Objective To investigate the prevalence of Echinococcus infections in small rodents around human residential areas in Yushu City, Qinghai Province in 2023, so as to provide insights into precision echinococcosis control. Methods One or two quadrats, each measuring 50 m × 50 m, were randomly assigned in Shanglaxiu Township and Longbao Township, Yushu City, Qinghai Province on June 2023, respectively, and 300 plate-type mouse traps, each measuring 12.0 cm × 6.5 cm, were assigned in each quadrat. Small rodents were captured during the period between 10 : 00 and 18 : 00 each day for 4 days. Then, all captured small rodents were identified and dissected, and liver specimens with suspected Echinococcus infections were subjected to pathological examinations. The Echinococcus cytochrome c oxidase 1 (cox1) gene was amplified using PCR assay, and the sequence of the amplified product was aligned to that was recorded in the GenBank to characterize the parasite species. In addition, a phylogenetic tree of Echinococcus was generated based on the cox1 gene sequence using the neighbor-joining method. Results A total of 236 small rodents were captured in Shanglaxiu and Longbao townships, Yushu City, including 65 Qinghai voles and 51 plateau pikas in Shanglaxiu Township, and 62 Qinghai voles and 58 plateau pikas in Longbao Township, and there was no significant difference in the constituent ratio of small rodents between the two townships (χ² = 0.294, P > 0.05). Seven plateau pikas and 12 Qinghai voles were suspected to be infected with Echinococcus by dissection, and pathological examinations showed unclear structure of hepatic lobules and disordered hepatocyte arrangement in livers of small rodents suspected of Echinococcus infections. PCR assay identified E. shiquicus DNA in 7 Qinghai voles, which were all captured from Shanglaxiu Township. Phylogenetic analysis showed that the cox1 gene sequence of Echinococcus in small rodents was highly homologous to the E. shiquicus cox1 gene sequence reported previously. Conclusion Plateau pika and Qinghai vole were predominant small rodents around human residential areas in Yushu City, Qinghai Province in 2023, and E. shiquicus infection was detected in Qinghai voles.

Novel coronavirus Omicron variant infection can cause severe illness and even death in certain populations. Omicron variant infection may lead to systemic inflammatory response, coagulation disorder, multi-organ dysfunction and other pathophysiological changes, which are different from other Novel coronavirus variants to a certain extent, so therapeutic strategies should not be the same. The National Medical Center for Major Public Health Events invited experts in fields of infectious diseases, respiratory medicine, intensive care, pediatrics and fever clinic to develop this quick guideline based on the current best evidence and extensive clinical practices. This quick guideline aims to standardize the diagnosis and treatment of novel coronavirus Omicron infection, and to improve the disease management abilities of clinicians.

Objective:To analyze the clinical characteristics of necrotizing otitis externa(NOE) and to discuss its clinical staging and treatment.Methods:A retrospective case series study was conducted in the Otorhinolaryngology Head and Neck Surgery Center of Beijing Friendship Hospital. Patients hospitalized in our department with diagnosis of NOE between June 2015 to December 2021 were included. There were 23 patients (23 affected ears), with 19 males and 4 females, aged 32-85 years (the median age was 68 years old). We analyzed and summarized the main symptoms and signs, laboratory examinations, imaging examinations, treatments and prognosis of the patients, and established a database for data collection and sorting.Results:All the 23 patients had various complications. There were 19 cases of type 2 diabetes, 5 cases of anemia, 3 cases of hypoalbuminemia, 2 cases of chronic renal insufficiency, and 1 case of uremia. The main symptoms on admission were otalgia, otorrhea, hearing loss, angular deviation, limited mouth opening, dysphagia and hoarseness. Physical examination showed swelling of the external ear canal (EEC), granulation tissue of the EEC, thin purulent secretion of the EEC, facial paralysis, restricted mouth opening, and decreased vocal cord movement. Etiological results showed 15 cases of Pseudomonas aeruginosa, 5 cases of Staphylococci and 3 cases of Aspergillus fumigatus. Imaging mainly showed abnormal soft tissue shadows of the EEC and middle ear, and temporomandibular joint, parapharyngeal space, skull base, intracranial and other adjacent structures were involved according to the different invasion directions of the lesions. According to the characteristics of these patients, we summarized a diagnostic criteria and clinical staging. All patients received systematic anti-infection, surgery and symptomatic support treatment. Until January 30, 2022, 4 patients died, including 1 case of extensive intracranial infection caused by NOE, and 3 cases of other systemic diseases after the affected ears were recovered and discharged from hospital. The other 19 patients′ ears were cured, the symptoms of otalgia and otorrhea disappeared, and the epithelization of the surgical cavity was good. Conclusion:The clinical manifestations of NOE lack specificity, and the diagnosis should be made according to the medical history, physical examination and auxiliary examination. According to different clinical stages, corresponding treatment measures should be formulated. The treatment is based on multidisciplinary comprehensive treatment, mainly surgical treatment. Early intervention has a good prognosis, otherwise, multiple-group of cranial nerve injury or intracranial infection implies poor prognosis.

Objective To explore the effects of two methods of air-impact on clearing the subglottic secretion in patients with intubation.Methods A simple random sampling method was used to select 106 patients underwent mechanical ventilation through oral tracheal intubation in intensive medicine department from September 2016 to October 2017.The recruited patients were divided into two groups by the random number table,53 patients in Group A were treated with breath-holding key of a ventilator,combined with air-bag inflation and deflation,and 53 patients in Group B were treated with simple breathing apparatus combined with manual technique to clear subglottic secretion.The incidence of ventilator-associated pneumonia(VAP),amount of cleared subglottic secretions,difference of vital signs before and after operation,number of coughing,in vitro training time and operation time were compared between groups.Results The intention-to-treat ana]ysis(ITT) showed that the incidence of VAP in Group A and B were 7.55％ and 5.66％,the per-protocol analysis(PP) showed that the incidence of VAP in Group A and B were 3.92％ and 3.85％,and there was no significant difference between groups(P＞0.05);the amount of cleared subglottic secretions in two groups were (8.31±0.82) ml,(7.97±1.12)ml,and there was no significant difference (P＞ 0.05);but vital signs before and after operation,number of coughing,in vitro training time and operation time in Group A were lower than those in Group B,and the differences were statistically significant(P＜0.05).Conclusion Two methods of air-impact can both effectively reduce the incidence of VAP,but using breath-holding key of a ventilator combined with airbag inflation and deflation has less influence on vital signs,which patients can better tolerate and medical staff can master and cooperate more easily.

60 curved root canals of permanent teeth with angles of curvature ranging from 15°to 40°(Schneider's methodology)were prepared using the instruments of Hyflex CM(HC) and ProTaper(PT) Universal respectively(n=30).Using standardized pre-and post-instrumentation paralleling periapical radiographs,canal curvature was determined by image analysis software and the clinical shaping effect of Hyflex CM and ProTaper rotary NiTi files were compared.The canal curvature in group HC and PT decreased by 4.54°±3.25° and 5.63°±3.84° respectively(between pre-and post-treatment in both groups,P<0.05;between groups,P>0.05).Hyflex CM can meet the clinical necessity for the instrumentation of curved root canals.

Objective The inhibitor of differentiation 3 (Id3) is an important transcriptional regulation factor, which participates in tumorigenesis, cell proliferation, and cell apoptosis.β-catenin, as a central molecule of the Wnt signaling pathway, is critical for tumor development.This study aimed to evaluate the expressions of these two molecules and the regulatory effect of Id3 on β-catenin in different tumor cells.Methods Total RNA was extracted using the Trizol Reagent.The relative mRNA expression levels of Id3 and β-catenin in tumor cells were detected by quantitative real-timePCR(qRT-PCR).The recombinant eukaryotic expression vector pEGFP/Id3 with the human Id3 gene was transfected into A549, A549/ DDP and SW-480 cells using the non-liposome-mediated method.The protein expressions of Id3 and β-catenin were determined by Western blot.Results The expression of Id3 was significantly lower in the colorectal cancer cell lines SW-480 and HT-29 than in A549 and other tumor cells (P<0.05), but remarkably higher in nasopharyngeal carcinoma CNE and 5-8F cells than in other tumor cells (P<0.05).The expression of β-catenin was the highest in SW-480in comparison withother malignant tumor cells(P<0.05), and the second highest was in gastric cancer AGS and colorectal cancer HT-29 cell lines, but low in H446, A549, SPC-A-1, A549/DDP, and SK-MES-1 cell lines and extremely low or almost absent in CNE and 5-8F cells (P<0.05).After transfected with pEGFP/Id3, the cells showed a decreased volume, wrinkled membrane and absent refraction under the fluorescence microscope, which, however, were not observed in most of the cells transfected with the empty vector pEGFP.Compared with the control, the Id3/pEGFP group showed remarkably increased expressions of Id3 mRNA in the A549, A549/DDI, and SW-480 cells (1.24±0.12 vs 193.12±2.80, 1.09±0.11 vs 188.30±2.60, and 0.92±0.29 vs 19.08±0.59, P<0.01), and the expression of β-catenin was significantly down-regulated in the transfected SW-480 cells with an overexpression of Id3 (0.98±0.05 vs 0.32±0.03, P<0.01), but exhibited no statistically significant differences from those in the transfected A549 and A549/DDP cells (0.98±0.07 vs 1.04±0.08 and 0.98±0.05 vs 0.32±0.03, P>0.05).Western blot showed the same results.Conclusion The expression levels of Id3 and β-catenin vary in different tumor cell lines.Anabnormally high level of β-catenin is an important risk factor for colorectal cancer, and the down-regulatedexpression of β-catenin after eogenous transfection of Id3 may provide some new ideas for target gene therapies of colorectal cancer.

Objective Rheumatoid arthritis (RA) is a typical type Ⅲ hypersensitivity with a large number of immune complexes (IC) and complement deposits in the synovial tissue , but its specific pathogenesis is not yet clear.This article was to explore the expression of the antigenic profile of serum ICs in RA.Methods ICs were isolated from the serum of 55 patients with RA (41 cases of anti-CCP antibody [+] and 14 cases of anti-CCP antibody [-]), 41 with systemic lupus erythematosus (SLE), and another 41 healthy controls by polyethylene glycol (PEG) precipitation, separated by immunoprecipitation, digested with trypsin in gel, and then subjected to mass spectrometry for identification.The levels of total proteins were compared among different groups using Vennny 2.1.0.The protein expression was considered to be up-regulated when the total protein level of the RA group was >2 times and down-regulated when it was <0.5 times that of the control.Further functional analysis was performed on the differential proteins in RA using the STRING software.Results Totally, 277 proteins were identified in the serum ICs of the RA patients, including 162 in the anti-CCP (+) and 248 in the anti-CCP (-) RA group.Compared with the SLE and healthy control groups, only 129 proteins were found in the RA patients, including 38 in the anti-CCP (+), 109 in the anti-CCP (-) RA group, and 18 in both the two groups.Among the proteins identified in the RA patients and healthy controls, 2 and 11 were up-regulated while 17 and 21 down-regulated in the anti-CCP (+) and anti-CCP (-) RA group, respectively.Conclusion More differentially expressed proteins were identified in the anti-CCP (-) than in the anti-CCP (+) RA patients.The identification of differentially expressed proteins provides a new idea and direction for the investigation of the pathogenesis and new biomarkers of RA.

Objective To scan protein expression profile of immune complexes (ICs) derived from the synovial tissue of the patients with rheumatoid arthritis (RA) based on liquid chromatography-tandem mass spectrometry (LC-MS).Methods The samples of synovial fluid were obtained from knee joints of the patients with RA and osteoarthritis (OA) used as control during therapeutic arthrocentesis in knee jiont at the Department of Orthopedics of Jinling Hospital,School of Medicine,Nanjing University.The protein expression profile of ICs was identified by enrichment strategy based on immunoprecipitation and LC-MS analysis.The value of fraction of total (FOT) was used to estimate protein abundance and screen the up-and down-regulated proteins.The function enrichment,interaction network and signal pathway of differential proteins were analyzed using softwares David and String.Results A total of 511 and 526 protein spots in ICs of RA and OA patients were identified respectively.Among them,170 proteins existed only in RA group.45 and 85 proteins in RA group were statistically up-and down-expressed compared with controls.Conclusion HSP90AA1,HSP70,HLAG,Thioredoxin,Annexin A2 and vitronectin may be involved in the pathogenesis of RA through different paths and possible to become promising diagnostic indicators or new therapeutic targets for RA.

Objective:To establish a method to determine the content of chlorogenic acid and rutin in nettle to assess the quality of nettle.Methods:An HPLC method was used with the following chromatographic conditions:CAPCELL PAK C18 column (250 mm × 4.6 mm, 5 μm) was used with the mixture of 0.4%phosphoric acid and methanol as the mobile phase with gradient elution , the de-tection wavelength was set at 340 nm, the flow rate was 1.0 ml· min-1 and the sample size was 10μl.Results:Chlorogenic acid with-in the range of 2.360-23.600μg · ml-1 showed a good linear relationship (r=0.999 9), and rutin within the range of 6.208-62.080μg · ml-1 showed a good linear relationship (r=0.999 9).The recovery of rutin and chlorogenic acid was 99.20% and 100.40%with RSD of 1.2%and 1.1%(n=6), respectively.Conclusion:The method is fast, effective, simple, accurate and reproducible , which can be used to quantitatively analyze chlorogenic acid and rutin in nettle .

OBJECTIVE: To explore the survival rate of neural stem cells (NSCs) from fetal rat and its biological properties after cryopreservation and thawing. METHOD: Different generations NSCs (the first generation, the third generation and the sixth generation) from fetal rat which cultivated with serum-free medium in vitro were cryopreserved in the cryogen, which were neurosphere culture medium with 10% BSA and 7.5% DMSO (without neural growth factor). The cryopreserved cells were resuscitated at 1st week, 4th week, 8th week, 12th week and 16th week respectively. The survival rate of cells were calculated and the cells were incubated and differentiated again. RESULT: Different time of cryopreservation, different generations did not affect NSCs survival (P > 0.05) after cryopreservation. The survival rate of NSCs was from 60% to 70% after resuscitated, which were differentiated into neurons and astrocytes in 10% embryonic bovine serum (without growth factor). CONCLUSION NSCs were successfully cryopreserved, resuscitated and recultured, which would create bases for the experimental study on the selected-date application of NSCs transplantation into cochlea for treating sensorineural deafness.
Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Proliferation ; Cell Survival ; Cells, Cultured ; Cryopreservation ; Female ; Fetus ; cytology ; Neural Stem Cells ; cytology ; Neurons ; cytology ; Pregnancy ; Rats ; Rats, Sprague-Dawley