Effect of percutaneous transhepatic one-step biliary fistulation combined with rigid choledochoscopy in the treatment of benign hepatic bile duct strictures / 中华肝胆外科杂志

Objective:To investigate the therapeutic effect of percutaneous transhepatic one-step biliary fistulation (PTOBF) combined with rigid choledochoscopy in the treatment of benign hepatic bile duct strictures.Methods:The clinical data of 46 patients with hepatic ductal stenosis and stones undergoing PTOBF combined with rigid choledochoscopy at the First Hospital of Guangzhou Medical University between September 2016 and September 2022 were retrospectively analyzed, including 20 males and 26 females, aged (48.5±17.6) years. The location of stones and strictures, stone retrieval rate, postoperative complications, stricture release rate, stone recurrence rate, and reoperation rate were analyzed to access the safety and effectiveness of this procedure.Results:A total of 58 sites of stenoses were found in 46 patients, and PTOBF lithotripsy combined with rigid choledochoscopy were performed for 77 times. The early postoperative complication rate was 19.6% (9/46), and the stenosis release rate was 93.5% (43/46). The mean follow-up time was (28.1±19.6) months, the complete stone retrieval rate was 91.3% (42/46), the stone recurrence rate was 19.6% (9/46), the reoperation rate was 8.7% (4/46), and the long-term postoperative complication rate was 6.5% (3/46).Conclusion:PTOBF combined with rigid choledochoscopic stenoplasty for benign hepatic duct strictures is a safe and feasible procedure to reduce the stone recurrence and long-term postoperative complications.

A retrospective study on combined percutaneous transhepatic one-step biliary fistulation followed by two stages cholangioscopic treatment for type Ⅰ and Ⅱa hepatolithiasis after cholangiojejunostomy / 中华肝胆外科杂志

Objective:To study the treatment outcomes of combining percutaneous transhepatic one-step biliary fistulation (PTOBF) followed by two stages cholangioscopic treatment for type Ⅰ and Ⅱa hepatolithiasis which developed after Roux-en-Y cholangiojejunostomy, and in treatment of cholangiojejunostomy stenosis.Methods:The clinical data of 95 patients with type Ⅰ and Ⅱa hepatolithiasis which developed after Roux-en-Y cholangiojejunostomy and were treated at Shandong Second Provincial General Hospital from September 2016 to December 2020 were analyzed retrospectively. There were 36 males and 59 females, with the age of (51.2±15.3) years (range 14 to 75 years). These patients initially underwent PTOBF rigid choledochoscopy, followed by electronic choledochoscopy via the fistula tract after 6-8 weeks. The hepatolithiasis removal, complications and hepatolithiasis recurrence rates, and the cholangio-intestinal anastomotic stenosis rate and treatments were recorded. The follow-up was performed to analyse prognosis.Results:All 95 patients successfully underwent PTOBF rigid choledochoscopy and electronic choledochoscopy via the fistula tract. In 92 patients (96.8%), stones were completely removed. In 3 patients, small amounts of peripheral bile duct stones were left behind. Of 49 patients had cholangio-intestinal anastomotic strictures. On cholangioscopic examination, the strictures were caused by anastomotic knots in the suture line in 25 patients and cicatricial stenosis in 24 patients. After biliary balloon dilation and removal of anastomotic suture line knots, the strictures were relieved in 49 patients. There were 2 patients who developed biliary bleeding and 2 patients pleural effusion after PTOBF rigid choledochoscopy. Hepatolithiasis recurred in 4 patients in 6 to 36 months later.Conclusion:PTOBF followed by two stages cholangioscopic treatment were safe and effective in treatment of type Ⅰ and Ⅱa hepatolithiasis after Roux-en-Y cholangiojejunostomy. A high hepatolithiasis removal rate was obtained. Balloon dilation and removal of biliary intestinal anastomotic suture knots effectively relieved biliary intestinal anastomotic stenosis. The long-term results needs to be further determined.

One-step synthesis of site-specific antibody-drug conjugates by reprograming IgG glycoengineering with LacNAc-based substrates

Glycosite-specific antibody‒drug conjugatess (gsADCs), harnessing Asn297 N-glycan of IgG Fc as the conjugation site for drug payloads, usually require multi-step glycoengineering with two or more enzymes, which limits the substrate diversification and complicates the preparation process. Herein, we report a series of novel disaccharide-based substrates, which reprogram the IgG glycoengineering to one-step synthesis of gsADCs, catalyzed by an endo-N-acetylglucosaminidase (ENGase) of Endo-S2. IgG glycoengineering via ENGases usually has two steps: deglycosylation by wild-type (WT) ENGases and transglycosylation by mutated ENGases. But in the current method, we have found that disaccharide LacNAc oxazoline can be efficiently assembled onto IgG by WT Endo-S2 without hydrolysis of the product, which enables the one-step glycoengineering directly from native antibodies. Further studies on substrate specificity revealed that this approach has excellent tolerance on various modification of 6-Gal motif of LacNAc. Within 1 h, one-step synthesis of gsADC was achieved using the LacNAc-toxin substrates including structures free of bioorthogonal groups. These gsADCs demonstrated good homogeneity, buffer stability, in vitro and in vivo anti-tumor activity. This work presents a novel strategy using LacNAc-based substrates to reprogram the multi-step IgG glycoengineering to a one-step manner for highly efficient synthesis of gsADCs.

Effectiveness of different orthodontic adhesives and a universal bonding resin on the bond strength of brackets to different ceramics / Eficácia de diferentes adesivos ortodônticos e uma resina de ligação universal na resistência adesiva de bráquetes à diferentes cerâmicas

Objective: The aim of this study was to evaluate the effectiveness of different adhesives on the shear bond strength (SBS) of brackets bonded to different ceramic materials. Material and Methods: Fifty disk-shaped specimens were produced from lithium disilicate (IPS e.max CAD) and monolithic zirconia (Cercon) materials. Each specimen was polished with a three-step diamond polishing system. The polished ceramic surfaces were conditioned with universal bonding resin (Assure Plus) without pre-treatment, except for two specimens. Central brackets were bonded onto different ceramic specimens with different adhesives as follows: group 1: conventional adhesive onto the lithium disilicate; group 2: one-step adhesive onto the lithium disilicate; group 3: conventional adhesive onto the monolithic zirconia; group 4: one-step adhesive onto the monolithic zirconia. After thermal cycling, the specimens were subjected to the SBS test. The adhesive remnant index (ARI) scores were also recorded to evaluate bond failure type. Kruskal­Wallis and Mann-Whitney U tests were used for statistical analysis. Results: There were statistically significant differences among the SBS values (p˂0.05). The monolithic zirconia group with universal bonding resin and conventional orthodontic adhesive demonstrated the highest SBS value (6.34 MPa) and ARI scores. The lithium disilicate group showed the lowest SBS value (2.17 MPa) with the same protocol. No adhesive remained on the lithium disilicate specimens. Conclusion: One-step adhesive and universal bonding resin combination should not be considered as an alternative for lithium disilicate and monolithic zirconia restorations. Conventional adhesive and universal bonding resin application can be effective on non-pretreated ceramic surfaces during orthodontic bonding (AU)

Objetivo: o objetivo deste estudo foi avaliar a eficácia de diferentes adesivos na resistência ao cisalhamento (SBS) de bráquetes colados a diferentes materiais cerâmicos. Material e métodos: Cinquenta espécimes em forma de disco foram produzidos a partir de materiais de dissilicato de lítio (IPS e.max CAD) e zircônia monolítica (Cercon). Cada amostra foi polida com um sistema de polimento de diamante de três passos. As superfícies cerâmicas polidas foram condicionadas com resina de ligação universal (Assure Plus) sem pré-tratamento, exceto para dois corpos-de-prova. Bráquetes centrais foram colados em diferentes corpos de prova cerâmicos com diferentes adesivos da seguinte forma: grupo 1: adesivo convencional sobre dissilicato de lítio; grupo 2: adesivo de uma etapa sobre o dissilicato de lítio; grupo 3: adesivo convencional sobre zircônia monolítica; grupo 4: adesivo de uma etapa sobre a zircônia monolítica. Após a ciclagem térmica, os corpos-de-prova foram submetidos ao teste SBS. Os escores do índice de remanescente adesivo (ARI) também foram registrados para avaliar o tipo de falha de adesão. Os testes U de Kruskal-Wallis e Mann-Whitney foram usados para análise estatística. Resultados: Houve diferenças estatisticamente significativas entre os valores de SBS (p˂0,05). O grupo de zircônia monolítica com resina de colagem universal e adesivo ortodôntico convencional demonstrou o maior valor de SBS (6,34 MPa) e escores de ARI. O grupo de dissilicato de lítio apresentou o menor valor de SBS (2,17 MPa) com o mesmo protocolo. Nenhum adesivo permaneceu nas amostras de dissilicato de lítio. Conclusão: A combinação de adesivo de uma etapa e resina de ligação universal não deve ser considerada como uma alternativa para restaurações de dissilicato de lítio e zircônia monolítica. A aplicação de adesivo convencional e resina de colagem universal podem ser eficazes em superfícies de cerâmica não pré-tratadas durante a colagem ortodôntica (AU)

Three-dimensional visualization technology in one-stage establishment of a percutaneous transhepatic biliary tract in treatment of complex hepatolithiasis / 中华肝胆外科杂志

Objective:To study the use of percutaneous transhepatic one-step biliary fistulation based on three-dimensional visualization technology (3D-PTOBF) in the treatment of complex hepatolithiasis.Methods:A retrospective analysis was conducted on 116 patients with complex hepatolithiasis treated in the First Affiliated Hospital of Guangzhou Medical University from January 2016 to December 2019. There were 56 patients in the 3D-PTOBF group (treated by 3D-PTOBF), and 60 patients in the traditional PTOBF group (received traditional PTOBF approach). The stone clearance rate, postoperative complication rate, intraoperative blood loss, hospitalization time, number of cholangioscopic treatment procedures and stone recurrence rate were compared between the two groups.Results:When compared with the traditional PTOBF group, the 3D-PTOBF group had significantly less procedures (1.43±0.71 vs. 2.07±1.22, P<0.05), and shorter hospital stay (4.6±2.3 d vs. 6.1±2.9 d, P<0.05). There were no significant differences in the immediate stone clearance, final stone clearance, postoperative complications and stone recurrence rates between the two groups (all P>0.05). Conclusion:3D-PTOBF was safe and feasible to treat complex hepatolithiasis. When compared with PTOBF, it had the advantages of shorter operation time and decreased hospital stay.

Screening of gestational diabetes mellitus using one-step versus two-step method: a comparative study

Background: Gestational diabetes mellitus (GDM) is defined as carbohydrate intolerance of variable severity with onset or first recognition during the present pregnancy. It affects 7% of all pregnancies worldwide and in India it ranges from 6 to 9% in rural and 12 to 21% in urban area. The aim of this study was to compare the DIPSI criteria with the two-step method (Carpenter and Couston criteria.) and to study merits and demerits of one step and two step tests for GDM.Methods: A total 400 pregnant women of gestational age between 24-28 weeks attending antenatal clinic at this study tertiary care center were enrolled in this study. 200 pregnant women were enrolled in each of the study group (Group I OGTT and Group II DIPSI).Results: In Group I (OGTT) screening 47 (23.5%) were tested positive. In Group II cases, screening test results were found positive among 44 (22%). Out of 95 high-risk pregnant women 38 (40%) were positive for GDM by OGTT and 34 (35.78%) were positive by DIPSI. Out of 305 non high-risk pregnant women, 9 (2.95%) were positive for GDM by OGTT and 10 (3.27%) were positive by DIPSI.Conclusions: Present study concludes that DIPSI is the test which can predict GDM in population comparable to another test like OGTT. Also, India’s major population reside in rural areas, ANC are mostly conducted by ANM, therefore screening test should be easy to perform and interpret.

Biosynthesis of α-lipoic acid in Gluconobacter oxydans increases the production of vitamin C by one-step fermentation / 生物工程学报

In a one-step fermentation system of vitamin C production with Gluconobacter oxydans and Ketogulonicigenium vulgare, a functional module of α-lipoic acid biosynthesis was constructed in G. oxydans. The engineered G. oxydans was co-cultured with K. vulgare to enhance the growth and 2-keto-L-gulonic acid (2-KGA) production of K. vulgare. This one-step fermentation system alleviated the growth inhibition during the mono-culture of K. vulgare and strengthened the interaction between the two bacteria. Moreover, the yield of vitamin C precursor (2-KGA) increased to 73.34 g/L (the control group was 59.09 g/L), and the conversion of D-sorbitol to 2-KGA increased to 86.0%. This study provides a new idea for further optimizing the one-step fermentation system of vitamin C production.

Sentinel Lymph Node Analysis in Colorectal Cancer Patients Using One-Step Nucleic Acid Amplification in Combination With Fluorescence and Indocyanine Green

PURPOSE: Analysis of the sentinel lymph node (SLN) in colorectal cancer (CRC) patients was proposed for more accurate staging and tailored lymphadenectomy. The aim of this study was to assess the ability to predict lymph node (LN) involvement through analysis of the SLN with a one-step nucleic acid (OSNA) technique in combination with peritumoral injection of indocyanine green (ICG) and near-infrared (NIR) lymphangiography in CRC patients. METHODS: A total of 34 patients were enrolled. Overall, 51 LNs were analyzed with OSNA. LNs of 17 patients (50%) were examined simultaneously with hematoxylin and eosin (H&E) and OSNA. RESULTS: SLN analysis of 17 patients examined with H&E and OSNA revealed that OSNA had a higher sensitivity (1 vs. 0.55), higher negative predictive value (1 vs. 0.66) and higher accuracy (100% vs. 76.4%) in predicting LN involvement. Overall, OSNA showed a sensitivity of 0.69, specificity of 1, accuracy of 88.2%, and stage migration of 8.8%. Compared to those who were OSNA (−), OSNA (+) patients had a greater number of LN metastases (4.8 vs. 0.16, P = 0.04), higher G3 rate (44.4% vs. 4%, P = 0.01), more advanced stage of disease (stage III: 77.8% vs. 16%; P = 0.00) and were more rapidly subjected to adjuvant chemotherapy (39.1 days vs. 50.2 days, P = 0.01). CONCLUSION: SLN analysis with OSNA in combination with ICG-NIR lymphangiography is feasible and can detect LN involvement in CRC patients. Furthermore, it allows for more accurate staging reducing the delay between surgery and adjuvant chemotherapy.

New Culicoides Vector Species for BTV Transmission in Central and Central West of Anatolia

The aim of this study was to investigate the presence of BTV infection and possible vector species in different regions of Turkey. In the study, blood samples taken from 666 Akkaraman sheep were examined. 2000 Culicoides specimens were captured by light traps from the same provinces and 20 Culicoides spp. were identified. Blood sera samples were investigated by c-ELISA and SNT for detecting Abs to BTV. Sera samples were detected as positive 67 (10.06%) and 160 (24.02%) by SNT and ELISA, respectively. SN50 values of the 67 positive sera samples by SNT were detected between 1/2.38 and 1/200. All sheep blood samples and pools became Culicoides spp. samples were examined for BTV Ag presence by BTACE. Thirty six (5.40%) blood samples were detected as positive, but no from Culicoides pools. In the meantime, all sheep blood samples and Culicoides samples were directly investigated for BTV genome by one step RT-PCR. Fourteen (2.10%) blood samples and 7 (11.11%) Culicoides species were detected as positive. Also, the blood samples and the Culicoides samples were inoculated into Vero cell culture and passaged 5 times. Twenty nine (4.35%) blood samples cultured in Vero cell culture lines showed CPE but non CPE was observed in Culicoides samples. While 5 (17.24%) of 29 CPE positive isolates were identified as BTV by One Step RT-PCR. Total 26 samples (14 blood samples, 7 Culicoides samples and 5 supernatants) which detected BTV genome positive by One Step RT-PCR were serotyped. At the end of the study, while 23 of 26 samples were serotyped as BTV-9, two samples were serotyped as BTV-4. One sample (C. punctatus) from Culicoides was not serotyped as none of serotypes of BTV. In the present study, BTV was isolated for the first time from C. circumscriptus, C. kibunensis, and C. punctatus in Turkey.

Study on prescription and process of Fritillariae Thunbergii Bulbus formula granules based on characterization of powdery / 中草药

Objective: To study the prescription and process of Fritillariae Thunbergii Bulbus (FTB) formula granules. Methods: FTB was extracted by decocting method. FTB extract powder was prepared by spray drying method. Wet-extruding granulating, extruded-rolling granulating, and one-step granulating were adopted for preparing the formula granules. A comprehensive evaluation method was based on the powder fluidity parameters such as the rest angle, the final volume reduction degree (a), the filling velocity constant (b, k), and the yield of the particles, to optimize the optimum preparation process, prescription excipients and their proportion. An HPLC method was used to determine the contents of peimine and peiminine. The chromatographic column was the Agilent Zorbax Eclipse XDB-C18 column (150 mm × 4.6 mm, 5 μm), and the flow phase was Acetonitrile-water-diethylamine (70:30:0.03); The volume flow was 1.0 mL/min; The column temperature was 30 ℃. Results: The synthesis score of the FTB formula granules prepared by one-step granulating method was the highest. The best prescription was the extract powder-dextrin- 95% ethanol solution (100:100:160). The particle yield was 91.3%, the rest angle was 30.73°, the value of a was 0.109 1, the value of b was 0.025 5 and the value of k was 0.030 1, the fluidity of the granules was good and the yield was high. The content of peimine was 0.305% and the content of peiminine was 0.098% in the particles by HPLC. Conclusion: In this experiment, the formulation and process of FTB formula granules were designed to met the design requirements, which could be used in the production of the technology.

Optimization of One-step Pelletization Technology of Rouganbao Granules with Orthogonal Test Methodology / 中国中医药信息杂志

Objective To optimize the process conditions of one-step pelletization of Rouganbao Granules. Methods The factors influencing the pelletization of Rouganbao Granules were investigated by L9(34)orthogonal test, with the indexes of forming rate and fluidity. Results The spraying speed had the greatest effect on one-step pelletization, followed by atomization pressure and material temperature, and wind temperature had the smallest effect. At last, the best process parameters were relative density 1.15 (60 ℃), spray speed 55 mL/min, atomization pressure 0.25 MPa, wind temperature 75 ℃, material temperature 55 ℃, and critical relative humidity was 63%. Conclusion One-step pelletization technology can improve the preparation level and product quality, which can be used for the industrial production of Rouganbao Granules.

Green, Low-cost Ball Milling-Hydrothermal Synthesis ofGraphene Quantum Dots and Applications / 分析化学

A ball milling method which is green with simple-manipulation and low-cost was used to prepare graphene as precursor for graphene quantum dots (GQDs) synthesis.Subsequently, GQDs with good dispersibility, uniform size distribution, average diameter of (4.80 ± 0.20) nm and 1-3 layers were prepared by one-step hydrothermal method.The morphology, structure and optical properties of the GQDs were characterized by high-resolution transmission electron microscopy (HRTEM), atomic force microscope (AFM), Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), UV-Vis absorption and fluorescence spectroscopy.Furthermore, the GQDs were used in label-free and specific detection of ferric ion (Fe.3+) with broad linear ranges of 2.0×10.-6-7.0×10.-4 mol/L and low detection limit of 1.8 × 10.6 mol/L (S/N=3).The possible mechanism was also discussed and the application of GQDs for Fe.3+ detection in tap water was demonstrated.Finally, based on their low cytotoxicity and excellent biocompatibility, the as-prepared GQDs were successfully applied to efficient cell imaging.This work provides a new way for preparation of carbon-based nanomaterials and build a foundation for deepening applications of GQDs in bio-/chem-analysis, bioimaging, etc.

One-step dilation technique in minimally invasive percutaneous nephrolithotomy / 中国内镜杂志

Objective To evaluate the efficacy and safety of one-step dilation technique in minimally invasive percutaneous nephrolithotomy (MPCNL). Methods Clinical data of 2813 patients who underwent MPCNL by one-step dilation technique from February 2011 to March 2015 was retrospectively analyzed. Results 2813 patients were successfully underwent MPCNL by one-step dilation, including 2383 cases who were accessed by single tracts (84.71%) and 430 (15.29%) cases by multiple tracts. The mean operating time was (78.6 ± 41.1) min, the mean tract accessing time was (2.3 ± 0.8) min.The stone-free rate after one session operation was 78.59%. It improved to 91.50% one month after operation. During and after operation, 93 cases needed transfusion, 21 underwent selective renal artery embolization. Adjacent viscera damage: 9 cases with pleural lesions and 1 case with colon injury, 13 cases with urinary extravasation, perirenal hematoma in 15 cases, without liver and spleen injury. Septic shock in 2 cases, who was recovered after anti-infection treatment. Conclusion One-step dilation is safe and effective technique to establish tracts in MPCNL, which can reduce X-ray exposure and operation time, but does not increase the risk of bleeding.

Analysis of penicillamine using Cu-modified graphene quantum dots synthesized from uric acid as single precursor / 药物分析学报

A simple methodology was developed to quantify penicillamine (PA) in pharmaceutical samples, using the selective interaction of the drug with Cu-modified graphene quantum dots (Cu-GQDs). The proposed strategy combines the advantages of carbon dots (over other nanoparticles) with the high affinity of PA for the proposed Cu-GQDs, resulting in a significant and selective quenching effect. Under the optimum conditions for the interaction, a linear response (in the 0.10–7.50 μmol/L PA concentration range) was observed. The highly fluorescent GQDs used were synthesized using uric acid as single precursor and then characterized by high resolution transmission electron microscopy, Raman spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, fluorescence, and absorption spectroscopy. The proposed methodology could also be extended to other compounds, further expanding the applicability of GQDs.

One-step dilation technique in minimally invasive percutaneous nephrolithotomy / 中国内镜杂志

Objective To evaluate the efficacy and safety of one-step dilation technique in minimally invasive percutaneous nephrolithotomy (MPCNL). Methods Clinical data of 2813 patients who underwent MPCNL by one-step dilation technique from February 2011 to March 2015 was retrospectively analyzed. Results 2813 patients were successfully underwent MPCNL by one-step dilation, including 2383 cases who were accessed by single tracts (84.71%) and 430 (15.29%) cases by multiple tracts. The mean operating time was (78.6 ± 41.1) min, the mean tract accessing time was (2.3 ± 0.8) min.The stone-free rate after one session operation was 78.59%. It improved to 91.50% one month after operation. During and after operation, 93 cases needed transfusion, 21 underwent selective renal artery embolization. Adjacent viscera damage: 9 cases with pleural lesions and 1 case with colon injury, 13 cases with urinary extravasation, perirenal hematoma in 15 cases, without liver and spleen injury. Septic shock in 2 cases, who was recovered after anti-infection treatment. Conclusion One-step dilation is safe and effective technique to establish tracts in MPCNL, which can reduce X-ray exposure and operation time, but does not increase the risk of bleeding.

Analysis of penicillamine using Cu-modified graphene quantum dots synthesized from uric acid as single precursor / 药物分析学报

A simple methodology was developed to quantify penicillamine (PA) in pharmaceutical samples, using the selective interaction of the drug with Cu-modified graphene quantum dots (Cu-GQDs). The proposed strategy combines the advantages of carbon dots (over other nanoparticles) with the high affinity of PA for the proposed Cu-GQDs, resulting in a significant and selective quenching effect. Under the optimum conditions for the interaction, a linear response (in the 0.10–7.50 μmol/L PA concentration range) was observed. The highly fluorescent GQDs used were synthesized using uric acid as single precursor and then characterized by high resolution transmission electron microscopy, Raman spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, fluorescence, and absorption spectroscopy. The proposed methodology could also be extended to other compounds, further expanding the applicability of GQDs.

One Step Operation of the Persistent Pupillary Membrane Removal and Cataract Operation: A Case Report

PURPOSE: We report a case of one-step operation of persistent pupillary membrane removal, phacoemulsification, and posterior chamber lens implantation that was performed in a patient with persistent pupillary membrane and brunescent cataracts in both eyes. CASE SUMMARY: A 64-year-old male with no baseline disease visited our clinic with bilateral decreased visual acuity. His best corrected visual acuity at first visit was 0.1 in the right eye and 0.2 in the left eye. On anterior segment examination, both bilateral persistent pupillary membranes and brunescent cataracts were observed. First, we surgically removed the left pupillary membrane and performed phacoemulsification using posterior chamber lens implantation via one-stage operation. After one week, the same operation was performed for the right eye using the same method. At 6-months postoperative, his best corrected visual acuity was 0.2 in the right eye and 0.8 in the left eye. No complications such as anterior segment inflammation, uveitis, or intraocular pressure elevation were observed during the follow-up period. CONCLUSIONS: We report a case of one-step operation of persistent pupillary membrane removal and cataract operation, which can improve visual acuity without any complications.

One Step Extraction of Mycoplasma Pneumoniae Genomic DNA in Application Research of the Visuable LAMP System / 现代检验医学杂志

Objective The modified rapid one-step extraction (ROSE) method was employed in the Mycoplasma pneumoniae (Mp)genomic DNA extraction,in order to establish an easy DNA extraction method,which is used in Mp Loop Mediated Isothermal Amplification(LAMP) and suitable for high throughput without any expensive instruments.Methods In line with the character of Mp,a one-step Mp acid extraction method was established on the basis of ROSE.The studies had been carried out on comparing the modified ROSE DNA extraction with the classical methods of Boiling,CTAB,Alkali solution and reagent kit in extraction quality,quantity,reagent kinds,operation steps and extraction time.The extracted DNA of the Mp culture solution by these five methods had been used in LAMP to test the extraction efficiency.20 cases of Throat swab specimens from children with mycoplasma pneumonia were selected from January to March of 2016,and the throat swab specimens were extracted by the improved ROSE method then applied to LAMP detection system.Results Purity,comparison of ROSE method and other four methods were P＜0.05,the difference was statistically significant and ROSE was slightly better than the alkaline lysis method and boiling method,but lower than the CTAB method and kit method;Yield,results were compared with ROSE method and other four methods:there was no significant difference in q=0.95 (P＞0.05) compared with ROSE method and boiling method.The ROSE method was significantly different from the other three methods (P＜0.05),so the yield of ROSE method was higher than that of alkaline lysis method,CTAB method and kit method.Instrument and equipment,extracting procedure,extracting time and quantity ROSE,Boiling and Alkali solution were obvious ly better than CTAB and reagent kit.The extracted DNA by these five methods could be used in LAMP reaction,and the results showed no significant difference.Conclusion This study established an original one-step DNA genomic extraction method of Mp,which had been successfully applied in the Mp LAMP withoutany expensive instruments.Offer a groundbreaking method detecting Mp for the hospitals at all levels including the primary medical organizations.

Canine distemper virus detection by different methods of One-Step RT-qPCR / Detecção do vírus da cinomose canina por diferentes métodos de One-Step RT-qPCR

ABSTRACT: Three commercial kits of One-Step RT-qPCR were evaluated for the molecular diagnosis of Canine Distemper Virus. Using the kit that showed better performance, two systems of Real-time RT-PCR (RT-qPCR) assays were tested and compared for analytical sensitivity to Canine Distemper Virus RNA detection: a One-Step RT-qPCR (system A) and a One-Step RT-qPCR combined with NESTED-qPCR (system B). Limits of detection for both systems were determined using a serial dilution of Canine Distemper Virus synthetic RNA or a positive urine sample. In addition, the same urine sample was tested using samples with prior centrifugation or ultracentrifugation. Commercial kits of One-Step RT-qPCR assays detected canine distemper virus RNA in 10 (100%) urine samples from symptomatic animals tested. The One-Step RT-qPCR kit that showed better results was used to evaluate the analytical sensitivity of the A and B systems. Limit of detection using synthetic RNA for the system A was 11 RNA copies µL-1 and 110 RNA copies µl-1 for first round System B. The second round of the NESTED-qPCR for System B had a limit of detection of 11 copies µl-1. Relationship between Ct values and RNA concentration was linear. The RNA extracted from the urine dilutions was detected in dilutions of 10-3 and10-2 by System A and B respectively. Urine centrifugation increased the analytical sensitivity of the test and proved to be useful for routine diagnostics. The One-Step RT-qPCR is a fast, sensitive and specific method for canine distemper routine diagnosis and research projects that require sensitive and quantitative methodology.

RESUMO: Três kits comerciais de One-Step RT-qPCR foram avaliados para o diagnóstico molecular do Vírus da Cinomose Canina.Utilizando o kit que apresentou melhor desempenho, dois sistemas de RT-PCR em tempo real (RT-qPCR) foram comparados quanto à sensibilidade analítica na detecção do RNA do Vírus da Cinomose Canina:One-Step RT-qPCR (Sistema A) e One-Step RT-qPCR seguido da NESTED-qPCR (Sistema B).Os limites de detecção dos dois sistemas foram determinados utilizando diluição seriada de RNA sintético do Vírus da Cinomose Canina ou de uma amostra de urina positiva. Adicionalmente, uma amostra de urina foi avaliada com centrifugação ou ultracentrifugação prévia. Os kits comerciais de One-Step RT-qPCR amplificaram o RNA do vírus da cinomose canina em 10 (100%) amostras de urinas de animais sintomáticos. O kit de One-Step RT-qPCR que apresentou melhor resultado foi utilizado para avaliar a sensibilidade analítica dos sistemas A e B. Na reação da curva padrão com RNA sintético, o limite de detecção do sistema A foi de 11 cópias de RNA µL-1. No sistema B foi de 110 cópias de RNA µL-1 na One-Step RT-qPCR e 11 cópias de RNA µL-1 na NESTED-qPCR. A relação entre os valores de Ct e concentração de RNA foi linear. O RNA extraído das diluições da urina foi detectado nas diluições de 10-3 e10-2 pelos sistemas A e B, respectivamente. A centrifugação prévia da urina aumentou a sensibilidade analítica da análise e mostrou ser importante para a rotina diagnóstica. A reação de One-Step RT-PCR é um método rápido, sensível, específico e aplicável na rotina de diagnóstico molecular da cinomose e em projeto de pesquisa que requer metodologia quantitativa e sensível.