A rapid and sensitive method of loop-mediated isothermal amplification(LAMP) was established to detect Pseudomonas aeruginosa(P.aeruginosa).Three pairs of LAMP primers(inner,outer and ring primers) were designed according to the gbca gene of P.aeruginosa.Since adding hydroxy naphthol blue(HNB) to the reaction system, a positive reaction was indicated by a colorchange before and after the reaction,and was verified by agarose gellectrophoresis.Both LAMP and PCR were applied to detect clinical specimens, the sensitivity and specificity of the detection method were evaluated,and were compared with those of conventional PCR.A LAMP method for detecting P.aeruginosa was successfully established.The LAMP method showed specificity for P.aeruginosa without other bacteria amplification.The established LAMP method in this study enables rapid,sensitive and specific detection of P.aeruginosa,and can be applied for grass roots and small scale laboratories as well as field surveillance.

The paper is to report the observation of pharmacokinetic changes of the characteristic ingredients in the combinations of different-dose herbs of Shaoyao-Gancao decoction. After the establishment of HPLC analytical method of plasma effective constituents, rats were orally administered with different-dose herbs of Shaoyao-Gancao decoction. Blood samples at different times after administering these decoctions were collected, and then were analyzed by HPLC fingerprints technology. Pharmacokinetic parameters of characteristic peaks were analyzed by SPSS 15.0 software and DAS 2.0. At last, we looked for the correlation of those pharmacokinetic parameters and the dosage of Gancao. The best dose of Shaoyao-Gancao decoction was at the ratio of 4 to 4, which was consistent with the dose commonly used in ancient times. The absorption of characteristic peaks from Shaoyao-Gancao decoction was related with the dosage of Gancao, and there existed interaction between each characteristic ingredients. There existed the right dose-ratio of Shaoyao and Gancao to get the best effect. The absorptions of effective constitutents were mutual waxing and waning in order to increase biological effects together. It's demonstrated the compatibility connotation at a right dose-ratio of Shaoyao-Gancao decoction through the angle of pharmacokinetics.

The pharmacokinetics of a long-circulating PEGylated Radix Ophiopogonis polysaccharide (ROP) was investigated in rats following i.v. or s.c. administration at three dose levels (9, 20, 50 mg x kg(-1)). A moderate coupling reaction between the hydroxyl-activated ROP and the amino-terminated mPEG was chosen to produce PEGylate ROP. The grafting degree of the prepared conjugate was 1.03, and the molecular mass of mPEG used was 20 kDa. High-performance gel permeation chromatorgraphy with fluorescein isothiocyanate prelabeling was established to determine levels of the conjugate in plasma. The results showed that the elimination half-life of the conjugate following s.c. administration was basically identical to that after iv administration. An accurate linear correlation was observed between administration doses and areas under the curve of plasma conjugate level vs. time profile, regardless of the administration route. The absolute bioavailability of the conjugate following sc administration was approximately 56%, and the mean in vivo residence time was 52.1 h, increased 2.4 times compared to those of iv administration. In general, linear pharmacokinetics was observed for the conjugate within the dose range studied, and sc should be a promising administration route for the conjugate.

Objective To study on FBD (composed with Poria cocos, Atractylodes macrocephala, and Angelica sinensis) used to improve learning and memory function of animals. Methods The mice were treated by ig FBD with the doses of 35, 70 and 140 mg/kg for one week continuously. Effect of FBD on dysmnesia of acquired learning of mice induced by scopolamine, dysmnesia of memory retention of mice induced by NaNO2, and dysmnesia of reappearance of memory of mice induced by 45% ethanol were studied. Improvement for the dysmnesia of memory due to chronic blood deficiency in brain of rats induced by ligating the carotid arteries of two sides were observed. Results FBD can improve the dysmnesia of mice induced by scopolamine, NaNO2, and 45% ethanol significantly, also dysmnesia due to chronic blood deficiency in brain of rats induced by ligating the carotid arteries. Conclusion FBD has the function to improve the learning and memory function of animals.

Objective To assess the feasibility of characterizing pleural fluid on the basis of spectral imaging features utilizing spectral CT imaging. Methods Gemstone spectral imaging(GSI) was used to examine 20 pleural fluids filled tubes (11 exudates and 9 transudates ) following diagnostic thoracentesis. Effusions were classified as transudates or exudates using laboratory markers based on Light criteria. CT values on 140 kVp QC image were compared between two groups. Using GSI viewer, various CT spectral imaging parameters (CT values on different energy level, effective-Z, iodine-water concentration,calcium-water concentration and calcium-fat concentration ) were calculated and compared between two groups. The difference of these spectral characteristic parameters was evaluated statistically by independent-samples t test. Results According to Light criteria, the mean CT value on QC image of exudates [ ( 19. 56 ±4. 10) HU ] was higher than that of transudates [ ( 13.44 ±3.46) HU] (t =3.002,P =0. 010).Difference of CT value was found more obvious in the lower keV. On 40 keV images, the difference of CT value of two groups was the largest, the mean value of exudates [ (47.49 ± 14. 60) HU ] was significantly higher than that of transudates[ ( 19. 76 ± 6. 85) HU ] ( t = 5.520, P = 0. 000). While On 140 keV, the mean CT value were (9.76 ±4. 16)and (6.22 ±3. 17) HU and the difference of the two group has no statistically significant difference (t =2. 107,P =0. 050). The mean slope rates of exudates (0.51 ± 0.23)was significantly larger than that of transudates (0. 18 ± 0. 08 ) ( t= 4. 287, P = 0. 001 ). The effective-Z (7. 89 ± 0. 16), iodine-water concentration [ (5. 74 ± 1.28 ) g/L], calcium-water concentration[ (7. 89 ±1.78) g/L] and calcium-fat concentration [ (25.95 ± 1.74) g/L] of exudates were significantly higher than those of transudates [ 7.67 ± 0. 07, ( 1.70 ± 0. 95 ) g/L, (2. 53 ± 1.37 ) g/L, ( 20. 82 ± 1.40 ) g/L ] ( t = 4. 080,6. 998,6. 546,6. 301 ,P ＜ 0. 05 ). Conclusions The spectral curve and spectral imaging parameters of exudates is found to be different from transudates. The low energy spectral imaging plays an important role in the characterization of pleural fluid. Gemstone spectral CT imaging provides a new multiparameter method to differentiate transudates and exudates.

With the development of various diagnostic imaging techniques and minimally invasive treatment tech-niques, the diagnosis and treatment of benign biliary stenosis caused by chronic pancreatitis develop to non-invasive and minimally invasive. The newly developed imaging technology has the characteristics of clear, noninvasive,wide application and high diagnostic rate. In recent years the progress of minimally invasive operation mode compared to the original open operation has less trauma, less blood loss, lower incidence of postoperative complications and faster post-operative recovery and other advantages. The latest da Vinci robotic surgical system has been applied in China and has gradually solved the limitations of laparoscopic surgery,such as the field of vision, the operation is not flexi ble. Ad-vances in diagnosis and treatment of benign biliary stenosis caused by chronic pancreatitis are reviewed for this article for reference.

OBJECTIVE: To provide genetic testing and prenatal diagnosis for a woman with Sheldon-Hall syndrome. METHODS: The woman was subjected to targeted capture and next-generation sequencing for variant of genes associated with skeletal disorders. And the result was verified in her parents and fetus. RESULTS: The woman was found to harbor a c.188G>A variant of the TNNT3 gene, which was also found in her affected mother and the fetus. Her grandmother and grandmother's brother had similar manifestations, which was in line with an autosomal dominant inheritance. The same variant was not found in her father. CONCLUSION The c.188G>A variant of the TNNT3 gene probably underlay the distal joint contracture in this pedigree, based on which prenatal diagnosis was attained.

OBJECTIVE: To detect pathogenic variant in a child featuring Usher syndrome type II. METHODS: Peripheral blood samples of the child and his parents were collected for the analysis of variants of hearing impairment-related genes. The findings were verified in 100 individuals with normal hearing. RESULTS: The child was found to harbor compound heterozygous variants of the USH2A gene, namely c.8224-1G>C in intron 41 and c.5678C>G(p.Ser1893X) in exon 28, which were inherited respectively from his mother and father. Based on the American College of Medical Genetics and Genomics standards and guidelines, both c.8224-1G>C and c.5678C>G(p.Ser1893X) variants of USH2A gene were predicted to be pathogenic(PVS1+PM2+PM3). CONCLUSION The compound heterozygous variants c.8224-1G>C and c.5678C>G of the USH2A gene probably underlay the disease in this child. Above finding has enriched the spectrum of USH2A gene variants.
Child ; Exons ; Extracellular Matrix Proteins/genetics* ; Family ; Humans ; Introns ; United States ; Usher Syndromes/genetics*

Objective:To investigate the clinical value of targeted sequencing panel in the detection of genetic variation in neonates in neonatal intensive care unit (NICU).Methods:All neonates (≤28 d of age) admitted in the NICU (case group) and 200 full-term healthy neonates born with no obvious phenotypic abnormalities of Huzhou Maternity and Child Health Care Hospital were enrolled in this prospective study from November 2022 to January 2023. Based on a list of preventable and treatable rare diseases as well as newly screened diseases in China, a targeted sequencing panel suitable for Chinese newborns was designed to target the pathogenic genes and mutation sites associated with 601 genes and 542 diseases. Dried blood spot specimens were prepared and analyzed by the targeted sequencing panel. Pathogenic sites detected by the panel sequencing were verified using Sanger sequencing. The genetic testing results were analyzed according to the clinical features of the neonates. According to the number of primary clinical diagnosis index (including premature infants, neonatal hyperbilirubinemia, hemorrhagic diseases, neonatal infections, ventricular septal defect/patent ductus arteriosus, and others), these patients were divided into four groups with 1, 2, 3, and ≥4 diagnosis index, respectively. Chi-square test and linear correlation Chi-square test were used for statistical analysis. Results:There were 173 patients in the case group and 30.6% (53/173) of them carried pathogenic variants, including 52 positive for pathogenic genes and one with chromosome copy number variant. The positive rate of pathogenic genes was significantly higher in the case group than in the control group [30.1% (52/173) vs. 15.0% (30/200), χ 2=12.26, P<0.001]. Fourteen pathogenic genes were detected in the case group, including FLG, UGT1A1, G6PD, MYH7, AR, ABCC2, ACADS, CYP21A2, GJB2, MEFV, PAH, PKHD1, SCN4A, and HBA. In the case group, the detection rate of pathogenic variants in jaundiced neonates was higher than that in non-jaundiced neonates [35.2% (44/125) vs. 18.8% (9/48), χ 2=4.42, P=0.036]. However, there were no statistically significant differences in the detection rates of pathogenic variants between male and female infants, infants born to mothers of advanced maternal age or not, infants born to mothers with or without gestational diabetes mellitus, premature and term infants, or infants with or without hemorrhagic disorders, neonatal infections, or ventricular septal defects/patent ductus arteriosus in the case group (all P>0.05). The detection rate of pathogenic variants showed a linear increase in infants with 1, 2, 3, and ≥4 diagnosis index [21.1% (8/38), 25.4% (15/59), 38.2% (13/34), and 40.5% (17/42); linear correlation χ 2=4.84, P=0.028]. In the case group, seven genes with a high detection rate of genetic variation (including positive pathogenic genes and carriers) were UGT1A1 [had the highest detection rate, 24.9% (43/173)], GJB2, FLG, DUOX2, ABCA4, G6PD, and MUT. Seven loci with higher mutation frequency were c.211G>A(p.Gly71Arg), c.1091C>T(p.Pro364Leu), c.-41_-40dupTA, and c.686C>A(p.Pro229Gln) in the UGT1A1 gene, c.109G>A(p.Val37Ile) in the GJB2 gene, and c.12064A>T(p.Lys4022Ter) and c.3321del(p.Gly1109GlufsTer13) in the FLG gene. Conclusion:This panel sequencing can provide effective genetic testing for neonates in NICU, especially in children with complex clinical diagnosis.