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.
Animals ; Clonorchis sinensis/genetics* ; Metacercariae/genetics* ; Recombinases ; Fresh Water ; Fishes ; DNA ; Fish Diseases/diagnosis*

Objective:To evaluate the efficacy of subomohyoid anterior suprascapular nerve block (SSNB) in the patients undergoing arthroscopic shoulder surgery with general anesthesia.Methods:Sixty patients of either sex, aged 18-64 yr, with body mass index of 18-30 kg/m 2, of American Society of Anesthesiologists physical status Ⅰor Ⅱ, scheduled for elective arthroscopic shoulder surgery, were divided into 2 groups ( n=30 each) using a random number table method: SSNB group (S group) and interscalene brachial plexus block group (I group). Before induction, 0.375% ropivacaine hydrochloride 15 ml was injected between C 5-C 6 nerve roots in group I and around the anterior suprascapular nerve in group S under ultrasound guidance.Diaphragmatic excursion, occurrence and degree of diaphragmatic paralysis, decrease in SpO 2, dyspnea, Horner syndrome and sensory block in the C 5-T 1 dermatomes were assessed at 30 min after injection.The intraoperative consumption of remifentanil, extubation time, and length of post-anesthesia care unit stay were recorded.Quality of Recovery-15 score for patient′s satisfaction with analgesia, effective pressing frequency of analgesic pump, requirement for rescue analgesia, nausea and vomiting and nerve block-related complications within 24 h after surgery were recorded. Results:Compared with group I, the incidence of diaphragmatic paralysis was significantly decreased, the degree of diaphragmatic paralysis was reduced, diaphragmatic excursion was increased, the amplitude of decrease in SpO 2 was reduced, the incidence of dyspnea and Horner syndrome was decreased, extubation time was shortened ( P<0.05), and no significant change was found in the incidence of sensory block in the C 5-T 1 dermatomes, intraoperative consumption of remifentanil, effective pressing frequency of analgesic pump, requirement for rescue analgesia, score for patient′s satisfaction with analgesia, incidence of nausea and vomiting, length of post-anesthesia care unit stay, or Quality of Recovery-15 score in group S ( P>0.05). Conclusions:The subomohyoid anterior SSNB not only provides reliable perioperative analgesia, but also reduces the risk of diaphragmatic paralysis when used in the patients undergoing arthroscopic shoulder surgery with general anesthesia.

Objective:To explore the influencing factors of bowel preparation quality in hospitalized elderly patients, and to find the appropriate waiting time from the end of bowel preparation to the beginning of colonoscopy.Methods:Baseline and clinical data of elderly patients over 60 years old who underwent colonoscopy in the Tenth People′s Hospital, Tongji University from February 2021 to August 2021 were collected. Multivariate analysis was used to screen the factors that might affect the quality of bowel preparation in hospitalized elderly patients. Patients were grouped according to waiting time before colonoscopy. After eliminating confounding factors using propensity matching analysis, the difference of bowel preparation quality among groups was compared.Results:251 patients were included in the study. Multivariate analysis revealed that, hypertension ( OR=3.530, 95% CI 1.295-9.618), chronic constipation ( OR=3.302,95% CI 1.132-9.632), dietary compliance ( OR=0.161, 95% CI 0.070-0.371), medication and drinking water compliance ( OR=0.167, 95% CI 0.070-0.397), exercise compliance after medication ( OR=2.245, 95% CI 1.040-4.845), The frequency of defecation after medication ( OR=0.446, 95% CI 0.308-0.647) and waiting time ( OR=0.537, 95% CI 0.387-0.745) were important factors affecting the quality of bowel preparation in hospitalized elderly patients ( P<0.05). There were differences in bowel preparation quality between groups of waiting times. The overall quality of bowel preparation in 120-180 min group was significantly better than that in 241-300 min group, 301-360 min group and>360 min group ( P<0.05). The overall quality of bowel preparation in 181-240 min group was better than that in >360 min group ( P<0.05). There were no significant differences among other groups( P>0.05). The scores of cecum and ascending colon were the best in 120-180 min group, and the cleanliness of descending colon, sigmoid colon and rectum was significantly higher in 241-300 min group, 301-360 min group and > 360 min group. The scores of descending colon, sigmoid colon and rectum showed that the intestinal preparation quality of 181-240 min group was better than that of 301-360 min group and > 360 min group. Conclusions:The best examination time for elderly patients is about 180 minutes after bowelpreparation. Medical workers should flexibly guide the medication time to ensure that patients are in the best clean state of intestinal tract during examination.

OBJECTIVE：To establish a meth od for the simultaneous determination of 7 active components in Mori Australis Cortex and Mori Cortex from different sources in Chongqing area ，so as to provide reference for improving the quality control standards of Mori Australis Cortex and Mori Cortex and comparing the equivalence of their quality. METHODS ：HPLC method was used to determine the contents of neochlorogenic acid ，mulberroside A ，chlorogenic acid ，astragalin，kaempferol，morusin and isoquercetin in 58 batches of Mori Australis Cortex and Mori Cortex. The chromatographic column was Diamonsil C 18 with mobile phase consisted of 0.1％ formic acid solution-acetonitrile （gradient elution ） at the flow rate of 1.0 mL/min. The detection wavelength was 280 nm，column temperature was 30 ℃，and the injection volume was 10 μL. Using SPSS 22.0 software， independent sample t-test，principal component analysis and cluster analysis were used to analyze the content difference of the above-mentioned 7 active components in Mori Australis Cortex and Mori Cortex. RESULTS ：There was a good linear relationship between the peak area and the concentration of the above 7 active components （r≥0.999 0）. The RSDs of precision ，stability（24 h），repeatability，durability and recovery were less than 3％. The average contents of neochlorogenic acid ，mulberroside A ， chlorogenic acid ， astragalin， kaempferol， morusin and 023-58576130。E-mail：1025473978@qq.com isoquercetin in Mori Australis Cortex were 0.304，22.462， 1.730，1.308，1.593，2.842 and 0.657 mg/g，respectively. Those of Mori Cortex were 0.305，22.995，2.486，2.438， 2.916，4.158 and 1.264 mg/g，respectively. The results of independent sample t-test showed that only the content of kaempferol in the above 7 active components of Mori Australis Cortex and Mori Cortex had significant difference （P＜0.05）. The results of principal component analysis and cluster analysis showed that there was no significant difference in the contents of above 7 active components between Mori Australis Cortex and Mori Cortex. CONCLUSIONS：The established HPLC method is simple ，sensitive and accurate ，which can provide a reference for improving the quality control standard of Mori Australis Cortex and Mori Cortex. Mori Australis Cortex and Mori Cortex have certain quality equivalence in main active components ，and the Mori Australis Cortex from M. australis and M. cathayana can be used as a substitute for the Mori Cortex.

Objective:To analyze the clinical characteristics and prognosis of pregnant women with hemorrhagic fever with renal syndrome (HFRS).Methods:A total of 11 pregnant women with HFRS admitted to The Second Affiliated Hospital of Xi′an Jiaotong University (four cases), The Second Affiliated Hospital of Air Force Medical University (four cases), The First Affiliated Hospital of Xi′an Jiaotong University (one case) and Central Hospital of Xianyang City (two cases) between November 2009 and February 2019 were included as the study group, and 24 age-matched non-pregnant women with HFRS were selected as the control group. The age, complications, clinical classification and laboratory indexes of the two groups were analyzed retrospectively, and the clinical outcomes of pregnant women and their fetuses in the study group were followed up. The data between two groups were compared using Mann-Whitney U test or chi-square test. Results:Patients in the study and control groups were 29 (22, 33) and 32 (24, 37) years old, respectively. Seven of 11 patients in study group were severe and critical cases, which was significantly higher than that in the control group (16.7%(4/24), χ2=7.722, P=0.015). In the study group, 10 patients had hypervolemic syndrome, 10 patients had pulmonary edema and six patients had overlapping hypotension shock phase and oliguria phase, which were all higher than those in the control group ((2/24, 8.3%), (2/24, 8.3%) and (2/24, 8.3%), respectively; χ2=22.828, 22.828 and 9.135, respectively, all P<0.01). Compared with the control group, the pregnant patients in study group had a higher urea nitrogen maximum and serum creatinine maximum, and the differences were both statistically significant ( Z=-2.453 and -2.336, respectively, both P<0.05), while they had a lower serum albumin minimum, hemoglobin maximum and hemoglobin minimum, and the differences were all statistically significant ( Z=-3.742, -3.350 and -4.034, respectively, all P<0.01). All pregnant women with HFRS recovered. Nine pregnant women gave birth to nine healthy infants. All of them received breastfeeding and the feeding duration were more than six months. No abnormal growth and development were found during an average follow-up of three years. Conclusions:Pregnancy can aggravate the severity of HFRS, and pregnant women have higher risk of the multiple stages overlap and the complications such as hypervolemic syndrome and acute pulmonary edema. After recovery from HFRS, mother may carry to full-term pregnancy.

Objective:To investigate the inhibitory effect and molecular mechanism of microRNA-30d (miR-30d) in the process of proliferation, migration and invasion of malignant mesothelioma cell line MSTO-211H.Methods:In April 2017, the human MSTO-211H cells was used to establish miR-30d overexpressed MSTO-211H cell model by transfection of miR-30d mimics. The qRT-PCR was performed to detect the expression level of miR-30d in the cells transfected miR-30d mimics. The effects of miR-30d on the proliferation, apoptosis, migration and invasion of MSTO-211H cells were analyzed by CCK-8 experiment, flow cytometry, cell scratch experiment and Transwell method.Results:After transfection of miR-30d, the expression level of miR-30d in the MSTO-211H+miR-30d cells group was significantly higher than MSTO-211H+miR NC cells group ( P<0.01) . The cell activity of MSTO-211H+miR-30d group (105.13%±2.35%) was significantly lower than MSTO-211H+miR NC cells group (115.40%±1.35%) , and the level of apoptosis (3.97%±0.36%) was significantly higher than MSTO-211H+miR NC cells group (1.47%±0.10%) ( P<0.01) . The relative migration areas at 12 and 24 h of MSTO-211H+miR-30d cells group (9.35±3.16 μm 2 and 58.19±1.82 μm 2) were significantly lower than MSTO-211H+miR NC cells group (54.42 ±5.26 μm 2 and 88.32 ±1.96 μm 2) ( P<0.01) . Compared with the MSTO-211H+miR NC cells group, the numbers of cell migration and cell invasion were reduced in the MSTO-211H+miR-30d cells group ( P<0.01) . Conclusion:miR-30d can regulate the progression of malignant pleural mesothelioma by inhibiting the proliferation, apoptosis, migration and invasion of MSTO-211H cells.

Objective:To investigate the inhibitory effect and molecular mechanism of microRNA-30d (miR-30d) in the process of proliferation, migration and invasion of malignant mesothelioma cell line MSTO-211H.Methods:In April 2017, the human MSTO-211H cells was used to establish miR-30d overexpressed MSTO-211H cell model by transfection of miR-30d mimics. The qRT-PCR was performed to detect the expression level of miR-30d in the cells transfected miR-30d mimics. The effects of miR-30d on the proliferation, apoptosis, migration and invasion of MSTO-211H cells were analyzed by CCK-8 experiment, flow cytometry, cell scratch experiment and Transwell method.Results:After transfection of miR-30d, the expression level of miR-30d in the MSTO-211H+miR-30d cells group was significantly higher than MSTO-211H+miR NC cells group ( P<0.01) . The cell activity of MSTO-211H+miR-30d group (105.13%±2.35%) was significantly lower than MSTO-211H+miR NC cells group (115.40%±1.35%) , and the level of apoptosis (3.97%±0.36%) was significantly higher than MSTO-211H+miR NC cells group (1.47%±0.10%) ( P<0.01) . The relative migration areas at 12 and 24 h of MSTO-211H+miR-30d cells group (9.35±3.16 μm 2 and 58.19±1.82 μm 2) were significantly lower than MSTO-211H+miR NC cells group (54.42 ±5.26 μm 2 and 88.32 ±1.96 μm 2) ( P<0.01) . Compared with the MSTO-211H+miR NC cells group, the numbers of cell migration and cell invasion were reduced in the MSTO-211H+miR-30d cells group ( P<0.01) . Conclusion:miR-30d can regulate the progression of malignant pleural mesothelioma by inhibiting the proliferation, apoptosis, migration and invasion of MSTO-211H cells.

OBJECTIVE：To estab lish a me thod for simultaneous determination of morroniside ，loganin，echinacoside and acteoside in Huanshao capsules. METHODS ：HPLC method was adopted. The determination was performed on Zhongpuhong RD-C18 column with mobile phase consisted of acetonitrile- 0.1％ formic acid solution （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was set at 240 nm （morroniside，loganin） and 330 nm （echinacoside，acteoside）. The column temperature was set at 35 ℃，and sample size was 10 μL. RESULTS：The linear range were 5.29-105.80 μg/mL for morroniside， 4.49-89.88 mg/L for loganin ，16.26-325.25 mg/L for echinacoside and 16.31-326.25 mg/L for acteoside ，r values were 0.999 9. RSDs of precision ，stability （24 h），reproducibility and durability tests were all lower than 2.0％ . The recoveries were 94.34％ -96.23％（RSD＝0.81％ ，n＝6），97.04％ -98.89％（RSD＝0.73％ ，n＝6），96.23％ -98.08％（RSD＝0.82％ ，n＝6）， 95.40％-98.47％（RSD＝1.23％，n＝6），respectively. The contents of above 4 components in 11 batches of Huanshao Capsules were 0.190-0.704，0.439-0.857，2.723-4.475 and 0.589-1.035 mg/g，respectively. CONCLUSIONS ：Established method is specific ， precise and can be used for content determination of 4 components in Huanshao capsules.

Objective:This study aims to investigate the expression of E3 ubiquitin-ligase (WWP1) in chronic lymphocytic leukemia (CLL) patients and analyze its correlation with clinical prognostic indicators (TP53, CD38, IGHV mutation) and its prognostic value.Methods:A total of 48 CLL patients and 9 age-matched normal subjects were enrolled in the study. The WWP1 expression was detected by SYBR Green-based real-time PCR, and the clinical relationship was analyzed by GraphPad Prism software.Results:The WWP1 median expression was 0.007 (95% CI 0.005-0.010) in the normal control group and 0.031 (95% CI 0.019-0.044) in the CLL group ( P<0.001) . A sub-groups analysis implicated a statistically significant result ( P=0.022) , showing that the median time from a relatively high and low transcription level of WWP1 to the first treatment was 24 months and 35 months, respectively. Positive CD38 and ZAP-70 expressions were associated with a higher WWP1 expression ( P=0.012 and 0.029, respectively) . Conclusion:An abnormal WWP1 mRNA expression was found in CLL patients with significant correlation with ZAP-70 and CD38 expressions, and WWP1 may become a new supplement of CLL prognostic markers.

Objectives To evaluate the effectiveness and safety of peramivir trihydrate in patients with influenza.Methods This was a randomized,double-blind,double-dummy,placebo and positive control,multicenter clinical trial,comparing peramivir trihydrate with oseltamivir and placebo.The inclusive criteria were 15-70 years old,onset within 48 h,positive rapid influenza antigen test,and febrile(＞38℃) accompanied with at least two associated symptoms.The severe cases complicated with chronic pulmonary and cardiac diseases,malignancies,organ transplantation,hemodialysis,uncontrolled diabetes,immunocompromised status,pregnancy and coexistence of bacterium infections were excluded.All patients were randomized 2:2:1 to receive peramivir,oseltamivir and placebo respectively.The primary endpoint was the disease duration,the secondary endpoints included time to normal axillary temperature and normal living activities,viral response,and adverse effects.Results Following informed consent,133 patients were included in this study.Four patients were exclude due to missing medical records,not fitting inclusion or exclusion criteria and poor compliance.A total of 129 patients were finally analyzed,including 49 cases,54 cases and 26 cases in peramivir group,oseltamivir group and placebo group.The median disease duration were 96 (76,120)hours,105(90,124) hours,and 124 (104,172)hours in three groups respectively(P＞0.05).The time to normal axillary temperature,normal living activities and viral response were not significantly different in three groups(P＞0.05).Conclusion The value of antiviral therapy in patients with mild influenza needs to be further determined.