Objective To establish and evaluate a rapid detection method for SARS-CoV-2 based on reverse transcriptase-recombinase aided amplification(RT-RAA)combined with the clustered regularly interspaced short palindromic repeats(CRISPR)/Cas12a system.Methods RT-RAA primers and CRISPR-derived RNA(crRNA)were designed based on the nucleocapsid(N)gene of SARS-CoV-2 from NCBI database.The detection system was optimized with magnesium acetate(MgAc)concentration,RT-RAA reaction tempera-ture and time and LbCas12a reaction temperature.The sensitivity and specificity of the method were evaluated using recombinant plas-mids(100-106 copies/μL)and other respiratory pathogens.The RT-RAA-CRISPR/Cas12a method was compared with RT-PCR by tes-ting 70 clinical samples in parallel.Results The optimized RT-RAA-CRISPR/Cas12a assay could detect SARS-CoV-2 within 50 min at 37 ℃.The limit of detection was 10 copies/μL for the fluorescence-based method and 1×102 copies/μL for the lateral flow assay.The method specifically detected SARS-CoV-2 without cross-reactivity to other respiratory pathogens.The results of testing 70 clinical samples using RT-RAA-CRISPR/Cas12a showed agreement of 100％with those of RT-PCR.Conclusion The established RT-RAA-CRISPR/Cas12a assay for SARS-CoV-2 detection is rapid,cost-effective,highly sensitive and specific.It can be performed by less experienced personnel and no expensive equipment is required,thus it may provide a new approach for rapid clinical diagnosis and large-scale on-site screening of SARS-CoV-2.

Objective:To understand the breastfeeding situation in the neonatal intensive care units (NICUs) in Qianxinan Prefecture, Guizhou Province, and to assess the knowledge, attitudes, and practices of doctors and nurses regarding breastfeeding, aiming to provide foundational data for improving breastfeeding quality.Methods:A questionnaire was developed to survey the knowledge, attitudes, and practices related to breastfeeding in NICUs. The questionnaire was divided into three dimensions: knowledge (seven items, total score of 7), attitudes (nine items, total score of 45), and practices (seven items, total score of 35). Lower scores indicated weaker recognition of breastfeeding. Additionally, five items were included to identify the most influential factors affecting breastfeeding. From November 25 to November 30, 2023, a survey was conducted among doctors and nurses with professional qualifications who had worked in the neonatal departments of nine hospitals in Qianxinan Prefecture for at least one year. Independent sample t-tests and Chi-square tests were used to compare the scores of doctors and nurses from different levels of hospitals and within the same level of hospitals across the three dimensions. Results:(1) Among the nine hospitals, three were tertiary grade A hospitals (referred to as "tertiary hospitals"), with 95.6% (43/45) of the doctors and 96.5% (110/114) of the nurses participating in the survey. Six were secondary grade A hospitals (referred to as "secondary hospitals"), with 95.0% (38/40) of the doctors and 97.6% (83/85) of the nurses participating. (2) All nine hospitals were baby-friendly hospitals and all had breastfeeding promotional materials. Six hospitals had NICUs that promoted breastfeeding, with an average NICU breastfeeding rate of 25.8% across the prefecture between year 2021 to 2023. (3) The proportion of doctors who had received breastfeeding training was higher than that of nurses within the same level of hospitals [tertiary hospitals: 69.8% (30/43) vs. 40.0% (44/110), χ 2=10.97, P=0.001; secondary hospitals: 47.4% (18/38) vs. 24.1% (20/83), χ 2=6.55, P=0.010], although the overall training rates were low. (4) In tertiary hospitals, doctors scored higher than nurses in the attitude dimension [(35.35±4.75) vs. (33.18±5.60) scores, t=－2.03, P=0.044] and also in the practice dimension [(26.98±3.00) vs. (25.60±3.75) scores, t=－2.17, P=0.032]. In secondary hospitals, the total knowledge dimension score of doctors was higher than that of nurses [(4.92±1.44) vs. (4.20±1.45) scores, t=－2.52, P=0.013]. In tertiary hospitals, the total scores for attitude and practice dimensions of doctors were higher than those of doctors in secondary hospitals, and the total scores for knowledge, attitude, and practice dimensions of nurses were higher than those of nurses in secondary hospitals (all P<0.05). (5) In the knowledge dimension, the lowest scoring item of doctors in the tertiary hospitals was "Breastfeeding is possible for maternal hepatitis B newborns after receiving vaccines and immunoglobulin"; the lowest scoring item of nurses in the tertiary hospital, and doctors and nurses in the secondary hospitals was "The duration of breastfeeding has a greater impact on neonatal outcomes". In the attitude dimension, the lowest scoring item for doctors and nurses in both tertiary and secondary hospitals was "You think the breastfeeding process is more troublesome than feeding preterm formula". In the practice dimension, the lowest scoring item of the doctors and nurses in the tertiary hospitals was "Your hospital had enough breastfeeding knowledge training", while for the doctors and nurses in the secondary hospitals were "You have more opportunities to participate in various breastfeeding-related training" and "Breast feeding should be started as soon as possible when the infant is stable after active treatment", respectively. (6) The most influential factors affecting breastfeeding were: lack of cooperation from parents (50.0%, 137/274), relative insufficient human resources for doctors and nurses (21.9%, 60/274), and the absence or poor implementation of breastfeeding management policies (18.3%, 50/274), etc. Conclusions:The breastfeeding rate in NICU of county-level hospitals is relatively low, and medical staff, especially nurses, have insufficient knowledge about breastfeeding. It is necessary to strengthen various breastfeeding training for medical staff to enhance their understanding of NICU breastfeeding.

To explore the pharmacogenomic markers that affect the platinum-based chemotherapy response in non-small-cell lung carcinoma (NSCLC), we performed a two-cohort of genome-wide association studies (GWAS), including 34 for WES-based and 433 for microarray-based analyses, as well as two independent validation cohorts. After integrating the results of two studies, the genetic variations related to the platinum-based chemotherapy response were further determined by fine-mapping in 838 samples, and their potential functional impact were investigated by eQTL analysis and in vitro cell experiments. We found that a total of 68 variations were significant at P < 1 × 10^-3 in cohort 1 discovery stage, of which 3 SNPs were verified in 262 independent samples. A total of 541 SNPs were significant at P < 1 × 10^-4 in cohort 2 discovery stage, of which 8 SNPs were verified in 347 independent samples. Comparing the validated SNPs in two GWAS, ADCY1 gene was verified in both independent studies. The results of fine-mapping showed that the G allele carriers of ADCY1 rs2280496 and C allele carriers of rs189178649 were more likely to be resistant to platinum-based chemotherapy. In conclusion, our study found that rs2280496 and rs189178649 in ADCY1 gene were associated the sensitivity of platinum-based chemotherapy in NSCLC patients.

Objective:Intensive physicians have relatively insufficient knowledge and experience in treating patients with decompensated schistosomiasis cirrhosis (DSC) admitted in intensive care unit (ICU), but are relatively familiar with patients with decompensated alcoholic cirrhosis (DAC). For this purpose, the clinical characteristics and prognosis of these patients were compared and analyzed.Methods:A retrospective analysis was performed from January 2013 to May 2019 in our hospital and Quzhou People’s hospital. The demographic data, laboratory examination, liver function, Child-Pugh classification, complications of cirrhosis, ultrasonic imaging gastroscopy manifestations were recorded and analyzed. In addition, the treatments and prognosis were also compared.Results:A total of 30 patients (12 males and 18 females) with DSC (aged 57-88) and 31 patients with DAC (aged 41-75) were collected. Compared with patients with DAC, DSC patients were more likely to have coronary heart disease, lower proportion of hyponatremia and lower need of ventilator support. Although the incidences of jaundice and hepatic encephalopathy were significantly reduced ( P<0.05), but parameters of liver function and coagulation were no significant differences in both groups ( P>0.05). B-mode ultrasound of liver in patients with DSC displayed more proportion of patchy and diffuse echo changes and liver volume reduction ( P<0.05), whereas the manifestations of gastroscope in both groups were similar. No significant difference in main treatment measures like uses of somatostatin and three-chamber and two-capsule tube was observed. After treatment, the stop time of gastrointestinal bleeding was similar between groups of DAC and DSC [1.25 (0.5-4.125) days vs. 1.75 (1-2.375) days] ( P>0.05). In addition, the length of ICU stay in DAC group was similar to DSC group [(4.96±3.58) days vs. (3.82±1.99) days], so did the 28-day mortality [14.29% (2/14) vs. 18.18% (2/11)] (both P>0.05). Conclusions:In genenal, patients with decompensated schistosomiasis cirrhosis have the similar clinical characteristics, major biochemical indicators and accessory examination results like ultrasound and gastroscopic examinations to patients with decompensated alcoholic cirrhosis. After timely treatments, both of these patients could achieve a good prognosis.

OBJECTIVE： To investigate the changes of extraction rates of forsythiaside A and forsythin in Forsythia suspensa compatible with other medicinal material of Menshi huwei formula before and after decoction. METHODS： HPLC method was used to determine the extraction amounts and to calculate the extraction rates of forsythiaside A and forsythin in F. suspensa （5 g×7 doses）， F. suspensa （5 g×7 doses） compatible with Pinelliae rhizoma praeparatum cum zingibere et alumine （PRZA）， Menshi huwei formula [including 6 ingredients as F. suspense （5 g×7 doses）， PRZA] after decocted with water. The determination was performed on Diamonsil C18 column with mobile phase consisted of acetonitrile-0.2％ formic acid solution （gradient elution）. The detection wavelength was set at 278 nm， and the column temperature was 30 ℃. The flow rate was 1.0 mL/min. RESULTS： The linear range of forsythiaside A and forsythin were 0.61-6.1， 0.246-2.46 μg （r＝0.999 7， 0.999 9）， respectively； RSDs of precision， stability （within 20 h） and reproducibility tests were all lower than 2％ （n＝6）. Average recovery rates were 96.10％-99.37％ （RSD≤2.36％，n＝6） respectively. In F. suspensa， extraction rates of forsythiaside A and forsythin were 96.90％ and 66.67％. In F. suspensa compatible with PRZA， extraction rates of them were 101.61％ and 54.55％. In Menshi huwei formula， extraction rates of them were 98.39％ and 84.85％. CONCLUSIONS： After F. suspensa is compatible with PRZA， the extraction rates of forsythiaside A is increased while forsythin is decreased. After compatible with other medicinal material in Menshi huwei formula， extraction rates of both are increased slightly.

OBJECTIVE： To establish a method for simultaneous determination of α-pinene， β-pinene， limonene and α-terpineol in volatile oil of Forsythia suspensa. METHODS： GC method was adopted. The determination was performed on HP-5 capillary column through temperature-programmed route. The inlet temperature was 230 ℃， and detector temperature was 250 ℃； split sampling was applied （split ratio of 8 ∶ 1）； the air flow rate was 300 mL/min， the hydrogen flow rate was 30   mL/min， the tail gas flow rate was 30 mL/min， and the injection volume was 1 μL. Using limonene as internal reference， relative correction factors of α-pinene， β-pinene and α-terpineol were established， and the reproducibility of relative correction factors were investigated by using different chromatographs and columns， and chromatographic peak location of components was measured. The contents of above components were calculated with QAMS， and then compared with the results of external standard method. RESULTS： The linear range of α-pinene， β-pinene， limonene and α-terpineol were 16.5-990.0， 38.1-2 287.5， 8.2-491.2， 2.4-142.5  μg/mL， respectively （r≥0.999 1）. RSDs of precision， reproducibility and stability tests were all lower than 3％ （n＝6）. Average recoveries were 99.7％-105.5％（RSD＜4％，n＝9）. Compared with limonene （1.00），the average relative correction factors of α-pinene， β-pinene and α-terpineol were 0.91，0.86 and 1.11（n＝3）； relative retention time were 0.69-0.74， 0.81-0.86， 1.25-1.35（RSD＜3％，n＝3）. By using different chromatographs and columns， RSDs of relative correction factors were 0.21％-4.65％（n＝6）. Compared with external standard method， determination results of above 4 components were consistent （the absolute value of relative error were all less than 7％）. CONCLUSIONS： QAMS can be used for simultaneous determination of 4 kinds of effective components in volatile oil from F. suspensa.

OBJECTIVETo observe the herbal effects on hyperthyroidism patients with syndrome of hyperactivity of liver-Yang by method for calming the liver and suppressing Yang and investigate its effects on the lymphocyte protein expression. This approach may lay a foundation for the further investigation of the curative mechanisms of calming the liver and suppressing Yang treatment.

METHODA total of 48 hyperthyroidism patients with syndrome of hyperactivity of liver-Yang were randomly divided into treatment group and control group. The treatment group was treated by method for calming the liver and suppressing Yang in accordance with traditional Chinese medicine (TCM) and the control group with thiamazole tablets for three periods of treatment The therapeutic effects, the score of TCM symptom, electrocardiogram (P wave), thyroid hormones and ultrasound were observed in both groups before and after the treatment. The side effects in the treatment course were observed in both groups. The level of differential protein expression was analyzed by two-dimensional electrphoresis and matrix assisted laser desorption/ionizaton time-of-flight mass spectrometry.

RESULTThe treatment group has the effect on stepping down the heart rate, cutting down the P wave amplitude changes, regulating the level of thyroid hormones and decreasing the volume of thyromegaly. There are not statistically significant between the treatment group and control group. However, the treatment group has obviously better effect on regulating TCM symptom and decreasing the side reaction than the control group (P<0.05). There are not statistically significant on the total effective between the treatment group and control group. The average spots in lymphocyte for normal people, before and after treating hyperthyroidism patients with syndrome of hyperactivity of liver-Yang were (429 +/- 31), (452 +/- 28) and (437 +/- 36) spots respectively. Eight down-regulated protein expressions and 11 up-regulated protein expressions were obtained in the hyperthyroidism patients with syndrome of hyperactivity of liver-Yang and normal people. Five strengthened expressions of protein were also obtained in 8 down-regulated expressions of protein and 8 lower expressions of protein in 11 up-regulated expressions of protein before and after treating the migraine patients with syndrome of hyperactivity of liver-Yang. Ten of the total 8 differential protein spots were successfully identified by MALDI-TOF-MS. The functions of these proteins were involved in metabolism associated, transportation, antioxidation, sigal transduction and immume associated protein, etc. according to information provided by NCBI and MSDB database.

CONCLUSIONIn this study, the TCM complex prescription with herbs for calming the liver and suppressing Yang can regulate the thyroid hormones, improves TCM symptoms, and decrease the adverse reaction. It can possibly regulate lymphocyte protein expression.

Adult ; Aged ; Databases, Protein ; Drugs, Chinese Herbal ; adverse effects ; pharmacology ; therapeutic use ; Electrocardiography ; Female ; Gene Expression Regulation ; drug effects ; Humans ; Hyperthyroidism ; drug therapy ; metabolism ; pathology ; physiopathology ; Liver ; drug effects ; pathology ; physiopathology ; Lymphocytes ; drug effects ; metabolism ; Male ; Middle Aged ; Proteins ; metabolism ; Thyroid Hormones ; metabolism ; Treatment Outcome ; Yin-Yang ; Young Adult

Objective To explore the feasibility and efficiency of the treatment of lumbar degenerative diseases after transforaminal lumbar interbody fusion(TLIF)and posterolateral fusion(PLF)procedures in which unilateral pedicle screw fixation was used.Methods From December 2006 to August 2008,78 cases with the lumbar degenerative diseases who received lumbar posterolateral fusion were analyzed.There were 48 cases of which underwent TLIF and PLF procedures with unilateral pedicle screw fixation(unilateral group),including 25 males and 23 females with an average of 47.6 years;and 30 cases of which underwent TLIF and PLF procedures with bilateral pedicle screw fixation(bilateral group),including 21 males and 9 females with an average of 50.5 years.The clinical effects between the two groups were evaluated with Oswestry disability index and visual analogue score(VAS)index.The operation time,blood loss,fusion rates and intervertebral collapse rates were also compared.Results Oswestry disability index,low back pain VAS index and skelalgia VAS index in both groups showed statistical significance between preoperation and 3 months,or 3 months and 1 year postoperatively.There was no difference in score improvement between the two groups.There were difference in operation time,blood loss and cost of hospitalization between unilateral and bilateral group.The former was lower.There was no difference in postoperative length of stay between the two groups.The fusion rate of unilateral group and bilateral group were 91.7%(44/48)and 93.3%(28/30),respectively.Conclusion Auto graft combined with unilateral pedicle screw fixation provids better spinal instant stability.TLIF and PLF with unilateral pedicle screw fixation was a satisfactory method in treating degenerative disease of lumbar vertebrae.

The complete genome sequence of transmissible Gastroenteritis virus (TGEV) strain TS, previously isolated from Gansu province, was cloned and compared with published sequence data from other TGEV strains.Phylogenetic tree analysis based on the amino acid and nucleotide sequences of the S gene showed that the TGEV strains were divided into 3 clusters. TGEV TS showed a close evolutionary relationship to the American Miller cluster but had a 5' non-translated region (NTR) sequence closely related to the American Purdue cluster.Continued culture in different cell types indicated that TGEV TS virulence could be attenuated alter fifty passages in Porcine kidney (PK-15) cells, and that the Porcine kidney cell line IB-RS-2 (IBRS) was not suitable for culture of the TGEV strain TS.

Objective To explore the deficit features of verbal working memory of dyslexia children by comparing with normal children. Methods Seventeen normal children and fifteen dyslexia children finished the classic verbal n-back task which involved three memory updating difficulty levels. Results Dyslexia children ((81.7±7.0)%,(70.8±12.0)%,(60.0±7.0)%)performed worse than NC ((88. 6 ±6. 0)% , (83. 5 ± 7.0)% ,(75.6 ±9.0)%)in all memory updating levels(n = 0,1 ,2) (P<0.01). In the performances related to memory updating operating,when the difficulty of the updating rised dyslexia children(25. 9 ± 12. 0)% revealed greater decline than normal children(14.2 ± 12.0)% (P<0.01). Conclusion Dyslexia children demonstrate the deficit in the basic level of working memory, which was showed as the bad performance of n-back task in updating level zero, and also they had problems in memory updating,which was showed as the greater updating effect. Memory updating was regarded as the advanced procession of verbal working memory.