnostic network laboratory. During the six inter-laboratory comparisons performed by the national schistosomiasis diagnostic reference center of China, full consistency was achieved by 3 county-level schistosomiasis diagnostic network laboratories in Hunan Province, and the coincidence rates for re-review of serological and pathogenic detection samples were 98.40% and 100%, respectively. Conclusion The county-level schistosomiasis diagnostic network laboratory system has been preliminarily built and under effective operation in Hunan Province; however, the building capability remains to be improved.
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A method was established for simultaneous determination of 21 active constituents including flavanols, isoflavones, flavonols, dihydroflavones, dihydroflavonols, chalcones, pterocarpan, anthocyanidins and phenolic acids in Spatholobi Caulis by ultra fast liquid chromatography with triple quadrupole linear ion trap mass spectrometry(UFLC-QTRAP-MS/MS). Then, it was employed to analyze and evaluate the dynamic accumulation of multiple bioactive constituents in Spatholobi Caulis. The chromatographic separation was performed on a XBridge®C_(18)(4.6 mm×100 mm, 3.5 μm) at 30 ℃ with a gradient elution of 0.3% formic acid aqueous solution-methanol, and the flow rate was 0.8 mL·min~(-1), using multiple-reaction monitoring(MRM) mode. A comprehensive evaluation of the multiple bioactive constituents was carried out by gray correlation analysis(GRA). The 21 target components showed good linearity(r>0.999 0) in the range of the tested concentrations. The average recovery rates of the 21 components were from 97.46% to 103.6% with relative standard deviations less than 5.0%. There were differences in the contents of 21 components in Spatholobi Caulis at diffe-rent harvest periods. Spatholobi Caulis had high quality from early November to early December, which is consistent with the local tradi-tional harvest period. This study reveals the rule of the dynamic accumulation of 21 components in Spatholobi Caulis and provides basic information for the suitable harvest time. At the same time, it provides a new method reference for the comprehensive evaluation of the internal quality of Spatholobi Caulis.
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Chromatography, High Pressure Liquid , Fabaceae/chemistry , Phytochemicals/isolation & purification , Plant Stems/chemistry , Plants, Medicinal/chemistry , Tandem Mass SpectrometryABSTRACT
Molecular biology is a new subject that clarifies the phenomena and nature of life at the molecular level. Its development provides new biotechnology and methods for the study of traditional pharmacognosy. The formation of molecular biology has brought the development of pharmacognosy into a new era of gene research. Lonicerae Japonicae Flos is a classical Chinese medicine. Many scholars of home and abroad have carried out relevant studies on its molecular biology on the basis of the in-depth study with traditional methods, and have achieved certain results. In order to provide references on the method, technical for promoting the modernization of Lonicerae Japonicae Flos, and the development, protection, and utilization of other traditional Chinese medicine resources. This article summarized the application status of molecular biology methods and techniques on the identification, biosynthesis of active constituents, and molecular mechanism of secondary metabolite under stress conditions of Lonicerae Japonicae Flos in recent years. In hybridization technology of tag(RFLP), molecular markers based on PCR(RAPD, AFLP, SSR and ISSR), based on DNA sequence analysis of SNP and DNA barcode for the variety identification, diagnosis, identification of Lonicerae Japonicae Flos, and so forth in detail. At the same time, it is proposed that multi-omics technology can be used to build systems biology technology and platforms, and establish related models of secondary metabolite biosynthesis, so as to deepen acknowledge the molecular mechanism of the active component biosynthesis of Lonicerae Japonicae Flos and the accumulation of metabolites, life activities of other medicinal plants under adverse environment, then to regulate them.
Subject(s)
Amplified Fragment Length Polymorphism Analysis , Chromatography, High Pressure Liquid , DNA Barcoding, Taxonomic , Drugs, Chinese Herbal/pharmacology , Lonicera/chemistry , Medicine, Chinese Traditional , Microsatellite Repeats , Plants, Medicinal/chemistry , Polymorphism, Restriction Fragment Length , Polymorphism, Single Nucleotide , Random Amplified Polymorphic DNA Technique , Secondary MetabolismABSTRACT
Objective To compare the postoperative effects of Onlay transverse island preputial flap and Onlay transverse free island preputial graft for midshaft hypospadias repair.Methods Retrospective analysis of clinical data of 59 patients with midshaft hypospadias who underwent hypospadias repair from January 2013 to December 2018.Among them,49 children were repaired with Onlay transverse island preputial flap,aged from 14 months to 81 months,with the median age of 28 months.Ten children were repaired with Onlay transverse free island preputial graft.The age at surgery was from 15 months to 51 months,with the median age of 30 months.There was no significant difference in term of the age at surgery between the two groups (Z =-0.384,P =0.701).There were 9 cases of Donnahoo Ⅱ and 40 cases of Donnahoo Ⅲ in the traditional Onaly group,and 3 cases of Donnahoo Ⅱ and 7 cases of Donnahoo Ⅲ in the free graft Onlay group,respectively.There was no significant difference in the type of chordee between the two groups(x2 =0.161,P =0.688).The classification of hypospadias,the length of the urethroplasty and surgical complications were compared.The maximum urinary flow rate of some patients was summarized and analyzed after operation.Results Follow-up ranged from 3 to 72 months,with a median period of 38 months.In the children who underwent Onlay transverse island preputial flap,there were 2 cases of distal shaft,5 cases of middle shaft,and 42 cases of proximal shaft.The neourethra was 1.3-3.5 cm,with a median of 2.3 cm.There were 11 cases of fistula,1 case of glan dehiscence,and 1 case of diverticulum.Ten children were treated with Onlay transverse free island preputial graft.There were 1 case of distal shaft,2 cases of middle shaft and 7 cases of proximal shaft.The neourethra was 1.2-2.5 cm,with a median of 2.0 cm.Two cases of fistula occurred after operation.There was no significant difference in hypospadias classification between the two groups(x2 =1.313,P =0.519).There was no significant difference in the length of the urethra between the two groups (Z =-1.732,P =0.083).There was no significant difference in the incidence of postoperative complications between the two surgical methods (x2 =0.001,P =0.973).The patients' parents were satisfied with the appearance of genital organ after operation.Fourteen patients finished the uroflowmetry 3 months after surgery.Twelve patients in the traditional Onlay group had a urine volume of 60.2-147.8 ml,with an average of 98.8 ml.The maximum urinary flow rate was 3.5-13.6 ml/s,with an average of 8.1 ml/s.In the free graft Onlay group,the urine volume was 101.3-165.4 ml,with an average of 133.5 ml,and the maximum urinary flow rate was 9.6-15.7 ml/s,with an average of 12.7 ml/s.Conclusions Onlay transverse free island preputial graft has the same complication incidence with Onlay transverse island preputial flap,but could get a good maximum urinary flow rate and cosmetic result,which could achieve a good choice for midshaft hypospadias repair.
ABSTRACT
A method, for determination of saponins, amino acids and nucleosides in Panacis Japonici Rhizoma of ultra fast liquid chromatography with triple quadrupole linear ion trap mass spectrometry (UFLC-QTRAP-MS/MS), was established to investigate the effect of different processing methods on the target components of Panacis Japonici Rhizoma. The chromatographic separation was performed on a XBridgeC₁₈(4.6 mm×100 mm, 3.5 μm) at 30 °C with a gradient elution of 0.1% formic acid solution-0.1% formic acid acetonitrile, and the flow rate was 0.8 mL·min⁻¹, using multiple-reaction monitoring (MRM) mode. The grey relational analysis was adopted for the analysis of different processing samples. The results showed that the thirty-three constituents were in a good linear range and the correlation coefficient was greater than 0.999 0; the precision, repeatability and stability were good; the average recovery rates were between 95.33% and 101.8%, and the relative standard deviations were less than 5%. The result of grey relational analysis showed that the complete rhizomes without peeling, which were adopted for the microwave dried method, had the best quality. The established method was accurate and reliable, which could be used to appraise the quality of Panacis Japonici Rhizoma. Our study may lay the way for the processing method of Panacis Japonici Rhizoma in optimization,normalization and standardization.
Subject(s)
Amino Acids , Chromatography, High Pressure Liquid , Chromatography, Liquid , Nucleosides , Panax , Chemistry , Phytochemicals , Rhizome , Chemistry , Saponins , Tandem Mass SpectrometryABSTRACT
An analytical method based on UFLC-QTRAP-MS/MS was established for simultaneous determination of thirty-three components including steroidal saponins, homoisoflavonoids, amino acids and nucleosides in Ophiopogonis Radix. Thirty-three target components of commercial medicinal materials of Maidong were comparative analysis. Synergi™ Hydro-RP 100 column (2.0 mm × 100 mm, 2.5 μm) was used with 0.1% formic acid solution-0.1% formic acid acetonitrile for gradient elution at a flow rate of 0.4 mL·min⁻¹. In addition, multiple reaction monitoring (MRM) mode was employed. The data were comprehensively processed and analyzed with hierarchical clustering analysis(HCA), principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA) methods. All components showed good linearity(>0.999 0) within the tested ranges. The average recoveries were between 96.23%-102.0%, and the relative standard deviation(RSD) were less than 5%. The results showed that there were significant differences in components between Ophiopogonis Radix and Liriopes Radix, with seven components obviously different. This method was useful for providing basis for the comprehensive evaluation and intrinsic quality control of Ophiopogonis Radix and Liriopes Radix , and may provide a new method reference for the identification of Ophiopogonis Radix and Liriopes Radix.
Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Liriope Plant , Chemistry , Ophiopogon , Chemistry , Phytochemicals , Plant Roots , Chemistry , Saponins , Tandem Mass SpectrometryABSTRACT
Dao-di herbs have been recognized as "quality models" with a firmly stable status. The formation of Dao-di herbs quality is involved from the genetic inheritance on the molecular level to the metabolic phenotype of final products, and the full material-based biosynthetic pathway remains unknown. In recent years, an increasing variety of omics technologies has provided new methods and ideas for the analysis of complex life systems and are suitable for explanation of quality formation in Dao-di herbs as well. In order to alleviate the scarcity of natural resources and offer scientific guidance of transplanting varieties, achievements of omics in the aspects of Dao-di herbs from genetics to phenotyping, the biosynthetic pathway of secondary metabolites, the interaction with human body and the new methods of quality evaluation have been summarized. It will be a fundamental work for protection and utilization of Chinese medicine resources.