ABSTRACT
Objective To evaluate the performance of recombinase-aided amplification (RAA) assay in detection of Clonorchis sinensis metacercariae in freshwater fish samples, so as to provide insights into standardization and field application of this assay. Methods Wild freshwater fish samples were collected in the rivers of administrative villages where C. sinensis-infected residents lived in Jiangyan District, Xinghua County and Taixing County of Taizhou City, Jiangsu Province from June to September 2022. Genomic DNA was extracted from six freshwater fish specimens (5 g each) containing 0, 1, 2, 4, 8 and 16 C. sinensis metacercariae for fluorescent RAA assay, and the diagnostic sensitivity was evaluated. Fluorescent RAA assay was performed with genomic DNA from C. sinensis, Metorchis orientalis, Haplorchis pumilio and Centrocestus formosanus metacercariae as templates to evaluate its cross-reactions. In addition, the detection of fluorescent RAA assay and direct compression method for C. sinensis metacercariae was compared in field-collected freshwater fish samples. Results Positive amplification was found in fresh-water fish specimens containing different numbers of C. sinensis metacercariae, and fluorescent RAA assay was effective to detect one C. sinensis metacercaria in 5 g freshwater fish specimens within 20 min. Fluorescent RAA assay tested negative for DNA from M. orientalis, H. pumilio and C. formosanus metacercariae. Fluorescent RAA assay and direct compression method showed 5.36% (93/1 735) and 2.88% (50/1 735) detection rates for C. sinensis metacercariae in 1 735 field-collected freshwater fish samples, with a statistically significant difference seen (χ2 = 478.150, P < 0.001). There was a significant difference in the detection of C. sinensis metacercariae in different species of freshwater fish by both the direct compression method (χ2 = 11.20, P < 0.05) and fluorescent RAA assay (χ2 = 20.26, P < 0.001), and the detection of C. sinensis metacercariae was higher in Pseudorasbora parva than in other fish species by both the direct compression method and fluorescent RAA assay (both P values < 0.05). Conclusions Fluorescent RAA assay has a high sensitivity for detection of C. sinensis metacercariae in freshwater fish samples, and has no cross-reactions with M. orientalis, H. pumilio or C. formosanus metacercariae. Fluorescent RAA assay shows a higher accuracy for detection of C. sinensis infections in field-collected freshwater fish than the direct compression method.
ABSTRACT
Tooth germ injury can lead to abnormal tooth development and even tooth loss, affecting various aspects of the stomatognathic system including form, function, and appearance. However, the research about tooth germ injury model on cellular and molecule mechanism of tooth germ repair is still very limited. Therefore, it is of great importance for the prevention and treatment of tooth germ injury to study the important mechanism of tooth germ repair by a tooth germ injury model. Here, we constructed a Tg(dlx2b:Dendra2-NTR) transgenic line that labeled tooth germ specifically. Taking advantage of the NTR/Mtz system, the dlx2b+ tooth germ cells were depleted by Mtz effectively. The process of tooth germ repair was evaluated by antibody staining, in situ hybridization, EdU staining and alizarin red staining. The severely injured tooth germ was repaired in several days after Mtz treatment was stopped. In the early stage of tooth germ repair, the expression of phosphorylated 4E-BP1 was increased, indicating that mTORC1 is activated. Inhibition of mTORC1 signaling in vitro or knockdown of mTORC1 signaling in vivo could inhibit the repair of injured tooth germ. Normally, mouse incisors were repaired after damage, but inhibition/promotion of mTORC1 signaling inhibited/promoted this repair progress. Overall, we are the first to construct a stable and repeatable repair model of severe tooth germ injury, and our results reveal that mTORC1 signaling plays a crucial role during tooth germ repair, providing a potential target for clinical treatment of tooth germ injury.
Subject(s)
Animals , Mice , Mechanistic Target of Rapamycin Complex 1/pharmacology , Signal Transduction , Tooth/metabolism , Tooth Germ/metabolism , OdontogenesisABSTRACT
Objective:To investigate the clinical value of three-dimensional arterial spin marker imaging(3D-ASL) for evaluating collateral circulation compensatory status in patients with cerebral infarction.Methods:A total of 24 patients with cerebral infarction who were treated at Wenzhou Hospital of Traditional Chinese Medicine Affiliated to Zhejiang University of Chinese Medicine from September 2017 to January 2019 were examined by 1.5-T high-field MR scanner.The changes of cerebral blood flow(CBF) between the infarcted area and the contralateral normal area were compared when the labeled delay time(PLD)=1.5s and PLD=2.5s, and the patients were divided into the group with strong collateral circulation compensation and the group with weak collateral circulation compensation according to the difference in lesion area measured by 3D-ASL(PLD=1.5s and 2.5s). The changes of national institutes of health stroke scale(NIHSS) score and Bathel index at admission and 15 days after admission were compared.Results:3D-ASL measurement of PLD=1.5s showed that the CBF value in the infarcted area was (33.70±20.83)mL/(100g×min), which was significantly lower than that in the contralateral normal area(PLD=1.5s)[(49.93±13.13)mL/(100g×min)]( t=3.229, P<0.05). When PLD=2.5s, ASL measurement results showed that the CBF value in the infarcted area increased significantly[(58.26±23.50)mL/(100g×min) vs.(33.70±20.83)mL/(100g×min)]( t=3.831, P<0.05), and the CBF value in the contralateral normal area also increased significantly[(68.29±14.03)mL/(100g×min) vs.(49.93±13.13)mL/(100g×min)]( t=4.681, P<0.05). At this time, the CBF value in the infarcted area was still significantly lower than that in the contralateral normal area[(58.26±23.50)mL/(100g×min) vs.(68.29±14.03)mL/(100g×min)]( t=1.795, P<0.05). On the day of admission, the BI index of patients in the weak collateral circulation compensation group(12 cases) was significantly higher than that in the strong collateral circulation compensation group(12 cases)[(90.42±10.50) vs.(67.92±27.57)]( t=2.642, P<0.05), and the NIHSS score was significantly lower than that in the strong collateral circulation compensation group[(1.25±1.01) vs.(3.83±3.62)]( t=2.378, P<0.05). After 15 d of admission, there were no significant differences in BI index and NIHSS score between the two groups(all P>0.05). Conclusion:3D-ASL with different delay time can effectively and intuitively reflect collateral circulation compensation of patients with cerebral infarction, and it has very important reference value for the evaluation of patients' disease and the formulation of clinical treatment plan.
ABSTRACT
The aim of this study was to evaluate the effects of compound probiotics on milk performance,blood biochemical indexes,rumen fermentation parameters and nutrient digestibility of Holstein dairy cows in middle-and-later lactation.Forty-eight dairy cows with similar milk yield,days in milk and age were randomly divided into 4 groups treated with four different levels of compound probiotics as follows:the control group (0 g/d),group 1 (10 g/d),group 2 (20 g/d) and group 3 (30 g/d).The adaptation period was 7 days,and the total experimental period was 60 days.Results showed that:compared with the control group,supplementation of compound probiotics could improve milk yield and significantly increase the concentration of milk protein,lactose and total solids (P＜0.05);there was no significant effect on blood biochemical indexes (P＞ 0.05);compound probiotics could significantly increase the content of ammonia and microbial protein content (MCP) in the rumen (P＜ 0.05);the apparent nutrient digestibility of crude protein and crude fat of group 2 was significantly higher than other groups (P＜0.05);compared with the control group,the economic benefits of group 1,2,3 increased 2.28,4.80 and 4.09 yuan/d,respectively.In summary,dietary supplementation with 20 g/d of compound probiotics was the most effective method for milk performance,rumen fermentation parameters,nutrients apparent digestibility and economic benefit of Holstein dairy cows.
ABSTRACT
<p><b>OBJECTIVE</b>To observe the dynamic changes of membrane-bound proton-translocating ATPase (F-ATPase) in the development of dental caries, the expression of Streptococcus mutans F-ATPase under different pH concentrations and during the development of dental caries is analyzed.</p><p><b>METHODS</b>Streptococcus mutans cultured under different pH (pH4.0-7.0) concentrations and containing 5% glucose and no glucose containing BHI were collected. RNA was extracted. Subsequently, F-ATPase gene was detected using real-time polymerase chain reaction. Male Wistar rats were divided randomly into caries group and control group. The rats in the caries group were fed caries feed and 5% glucose water, whereas those of control group were fed normal feed. Total RNA was extracted from plaque samples, which were collected from rats' oral cavity every two weeks. F-ATPase gene was detected by real-time PCR. In the 11th week, the upper and lower jaw bone specimens of rats were taken, and molar caries damage assessed.</p><p><b>RESULTS</b>The expression of F-ATPase in the caries group was higher than that in the control group (P<0.05). In addition, the gene was expressed highest in pH5.0 and the lowest in pH4.0 (P<0.05). 2) The expression of F-ATPase progressively increased during the caries development in both groups; expression in the caries group was higher than that in control group (P<0.05).</p><p><b>CONCLUSION</b>Acid-resisting viru-lence factor F-ATPase is related closely with the incidence and development of dental caries.</p>
Subject(s)
Animals , Male , Rats , Adenosine Triphosphatases , Metabolism , Dental Caries , Metabolism , Microbiology , Dental Plaque , Microbiology , Protons , Random Allocation , Rats, Wistar , Real-Time Polymerase Chain Reaction , Streptococcus mutans , Genetics , Virulence FactorsABSTRACT
BACKGROUND:Scholars have carried out a lot of research on the main cariogenic microorganisms, but there is stil no comprehensive and systematic evaluation. OBJECTIVE:To perform an evidence-based medicine analysis on the six main cariogenic microorganisms. METHODS:Literature about microbial factors and age factors of caries disease etiology were retrieved, which were published from 2011 to 2014 in the Cochrane Library, EMBASE, PubMed, Wanfang, China Biology Medicine disc (CBMdisc), Chinese Journal Ful-text Database (CJFD) and VIP. Eligible studies were divided into two groups according to absorbance values: observation group (higher absorbance value) and control group (lower absorbance value). The probability of dental caries under the impact ofStreptococcus mutans, Lactobacilus microbe,Prevotela,Selenomonas,Capnocytophaga,Veilonelawere statisticaly analyzed using Meta-analysis. RESULTS AND CONCLUSION:Eight articles were included in result analysis. Odds ratio (OR) values refer to the statistical difference in the probability of dental caries under same pathogenic factors between two groups:OR > 1 indicates the incidence of dental caries is higher in the observation group than the control group, andOR < 1 suggests the incidence of dental caries is lower in the observation group than the control group. Meta-analysis results showStreptococcus mutans (OR=2.41),Lactobacilus microbe (OR=1.68),Prevotela (OR=2.75),Capnocytophaga (OR=1.98),Selenomonas(OR=2.38), indicating that the probability of dental caries is higher in the observation group than the control group, and these microorganisms are greatly correlated with the occurrence and development of dental caries;Veilonela (OR=0.45), indicating the probability of dental caries is lower in the observation group than the control group, andVeilonela has smal correlation with dental caries and only plays an important role in a certain development stage of dental caries.
ABSTRACT
Objective To investigate iodine nutritional status of pregnant women of Taizhou City,in order to provide a basis for assessment whether normal consumption of iodized salt is needed,to take complement iodine measure and to carry out monitoring of urinary iodine and thyroid function in pregnant women.Methods Early pregnancy (gestation ＜ 4 months),medium (4 to 7 months gestation) and late (pregnancy≥8 months) of each of more than 300 pregnant women were chosen from maternal and child health (hospital) and part of the township hospitals of Taizhou City (district) from September 2012 to December 2013.Based on the principle of informed consent,disposable urine and blood samples were collected,urine iodine and thyroid hormone [serum thyroid stimulating hormone (TSH),three free triiodothyronine (FT3) and free thyroid hormone (FT4)] were measured.Urinary iodine was measured using a urinary iodine rapid quantitative detection kit,thyroid function was measured by an electrochemical method.Use 95％ of the reference value range of thyroid function in pregnant women as the basis for evaluation of thyroid function in pregnant women.Results A total of 997 pregnant women were investigated,early stage,medium-term and late pregnancy women were 302,357 and 338,respectively.The median urinary iodine was 124,129,119 μg/L,pregnant women in late pregnancy had the lowest iodine urine.The median TSH of early stage,medium-term and late pregnancy women were 2.03,2.18,2.22 mU/L;the mean FT3 and FT4 of the three groups were (4.39 ± 0.66),(4.06 ± 0.61),(3.92 ± 0.61)pmol/L and (14.56 ± 2.56),(13.08 ± 2.03),(12.70 ± 2.38)pmol/L,respectively.The rate of abnormal thyroid function was 4.91％ (49/977),mainly in subhypothyroidism [48.98％ (24/49)].Conclusions Poor iodine-nutritional status exists in some pregnant women in Taizhou City.It is necessary to strengthen the propaganda and education of pregnant women on iodine supplement knowledge in the future,and carry out routine monitoring of urinary iodine and thyroid function in early pregnancy.
ABSTRACT
<p><b>OBJECTIVE</b>To explore the relationship between salivary proteome and dental caries and to promote the biomarker studies of dental caries susceptibility by comparing the salivary proteome of caries-active children and caries-free children with electrospray ionization ion-trap tandem mass spectrometry (ESI-MS/MS).</p><p><b>METHODS</b>Ten caries-active children and ten caries-free children were sampled. The salivary proteome of the two groups was studied, and the differential protein between the two groups was analyzed by ESI-MS/MS after sodium dodecyl sulfate polyacrylamide gel electrophoresis, filter-aided sample preparation, and liquid chromatography.</p><p><b>RESULTS</b>The concentration of salivary protein was higher in the caries-active group than in the caries-free group. The polypeptide counts of thecaries-active and caries-free groups were 602 and 481, which belonged to 286 and 227 proteins, respectively. The differential polypeptide count of the two groups was 361, and the differential protein count was 118. The detected proteins included matrix metalloproteinase-9 (MMP9), mucin-7 (MUC7), lactotransferrin (LTF), carbonic anhydrase 6 (CA6), azurocidin (AZU), and cold agglutinin.</p><p><b>CONCLUSION</b>The total salivary protein was higher in the caries-active group than in the caries-free group. The preliminary detection of differential proteins (MMP9, MUC7, LTF, CA6, AZU, and cold agglutinin) may lay some foundation for biomarker research of dental caries susceptibility.</p>
Subject(s)
Child , Humans , Carbonic Anhydrases , Dental Caries , Matrix Metalloproteinase 9 , Proteome , Saliva , Chemistry , Spectrometry, Mass, Electrospray Ionization , Tandem Mass SpectrometryABSTRACT
Objective To apply team based learning (TBL) in the teaching of dental endodontics for undergraduates in order to expand and deepen the TBL teaching in stomatology education.Methods TBL was used among 2008 grade oral undergraduates in Chongqing Medical University.Average test scores of 2008 grade undergraduates before and intra class were calculated and questionnaire was designed.At the same time,final examination scores between 2007 grade and 2008 grade under-graduates were compared.Results There was no significant correlation in average test scores before and intra class (r =0.027,P ＞ 0.05).Final examination scores were higher in 2008 grade than in 2007 grade.Based on the questionnaire survey,learning interests,sense of teamwork and classroom knowledge grasp of 2008 grade undergraduates were obviously elevated.Conclusions TBL teaching significantly improve students' learning effect and it can be promoted in stomotology education.
ABSTRACT
A 27-year-old woman who suffered from a 2-year history of systemic lupus erythematosus (SLE) presented with a 6-month history of nodules and ulcer on the right lower extremity. Direct microscopic examination of the pus showed branched and septate hyphae and spores in a chain-like arrangement.Histopathological examination revealed yellowish brown hyphae and spores. Dark green velvety colony grew on Sabouraud dextrose agar (SDA). Slide culture showed branched, septate hyphae and spine-like annellated conidiophores. The isolate was identified as Exophiala spinifera by DNA sequence analysis. The strain was unable to liquefy gelatin, could grow at 25 ℃ to 39 ℃, and was sensitive to itraconazole, amphotericin B and terbinafine. Animal test revealed that the infection induced by Exophiala spinifera in immunocompromised mice was more severe than that in normal controls. Based on the clinical features, histopathological, fungal culture and DNA sequencing results, the patient was diagnosed with systemic lupus erythematosus accompanied by subcutaneous phaeohyphomycosis caused by Exophiala spinifera.
ABSTRACT
Objective To report a case of cutaneous infection due to Trichosporon dermatis. Methods Lesional discharge and tissue were obtained and subjected to microscopic examination, fungal culture and histopathology, respectively. The fungal isolate was then identified with DNA sequence analysis, API 20C AUX system, gelatin liquefaction test, thermal tolerance test. Antifungal susceptibility test was also performed for the fungus. Results A 70-year-old male presented with a 9-month history of ulcerated swelling of the right medial malleolus after plant puncture. Direct microscopic examination of lesional discharge showed no fungal elements, but histopathological biopsy revealed hyphae and spores in the dermis. Yellowish white yeast-like colony grew on Sabouraud dextrose agar (SDA). Slide culture showed pseudohyphae, true hyphae, arthroconidia and blastoconidia. The isolate was identified as Candida humicola by API 20C AUX system, but as T. dermatis by DNA sequence analysis. The strain was unable to liquefy gelatin, could grow at 25 ℃ to 40 ℃, and was sensitive to amphotericin B, itraconazole, voriconazole and nystatin. The skin lesion completely subsided after 4-month treatment with oral itraconazole. Conclusions The isolate is identified as T. dermatis according to morphological features and DNA sequence, which is sensitive to itraconazole.
ABSTRACT
A 53-year-old male patient was admitted to the hospital on March 26, 2006. Ten years prior to the presentation, a small furuncle developed on the dorsum of his right hand, and subsided after 2-week treatment with erythromycin ointment; one month later, a broadbean-sized cutaneous ulcer developed on the dorsum of the same hand. After anti-infective treatment, the ulcer healed while the lesional skin thickened, and long-term topical treatment with compound dexamethasone acetate cream showed no obvious effect Dermatological examination revealed an irregular verrucous plaque measuring 2.5 cm × 4 cm with little exudation on the dorsum of the right hand. KOH preparation of the skin lesion revealed brown spores. Sabouraud's dextrose agar culture grew restricted, velvety and dark green colony, and microscopy revealed branched, globular conidiophores generated by cladospores. DNA sequencing showed that the isolate was different from Cladosporium sphaerospermum (AB100654) by 2 bases in the sequence of D1/D2 region of 26S rDNA, from Cladosporium sphaerospermum (AY625063) by 5 bases in the sequence of internal transcribed spacer 1 and 2 (ITS 1 and ITS 2), but fully consistent with Cladosporium sphaerospermum (AM 176719) in the sequence of ITS region. The isolate was identified as Cladosporium sphaerospermum. Hematoxylin-eosin stain of the lesional tissue revealed granulomatous changes,and PAS stain demonstrated brown spores. A diagnosis of phaeohyphomycosis was made. Antifungal susceptibility testing indicated that the isolate was highly sensitive to itraconazole. The lesion obviously subsided after treatment with oral itraconazole 0.2 g once daily for 8 weeks, but the patient was lost to follow up 2 months later.
ABSTRACT
Objective To report a case of superficial white onychomycosis caused by Nigrospora sphaerica. Methods Natl specimens were obtained from the patient and examined by direct microscopy, fungal culture and histopathology. Subsequently, the isolate was subjected to DNA sequencing analysis, gelatin liquefaction test, antifungal susceptibility test and nail-plate invasion test. Results A 21-year-old male presented with a 5-month history of whitening of the right hallux. Direct microscopy of nail scrapings showed spores, hyphae and lobiform conidiophores, and histopathology of decalcifying nail clippings revealed the presence of fungal elements including numerous spores and hyphae. A black woolly colony was formed in Sabouraud's dextrose agar (SDA). DNA sequencing analysis showed that the isolate was highly consis-tent with genus Nigrospora. Also, the isolate possessed the ability to liquefy gelatin and to invade normal nail plate. Antifungal susceptibility test showed that the isolate was highly susceptible to itraconazole, clotrimazole, amphotericin B and nystatin. The onychomycosis was cured after 5-month treatment with oral itra-conazole pulse therapy. Conclusions The isolate is identified as Nigrospora sphaerica by morphological features and DNA sequencing analysis. It is the first reported case of superficial white onychomycosis caused by N. Sphaerica in China, and it was effectively treated by itraeonazolc.