Objective To study a nucleic acid extraction method suitable for detecting methicillin‐resistant Staphylococcus aureus (MRSA) by PCR method .Methods Under different incubation conditions ,MRSA was cracked by lysozyme ,lysostaphin or chel‐ex100R resin for obtaining DNA ,then the target gene was detected by using the PCR method .Results DNA was obtained by sim‐ultaneously using lysozyme ,lysostaphin and chelex100R resin solution ,the obtained Ct value was significantly lower than that of the other components of schizolysis solutions when PCR was used to detect mecA gene of obtained DNA .There was no statistically sig‐nificant difference between adopting the 56 ℃ one‐step method and the 37 ℃ and 56 ℃ two‐step method for conducting MRSA schizolysis(P> 0 .05) ,but the steps were simplified .Conclusion Incubating MRSA in solution containing lysozyme ,lysostaphin , chelex100R resin for 30 min at 56 ℃ is the convenient and efficient schizolysis method to extract DNA ,which can be used immedi‐ately for the next step of PCR and lays the foundation for PCR rapid detection of clinical MRSA infection .

Objective To investigate the drug resistance situation and Staphylococcal cassette chromosome mec(SCCmec) genotypes of methicillin resistant Staphylococcus aureus (M RSA ) strains isolated from Shang-hai Putuo District People′s Hospital in order to provide a theoretical basis for predicting the trend of drug re-sistant bacterial strains and clinical treatment and prevention of MRSA .Methods Three hundreds and eighty clinically isolated MRSA strains in this hospital were collected from January 2012 to December 2016 .The in vitro drug susceptibility test was performed by adopting the broth microdilution method .The SCCmec geno-types were examined by adopting the multiplex polymerase chain reaction .Results All strains were sensitive to linezolid and vancomycin ,the sensitivity rate was 100 .0％ ;the resistance rates to rifampicin and cotrimox-azole were lower ,which were 5 .0％ and 7 .6％ respectively ;but the strains were highly resistant to erythromy-cin ,levofloxacin and tetracycline ,with the resistance rate of 100 .0％ ,94 .2％ ,93 .4％ and 90 .0％ .The resist-ance rate to penicillin was 100 .0％ .Among 380 strains of MRSA ,there were 281 strains(73 .9％ ) of SCCmecⅡ ,59 strains (15 .5％ ) of SCCmecⅢand 5 strains (1 .3％ ) of SCCmecⅣa ,other 35 strains(9 .2％ ) of MRSA could not be classified .Conclusion M RSA strains isolated in the Shanghai Putuo District People′s Hospital are mainly the type SCCmecⅡ ,w hich has the multi-drug resistant characteristics ,and the drug resistance spec-trum of different SCCmec genotypes is different .

Bronchial asthma(asthma)is a common clinical respiratory disease mediated by multiple cells and components,and neutrophils,as one of the classic innate immune cells,play an important role in the pathogenesis of asthma.In the inflammatory state,neutrophils release neutrophils extracellular trap(NET)into the periphery through the NETosis procedure.Although NET is a beneficial antibacterial defense structure,its excessive accumulation can trigger different response mechanisms that can also adversely affect the lungs and exacerbate asthma.Based on the basic principles of traditional Chinese medicine constitution,exploring the important role of NET in the pathogenesis of asthma,it holds that the life process theory body-spirit composition theory,endowment inheritance theory,and environmental constraint theory are related to the gradual formation of NET and different stimuli affect the heterogeneity of NET,thereby mediating different biological effects during the onset of asthma.Targeting NET and/or its components may become a new treatment strategy on asthma.

Electroacupuncture treatment of primary insomnia is widely used and with confirmed efficacy. The factors influencing the therapeutic effect of electroacupuncture include waveform selection, treatment frequency, stimulation intensity, stimulation time and acupoint selection. The mechanism of electroacupuncture in treating this disease mainly includes regulating the levels of excitatory neurotransmitters, γ-aminobutyric acid, melatonin and interleukin cytokines.

Objective:To report a case of Kufor-Rakeb syndrome caused by novel ATP13A2 mutation, collect the cases related to ATP13A2 gene mutation published in recent years, summarize the clinical manifestations of the disease, and broaden the clinical diagnostic thinking. Methods:The clinical manifestations of a newly diagnosed patient with Kufor-Rakeb syndrome caused by ATP13A2 gene mutation admitted to Zhongda Hospital, Southeast University on November 26, 2021, were summarized. The related cases of ATP13A2 mutation published from January 2000 to December 2021 were searched through the PubMed and CNKI databases using the keywords "ATP13A2" and "Parkinson′s disease". The onset age, clinical symptoms, family history, genetic testing, and levodopa responsiveness results of the patients were collected. Results:The patient is a 52-year-old female with the main clinical symptoms of static tremor and bradykinesia. Physical examination showed a gear like increase in muscle tension in the right upper limb, involuntary shaking of the right hand and slow movement. She had good responsiveness to levodopa, and the magnetic resonance imaging and susceptibility weighted imaging of the head showed a lack of clear observation of bilateral black matter swallowtail sign. Whole exome sequencing showed that mutations c.3010A>G (p.S1004G) and c.1195+5G>A (splice) were found in the ATP13A2 gene, both of which were not reported. The c.3010A>G (p.S1004G) mutation originated from the mother, and the c.1195+5G>A (splice) mutation originated from the father. In the retrospective literature review, a total of 10 cases were collected, with onset ages ranging from 18 months to 24 years. Among them, 4/10 patients′ parents married close relatives, and the clinical manifestations were mainly motor symptoms of Parkinson′s disease. In addition, 5/10 patients had cognitive dysfunction, and 3/10 patients had mental symptoms. And demonstrations of most patients′ magnetic resonance imaging were normal in the early stage of the disease, and as the disease progressed, some patients′ imaging results showed specific changes, such as whole brain atrophy and changes in the corpus callosum. Meanwhile, 8/10 patients showed good responsiveness to levodopa. Conclusions:Kufor-Rakeb syndrome is a special type of adolescent levodopa responsive Parkinson′s disease caused by ATP13A2 mutation, which is an autosomal recessive disorder. In addition to motor symptoms such as static tremor and bradykinesia, its clinical manifestations may also be accompanied by non motor symptoms such as cognitive and psychiatric disorders. The disease responds well to treatment with levodopa.

Objective To analyze the clinical outcomes of early invasive fungal disease(IFD)in patients after allogenetic hematopoietic stem cell transplantation(allo-HCST)with metagenomic next-generation sequencing(mNGS).Methods A retrospective analysis was conducted on patients undergoing allo-HCST in our Bone Marrow Transplantation Center between July 2021 and October 2022.These patients experienced one of the following conditions within 100 d after transplantation:① Patients with persistent fever and negative blood culture after empiric antimicrobial therapy for 72 h or longer;② Hyperpyrexia of unknown origin occurred again after effective anti-infection in the past;③ Symptoms in lower respiratory tract associated with lung lesions on CT scan,and empiric anti-infective therapy was ineffective.Peripheral blood or bronchoscopic alveolar lavage fluid were tested with mNGS,and overall survival(OS)and non-relapse mortality(NRM)were analyzed.Results There were 60 patients enrolled in this study.For the peripheral blood samples of 47 cases and bronchoalveolar lavage fluid samples of 13 cases,mNGS found that 19 cases were negative to pathogens,30 cases were non-fungal positive,and 11 case were fungal positive,including 3 cases of aspergillus,5 cases of mucor,2 cases of Candida tropicalis,and 1 case of Trichosporon asahii.Of the 11 patients with fungal positive,8 achieved complete remission after antifungal therapy according to the mNGS results.The 1-year OS and NRM of the 60 patients were 70.0％(95％CI:64.1％～75.9％)and 20.0％(95％CI:11.9％～32.5％),respectively,while those of the fungal infection patients were 54.5％(95％CI:49.5％～69.5％)and 36.4％(95％ CI:15.5％～70.3％),respectively.No significant differences were seen in 1-year OS(P=0.487)and 1-year NRM(P=0.358)among the negative,fungal infection and non-fungal infection patients,neither OS(P=0.238)and NRM(P=0.154)between the fungal infection and the non-fungal infection patients.Conclusion mNGS can rapidly diagnose the early IFD after allo-HSCT,which is helpful for timely and effective treatment and improves the prognosis of patients.

Objective:To achieve rapid and accurate detection of trace uranium in drinking water by analyzing the factors influencing the accuracy of uranium measurement in drinking water using ultraviolet fluorescence method and by evaluating the uncertainty in measurement.Methods:The influence of acidity, Fe 3+ and Mn 2+ contents on the analitical result were studied to optimize the measurement conditions. The accuracy of the measurement method was verified in 7 laboratories. By studying the errors introduced in the process of standard preparation, sample pretreatment and measurement, the sources of uncertainty were analyzed and the uncertainty was synthesized. Results:At pH 1-11 in aqueous solution, the linear regression coefficient of the standard curve was greater than 0.995, which was in line with the linear measurement range of the instrument. At pH 12 or so, the linear regression coefficient was 0.761, which could not meet the measurement requirements. At pH<3 or pH>10, the increase in fluorescence count was lower, which might increase the measurement error. At Fe 3+ concentration ≥15 mg/L, a large deviation occurred in measurement value that could affect seriously measurement result. At Mn 2+ concentration ≥ 1.6 mg/L, the sample produced white precipitation, which could affect the measurement accuracy. Three spiked water samples with different concentrations were determined in 8 laboratories. Each water sample was measured six times in parallel. The relative standard uncertainty of the result were 6.42×10 -2, 4.48×10 -2 and 5.26×10 -2 μg/L, and the expanded uncertainties were 0.03, 0.06 and 0.12 μg/L( k=2), respectively. Conclusions:The optimum conditions for the determination of uranium in water using this method pH were in samples 3-10, the concentration of Fe 3+ less than 15 mg/L, and the concentration of Mn 2+ less than or equal to 1.6 mg/L. The main sources of uncertainty in the measurement of uranium in water using ultraviolet fluorescence method arise from the repeated measurement error and the volume of added standard solution.

Objective:To study the influence of source thickness and counting efficiency calibration on the measurement of gross alpha activity in water.Methods:241Am and natural uranium reference materials were spiked in drinking water to prepare source on a planchet with different thickness, for counting alpha activity on the planchet. Results:The effective thickness measured by spiking 241Am or uranium standard solution in water sample was consistent with the empirical value of 4 mg/cm 2. The alpha counting rate was in a linear increase trend from 2A-5A mg/cm 2 and was basically stable and no longer increase when thickness was higher than 10A mg/cm 2 (A was area of planchet). The result calculated by effective thickness method and thick source method were in good agreement when thickness was 10A mg/cm 2. Conclusions:In order to reduce the deviation of gross alpha counting rate caused by the source thickness and counting efficiency calibration, the source thickness is recommended to be 10A mg/cm 2.

Objective : To investigate possible functional disorders of central auditory processing and language in school-age children with cleft palate through an assessment of the characteristics of the P300 and N400 event-related potentials (ERPs). Methods : This study included 28 school-age children with cleft palate, aged 6 to 12 years, and 30 children without cleft palate as a control group. The P300 and N400 ERPs were selected as indexes of the central auditory processing and language functions of children in both groups. The data were statistically compared between the two groups. Results : Compared with the controls, the children with cleft palate showed a significantly prolonged P300 latency (331.73 ± 14.94 ms vs. 348.64 ± 14.66 ms, P < 0.05) and a significantly decreased P300 amplitude (13.47 ± 2.24 μV vs. 12.07 ± 2.46 μV, P < 0.05). Similarly, the N400 latency of children with cleft palate was significantly prolonged compared to that of controls (431.07 ± 17.90 ms vs. 408.23 ± 18.04 ms, P < 0.05), and the N400 amplitude was significantly decreased compared to that of controls (13.75 ± 2.12 μV vs. 15.17 ± 2.34 μV, P < 0.05). Conclusion School-age children with cleft palate may have central auditory processing disorders and language dysfunctions.

ObjectiveTo investigate the potential mechanism of action of the Qufeng Tongqiao Cough-relieving Decoction （祛风通窍止咳方， QTCD） in the treatment of upper airway cough syndrome （UACS）. MethodsTwenty-four guinea pigs were randomly divided into blank group， model group， traditional Chinese medicine （TCM） group， and inhibitor group， with six guinea pigs in each group. Except for the blank group， guinea pigs were sensitized with ovalbumin and aluminum hydroxide via intraperitoneal injection， followed by ovalbumin nasal drops combined with smoke exposure to establish the UACS model. After modeling， the TCM group was administered QTCD 0.9 g/（100 g·d） by gavage， the inhibitor group received the transient receptor potential vanilloid receptor 4 （TRPV4） inhibitor GSK2193874 1 mmol/L， 5 min by nebulisation， and the blank group and model group were given 2 ml/（100 g·d） normal saline by gavage once daily. After 7 days of treatment， a cough provocation test was performed using 0.4 mol/L citric acid. The levels of IgE in serum and inflammatory cytokines， including interleukin-6 （IL-6）， interleukin-8 （IL-8） in serum， bronchoalveolar lavage fluid （BALF）， and nasal lavage fluid （NLF） were detected by enzyme-linked immunosorbent assay （ELISA）. Histopathological changes in lung and nasal mucosal tissues were observed by hematoxylin-eosin （HE） staining. Immunohistochemistry was used to detect the protein levels of TRPV4， substance P （SP）， and calcitonin gene-related peptide （CGRP） in lung and nasal mucosal tissues. Real-time polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of TRPV4， SP， and CGRP in lung tissues. ResultsHE staining showed significant structural damage and infiltration of inflammatory cells in the lung and nasal mucosal tissues in the model group， while the TCM group and inhibitor group showed improved pathological changes. Compared with the blank group， the model group showed increased cough frequency， serum IgE level， and IL-6 and IL-8 levels in serum， BALF， and NLF. The protein levels of TRPV4， SP， and CGRP in lung and nasal mucosal tissues and their mRNA expression were elevated （P＜0.05 or P＜0.01）. Compared with the model group， the TCM group and inhibitor group showed reduced cough frequency， serum IgE level， and TRPV4 and SP mRNA expression in lung tissues. The TCM group showed reduced IL-6 and IL-8 levels in serum， BALF， and NLF， and reduced TRPV4 and CGRP protein levels in lung and nasal mucosal tissues. The inhibitor group showed reduced IL-6 and IL-8 levels in serum， BALF， and NLF， reduced IL-6 in BALF， reduced IL-8 in NLF， and decreased TRPV4， SP， and CGRP protein levels in lung tissues and SP and CGRP protein levels in nasal mucosal tissues （P＜0.05 or P＜0.01）. Compared with the TCM group， the inhibitor group had increased serum IgE， IL-6， and IL-8 levels， increased IL-6 level in BALF， and increased IL-8 levle in NLF， but decreased SP protein level in lung tissues and increased TRPV4 and SP mRNA expression in lung tissues （P＜0.01）. ConclusionQTCD effectively reduces cough frequency in the UACS guinea pig model. Its mechanism may involve inhibiting the activation of the TRPV4 pathway， improving airway neurogenic inflammation， alleviating inflammatory responses， and reducing cough hypersensitivity.