ObjectiveRapid and accurate identification of body fluid traces at crime scenes is crucial for case investigation. Leveraging the speed and sensitivity of nucleic acid detection technology based on SHERLOCK, our research focuses on developing a peripheral blood SHERLOCK-HBA detection system to detect mRNA in forensic practice. MethodsShort crRNA fragments targeting the blood-specific mRNA gene HBA were designed and screened, alongside RPA primers. Optimal RPA primers were selected based on specificity and amplification efficiency, leading to the establishment of the RPA system. The most efficient crRNA was chosen based on relative fluorescence units (RFU) generated by the Cas protein reaction, and the Cas protein reaction system was constructed to establish the SHERLOCK-HBA detection method. The RPA and Cas protein reaction systems in the SHERLOCK detection system were then individually optimized. A total of 79 samples of five body fluids were tested to evaluate the method’s ability to identify blood, with further verification through species-specific tests, sensitivity tests, mixed spots detection, aged samples, UV-irradiated samples, and actual casework samples. ResultsThe SHERLOCK reaction system for the peripheral blood-specific marker HBA was successfully established and optimized, enabling detection within 30 min. The method demonstrated a detection limit of 0.001 ng total RNA, better than FOB strip method and comparable to RT-PCR capillary electrophoresis. The system could detect target body fluids in mixed samples and identify blood in samples stored at room temperature for three years and exposed to UV radiation for 32 h. Detection of 11 casework samples showed performance comparable to RT-PCR capillary electrophoresis. ConclusionThis study presents a CRISPR/Cas-based SHERLOCK-HBA detection system capable of accurately, sensitively, and rapidly identifying blood samples. Introducing CRISPR/Cas technology to forensic body fluid identification represents a significant advancement in applying cutting-edge molecular biology techniques to forensic science.The method’s simplicity, shorter detection time, and independence from specialized equipment make it promising for rapid blood sample identification in forensic cases.

Objective To explore the effectiveness of different intervention strategies in response to outbreaks of emerging highly pathogenic respiratory infectious diseases(RIDs)in mega-city in China,and provide decision-ma-king basis for effective response to emerging RIDs.Methods A susceptible-exposed-infectious-recovered(SEIR)transmission dynamics model was constructed,referencing to and combining the pathogenicity and infectivity para-meters of previous emerging RIDs.The outbreak of emerging highly pathogenic RIDs with low,moderate,and high infectivity in a mega-city with a population of 10 million in China was simulated,the development of the epidemic within 100 days after implementing different combinations of non-pharmaceutical interventions(NPIs)in response to the outbreak was compared.Results When highly pathogenic RIDs outbreak occurred,and if its infectivity was low(R0 was about 1.5),it was unnecessary to adopt strict NPIs to control epidemic.If its infectivity was moderate(R0 was about 6),different intensities of NPIs were needed based on its existing infection scale.When the initial num-ber of infected cases was 50,moderate-intensity NPIs could keep the infection and death at a low level within 100 days,and the required bed number in hospital for cases could be kept below the national average reserve level.But when the scale of infection exceeded 100 cases,high-intensity NPIs were needed to control the development of the epidemic.In the case of extremely strong infectivity(R0 was about 10),regardless of the scale of infection,only immediate high-intensity NPIs could control the epidemic,infection and death scale.Conclusion In case of out-breaks of highly pathogenic RIDs,adopting appropriate NPIs as early as possible based on their epidemiological characteristics and infection scale is necessary to minimize the harm to the population.

Objective:To analyze measurement results of serum allergen-specific immunoglobulin E (IgE) in patients with eczema/dermatitis.Methods:A retrospective analysis was conducted on serum allergen-specific immunoglobulin E (IgE) levels in 3 051 patients with eczema/dermatitis, who visited the allergy clinic of Huashan Hospital from April 1, 2021 to March 31, 2022. The serum allergen-specific IgE level was detected by using the Phadia allergen detection system, and positive rates of allergens were calculated to determine common inhaled allergens and food allergens in patients with eczema/dermatitis. Comparisons of enumeration data between groups were performed by chi-square test.Results:Among the 3 051 patients with eczema/dermatitis, there were 1 412 with atopic dermatitis and 1 639 were other eczema/dermatitis. Detection of serum allergen-specific IgE showed that 1 629 (53%) patients were positive for allergens, and the number of positive allergen-specific IgEs in each patient was 3.0 ± 1.6. The top 3 common inhaled allergens in patients with eczema/dermatitis were Dermatophagoides farinae (904/1 522, 59%) , Dermatophagoides pteronyssinus (891/1 513, 59%) and Alternaria alternata (206/1 068, 19%) , and the top 3 common food allergens were shrimps (251/1 432, 18%) , egg white (165/992, 17%) and cow milk (149/994, 15%) . Among the 3 051 patients, 25 (1%) were aged < 2 years, 571 (19%) aged 2 - 12 years, 285 (9%) aged 12 - 18 years, and 2 170 (71%) were aged > 18 years. The most common food allergens were both egg white in the age groups of < 2 years and 2 -12 years (77%, 37%, respectively) , and were both shrimps in the age groups of 12 - 18 years and > 18 years (31%, 17%, respectively) . Dermatophagoides pteronyssinus and Dermatophagoides farina were the top 2 common inhaled allergens in all age groups, with the positive rate ranging from 36% to 84%; in addition, the positive rate of molds was relatively high in the age group of 2 - 12 years (mold mixture: 37%; Alternaria alternata: 27%) . From April 2021 to March 2022, the positive rate of outdoor allergens ranged from 10% to 15% among outpatients in every month; the positive rates of tree pollen and grass pollen increased from April 2021, and peaked in October 2021. The patients with atopic dermatitis showed a significantly increased positive rate of allergens (73%) compared with those with other eczema/dermatitis (37%, χ2 = 389.36, P<0.001) , and the rank of common allergens in the patients with atopic dermatitis was basically the same as that in those with eczema/dermatitis. Conclusions:The common allergens were Dermatophagoides farina, Dermatophagoides pteronyssinus and Alternaria alternata in the patients with eczema/dermatitis. Food allergy was more common in infant patients, and inhalation allergy was more common in child, adolescent and adult patients. The positive rate of allergen-specific IgEs was markedly higher in the patients with atopic dermatitis than in those with other eczema/dermatitis.

Aim To investigate the effect of microinjection of EX527, a selective SIRT1 antagonist, into the ventrolateral orbital cortex (VLO) on morphine-induced conditioned place preference (CPP), and to explore the role of CREB/BDNF in it. Methods The cannulas were implanted bilaterally in the VLO of rats by brain stereotaxis surgery, and the model of morphine-induced CPP was established. The behavioral experiment consisted of four stages:habituation (d 1), pre-test (d 2-4), conditioning training (d 5-14) and test (d 15). At the stage of conditioning training, EX527 (1 μL, 5 g·L

Objective:To investigate the clinical features and risk factors of infection in patients with heat stroke (HS).Methods:The patients with HS who were admitted to the emergency departments of seven hospitals, including West China Hospital of Sichuan University, the Second Affiliated Hospital of Chengdu Medical College and so on, between July 01, 2020 and September 30, 2022 were retrospectively analyzed. The basic characteristics and laboratory test results of the patients were collected. The patients were divided into infected group and non-infected group according to the etiological results, and the differences in clinical characteristics between the different groups were compared. Multivariate logistic regression was used to screen variables, and the predictive model was established and ROC curve was performed to compare the predictive efficiency with SOFA score.Results:During the study period, a total of 183 patients with HS were included, and 156 patients were finally included for analysis according to the inclusion and exclusion criteria. Among them, 58 cases (37.2%) were in the infected group. The mortality of infected group was significantly higher than that of non-infected group (41.4% vs. 25.5%, P < 0.05). Compared with the patients in non-infected group, the infected gourp serum procalcitonin, direct bilirubin, alanine transaminase (ALT), creatinine, lactate dehydrogenase (LDH), creatine kinase isoenzymes, myoglobin, activated partial thromboplastin time (APTT) and respiratory support ratio at admission were significantly higher and albumin level was lower in infected group (all P < 0.05). Multivariate logistic regression analysis showed that myoglobin, ALT, APTT and respiratory support required at admission were independent risk factors for HS co-infection. A prediction model was constructed based on these four indicators, and the area under ROC curve of the prediction model was 0.846, which was better than SOFA score (0.732, P < 0.05). Conclusions:The prognosis of patients with heat stroke complicated with infection were obviously poor. Myoglobin, ALT, APTT and need for respiratory support at admission are risk factors for HS complicated with infection. The prediction model based on these four indicators is better than SOFA score for the early identification of the HS patients with infection.

Objective: To analyze the epidemiological characteristics and influencing factors of COVID-19 confirmed cases with viral nucleic acid re-positive in anal and/or throat swabs after discharge during the domestic imported epidemic stage in Guangdong Province in early 2020. Methods: The COVID-19 confirmed cases with the onset time before March 1, 2020 in Guangdong Province were collected to analyze the demographic data, epidemiological characteristics, and specimen collection and testing data after discharge. Logistic regression model was used for influencing factors analysis of re-positive cases. Results: A total of 1 286 COVID-19 confirmed cases were included, the M(Q₁,Q₃) of age was 44(32,58)years, 617 cases were male, 224 cases were re-positive in anal and/or throat swabs with the re-positive rate 17.42%. The M(Q₁,Q₃) of age of re-positive cases was 35(23, 50) years, which was younger than that of re-negative cases age was those 46(33, 59) years (P<0.001). With the increase of age, re-positive rate decreased (χ²_trend=52.73, P<0.001). 85.27% (191/224) of re-positive cases were found in 14 d after discharge, the duration time of re-positive status was 13(7, 24) d, and 81.69% (183/224) of re-positive cases were re-tested negative in 28 d after re-positive date. No fever and other symptoms had been observed among re-positive cases during the whole follow-up. No secondary infectious cases had been found among close contacts after 14 d of centralized isolation and sampling screening. Univariate logistic regression model analysis revealed that the influencing factors of the re-positive cases included age, occupation, clusters, clinical types, and admission time. Multivariate logistic regression model analysis revealed that age was an independent risk factor. Conclusions: SARS-CoV-2 viral nucleic acid re-positive is found in COVID-19 confirmed cases after discharge in Guangdong Province. Most re-positive cases are confirmed among 14 d after discharge and re-test to negative among 28 d after re-positive date. Age is an risk factor for re-positive cases after discharge.
COVID-19 ; China/epidemiology* ; Epidemics ; Humans ; Male ; Nucleic Acids ; SARS-CoV-2

Objective:To analyze and discuss the application value and teaching effect of simulate patients (SP)-assisted problem-based learning (PBL) combined with team-based learning (TBL) mode in pediatric first aid teaching, in order to provide a new reference for the reform of standardized residency training and teaching mode of pediatrics in the new era in China.Methods:A total of 103 residents who participated in the standardized training in the Department of Pediatrics of Hunan Children's Hospital from January 2018 to December 2020 were selected as the research objects, and they were divided into the control group (50 cases) and the experimental group (53 cases). The control group adopted the previous teaching mode of residents, while the experimental group adopted the SP-assisted PBL combined with TBL mode. In this study, SPSS 22.0 software was used for t test and chi-square test. Results:The examination scores of the experimental group in the theoretical knowledge of pediatric first aid, clinical thinking and skills, and application of pediatric first aid skills were higher than those in the control group [(86.98±3.42), (85.69±5.13), (89.62±4.75)] vs. [(77.23±4.16), (81.16±3.96), (76.54±5.78)], with statistically significant differences between the two groups ( P<0.05). The questionnaire survey showed that the clinical thinking and pediatric emergency skills, doctor-patient communication ability, teamwork ability and other aspects of the experimental group were significantly better than those of the control group ( P<0.05). Conclusion:The SP-assisted PBL combined with TBL mode can produce positive effects in the standardized residency training of pediatrics, which will help residents better master pediatric first aid skills, improve their clinical comprehensive capabilities, and be more conducive to improving the quality of pediatric residency training, which is worth promoting in the new era.

This study aims to establish the high-performance liquid chromatography(HPLC) fingerprints of different batches of Notoginseng Radix et Rhizoma, determine their pharmacodynamic indexes of promoting blood circulation, and explore the spectrum-effect relationship between the chemical components of Notoginseng Radix et Rhizoma and the efficacy of promoting blood circulation. Firstly, the HPLC fingerprints of different batches of Notoginseng Radix et Rhizoma were established. Then, the pharmacodynamic indexes were determined after the capillary coagulation experiment and the cerebral ischemia-reperfusion in rats, including capillary coagulation time, percentage of cerebral ischemic area, cerebral water loss rate, and brain-body index. Afterward, the partial least-squares method was used to explore the spectrum-effect relationship between the chemical components of Notoginseng Radix et Rhizoma and the pharmacodynamic indexes. The results showed that this study successfully established the HPLC fingerprints of different batches of Notoginseng Radix et Rhizoma, found 23 common peaks, and identified 12 of them, all of which were saponins. The method was proved stable and reliable. Both the capillary coagulation experiment and the middle cerebral artery occlusion(MCAO)-induced cerebral ischemia-reperfusion experiment on rats revealed that there were obvious differences in the pharmacodynamic indexes of different batches of Notoginseng Radix et Rhizoma. The relationships between 23 common components of Notoginseng Radix et Rhizoma in different batches and the pharmacodynamic indexes were discussed by means of spectrum-effect correlation analysis, of which 17 components had positive effects while 6 components had negative effects on the pharmacodynamic indexes. This study provides a certain reference basis for the clinical rational use and quality control of Notoginseng Radix et Rhizoma.
Animals ; Blood Coagulation ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Quality Control ; Rats ; Rhizome ; Saponins

Objective::To investigate the distribution status of medicinal plants in the wild areas of Russian Caucasus and Altai, and clarify the types and efficacy information of medicinal plants in the area, so as to dig deep into new resources and new functions of medicinal plants in the countries along the Belt and Road. Method::Medicinal plants in the wild were searched and collected to make waxy specimens, and sent back to the country to extract the total DNA of the leaves of the leaves. Internal Transcribed Spacer(ITS)sequence universal primers were used for Polymerase Chain Reaction (PCR)amplification. The PCR products were sent for the two-way sequencing, and the sequencing results are spliced by software according to National Center for Biotechnology Information(NCBI). The same ITS sequence of the highest similarity species obtained by database BLAST was analyzed by DNAman software to identify the ITS sequence of the species and the ITS sequence of the same species. The MEGA 7 software was used as the phylogenetic tree, and the Kimura-2 parameter genetic distance was used to construct the neighbor joining(NJ) phylogenetic tree by the neighbor-joining method. The confidence of each branch of the development tree was tested by the bootstrap test method. A total of 2 000 cycles were performed, and the results were identified based on the clustering results. On this basis, the key medicinal plants in the Russian Caucasus and Altay wild areas were summarized and analyzed. Result::After BLAST alignment in NCBI database, the ITS sequences of each specimen were clustered with the login sequences on the NCBI database, which were separated from the outer group. The species classification of the specimens to be identified was determined by combining the characteristics of the specimens. A total of 51 plants were identified from the specimens collected in the field, covering 44 genera of 17 families, and 29 plants had clear efficacy records. The National Drug List of the Russian Federation and the Chinese Pharmacopoeia were retrieved to summarize commonly used medicinal plants in China and conclude that 20 kinds of Chinese and Russian common medicinal materials have different medicinal effects in local areas. This study has a reference significance for expanding the scope and clinical experience of traditional Chinese medicines, and provides a basis for strengthened local species conservation, development and utilization of wild medicinal plant resources.