OBJECTIVE To investigate the effects of ligustilide on chemotherapy resistance of cervical cancer cells based on Hippo-Yes-associated protein （YAP） signaling pathway. METHODS Human cervical cancer cisplatin-resistant cells HeLa/DDP were divided into control group， cisplatin group （10 μmol/L cisplatin）， cisplatin+ligustilide low-， medium- and high-concentration groups （10 μmol/L cisplatin+25， 50， 100 μmol/L ligustilide）. The proliferation， apoptosis， migration and invasion of HeLa/DDP cells were all detected in each group. The mRNA expressions of YAP and transcriptional coactivator with PDZ binding motif （TAZ） as well as the protein expressions of YAP， TAZ， matrix metalloproteinase 2 （MMP2）， Ki67， cleaved-caspase-3 and caspase-3 were determined in HeLa/DDP cells. RESULTS Compared with control group， the inhibitory rate， apoptotic rate and cleaved- caspase-3/caspase-3 of cisplatin group were increased significantly； scratch healing rate， the number of invasive cells， the mRNA expressions of YAP and TAZ， and the protein expressions of YAP， TAZ， MMP2 and Ki67 were decreased significantly in cisplatin group （P＜0.05）. Compared with cisplatin group， the inhibitory rate of cell proliferation， apoptotic rate and cleaved-caspase-3/ caspase-3 were further increased in cisplatin+ligustilide low-， medium- and high-concentration groups， while scratch healing rate， the number of invasive cells， the mRNA expressions of YAP and TAZ， and the protein expressions of YAP， TAZ， MMP2 and Ki67 were further decreased， in a dose-dependent manner （P＜0.05）. CONCLUSIONS Ligustilide can increase the sensitivity of drug-resistant cervical cancer cells to cisplatin by inhibiting Hippo-YAP signaling pathway.

Objective To investigate the clinical features and genetic characteristics of patients with late-onset Pompe disease(LOPD).Methods A total of 13 patients diagnosed with LOPD in the First Affilia-ted Hospital of Zhejiang University School of Medicine from September 2020 to December 2023 were selected,and all patients were subjected to clinical investigation,GAA activity detection and GAA gene testing.Results Among the 13 patients,7 were males and 6 were females;5 were family patients and 8 were sporadic patients;and the median age of onset was 17 years(8-52 years),the median age of presentation was 24 years(10-52 years),and the median age of diagnosis was 31 years(14-58 years).In terms of the first symptoms,10 pa-tients presented with limb weakness and 3 patients presented with dyspnea.The average serum creatine kinase level was 552 U/L(55-1084 U/L),and the serum creatine kinase level was normal in one patient.All pa-tients had scoliosis and different degrees of restrictive ventilatory dysfunction.Neuroelectrophysiological exami-nations of 9 patients showed myogenic damage,and 8 of them had muscle tonic discharge.The mean value of GAA activity was 0.3 μmol/(L·h)[0.17-0.5 μmol/(L·h)].A total of 13 mutations were detected in GAA gene,and the most common mutation was c.2238G＞C(p.W746C).There were five new variant sites:c.543del(p.F181Lfs*40),c.839_840insCC(p.R281Pfs*34),c.1800_1823del(p.S601_R608del),c.2296T＞C(p.Y766H)and c.995C＞A(p.S332*).Conclusions LOPD is a rare disease that tends to delay diagnosis.Proximal limb weakness,decreased respiratory function,mild-to-moderate elevation of creatine kinase,scoliosis,and clinical inferior tonic discharge on electromyography are high-risk images of LOPD.c.2238G＞C(p.W746C)is a hotspot mutation,and the discovery of five new mutations enriches the GAA gene mutations lineage.

Objective The present study was conducted to identify the Vibrio cholera type and to analyze its antibiotic resistance in an epidemic of cholera in Haiyan County in 2018, which would provide the references for prevention and control of cholera. Methods Stool samples of the patient and his close contacts as well as the food and environmental samples were collected for identification of the type of Vibrio cholerae and the toxin gene. The resistance of identified Vibrio cholerae to 20 different common antibiotics were tested. Results A total of 176 samples were collected, including 101 stool samples from the case and his close contacts, 35 environmental samples and 40 food samples. Among those samples, only one strain of V. cholerae, O139, was isolated from the patient's first feces sample. It was detected as a toxin gene of ctxA positive by real-time fluorescence PCR. Antibiotic resistance test showed that the strain was sensitive to norfloxacin, levofloxacin, ciprofloxacin, cefotaxime, cephalothin, ampicillin, and amoxicillin. It was 100% resistant to tetracycline, doxycycline, neomycin, kanamycin, streptomycin, and rifampicin. Conclusion V. cholerae O139 strain with ctxA is detected in an epidemic of cholera. Norfloxacin, levofluoxacin and some other antibiotics could be used for clinical treatment and prevention. It should pay attention to this strain of V. cholera regarding the multiple drug resistance and the change of antibiotic resistance.

Breast cancer brain metastases (BCBMs) are one of the most difficult malignancies to treat due to the intracranial location and multifocal growth. Chemotherapy and molecular targeted therapy are extremely ineffective for BCBMs due to the inept brain accumulation because of the formidable blood‒brain barrier (BBB). Accumulation studies prove that low density lipoprotein receptor-related protein 1 (LRP1) is promising target for BBB transcytosis. However, as the primary clearance receptor for amyloid beta and tissue plasminogen activator, LRP1 at abluminal side of BBB can clear LRP1-targeting therapeutics. Matrix metalloproteinase-1 (MMP1) is highly enriched in metastatic niche to promote growth of BCBMs. Herein, it is reported that nanoparticles (NPs-K-s-A) tethered with MMP1-sensitive fusion peptide containing HER2-targeting K and LRP1-targeting angiopep-2 (A), can surmount the BBB and escape LRP1-mediated clearance in metastatic niche. NPs-K-s-A revealed infinitely superior brain accumulation to angiopep-2-decorated NPs-A in BCBMs bearing mice, while comparable brain accumulation in normal mice. The delivered doxorubicin and lapatinib synergistically inhibit BCBMs growth and prolongs survival of mice bearing BCBMs. Due to the efficient BBB penetration, special and remarkable clearance escape, and facilitated therapeutic outcome, the fusion peptide-based drug delivery strategy may serve as a potential approach for clinical management of BCBMs.

Objective Analyze the application status and existed problems of clinical biobank in China,propose possible application mechanisms for clinical biobank.Methods Through questionnaire survey and case analysis combined with relevant literature reports from home and abroad,conduct qualitative analysis to understand the application status and problems of clinical biobank.Results With the rapid development of biobank,its application rate was far from expected.The construction of clinical database lags behind in China will affect the application of samples.The lag of application mechanism will affect the opening and application of the biobank.Conclusions At the beginning of construction,the clinical biobank should take full account of what and how resources can be used,and establish tailored application mechanism.More attention should be paid to the possible benefit of biobank construction.

Venous leg ulcer (VLU) is a disease of venous hypertension leading to open skin lesions of lower extremities, which is the terminal stage of chronic venous insufficiency. Considering the high prevalence and recurrence rate, VLU restrain daily activities, impair mental health and even contribute to social isolation of patients. Currently, surgery and compression therapy are wildly used to relieve symptoms and eliminate the etiological factors, however, ulcers remain unhealed even after these treatments in many cases. In this article, we review the clinical progress of treatment for VLU, including the debridement, antibiotics, ulcer dressing, negative pressure wound therapy and skin graft, to provide reference on the therapeutic strategy for different ulcer conditions.

Objective To explore the diagnosis and treatment of empyema in children. Method The clinical data of empyema in 49 children were reviewed and analyzed. Results In the 49 cases (25 males and 24 females) aged 4.7±3.4 years, the common symptoms were fever, shortness of breath and coughing. There were 11 cases of positive blood culture, 17 cases of positive pleural fluid culture, 3 cases of positive blood and pleural fluid culture. Streptococcus pneumoniae was the most common pathogenic bacteria. All 49 patients were given systemic antibiotics and closed thoracic drainage. In addition, 26 cases were treated with urokinase and intrapleural fibrinolytic therapy and 6 cases were treated surgically. The prognosis was good and there was no death. Conclusion Pneumococcal infection is most common in children with empyema. Systemic antibiotics plus closed thoracic drainage and urokinase are effective, and some require surgical treatment.

Objective Based on Total Quality Management (TQM) theory,our study aims to analyze the clinical Bio-Bank overall quality management implications,basic characteristics,principles,and management mechanisms,and provide theoretical basis for the clinical Bio-Bank quality construction.Methods Using theoretical and literature research methods,Bio-Bank overall quality management qualitative analysis was conducted,putting forward a framework of Bio-Bank comprehensive quality management.Results Biological sample overall quality management was defined theoretically including its connotation,concepts and basic characteristics.We also put forward an application principle and basic operation method at the application level.Conclusions Total Quality Management (TQM) is applied to the clinical Bio-Bank construction,from where,the scientific and unique management content can effectively optimize the Bio-Bank management regulation and standardization of the sample operation process in the PDCA cycle,which is critical to improve the quality of clinical Bio-Bank.

Objective To explore the effects of flipped classroom combined with problem-based learning (PBL) teaching mode on the self-learning ability in nursing students. Methods From September 2015 to June 2016, students from 6 classes of nursing major of Grade 2013 in Kangda College of Nanjing Medical University were selected by convenience sampling method. Class 1 to 3 were assigned as the experimental group, and Class 4 to 6 were assigned as the control group. In the control group, traditional teaching mode was applied, while in the experimental group, flipped classroom combined with PBL teaching mode was adopted. The self-learning ability was compared between the two groups. Results After intervention, the self-efficacy, learning control, the total score of motivation subscale, learning help, learning planning, learning summary, learning management and the total score of the strategy subscale of the experimental group were all higher than those of the control group, and the differences were statistically significant (P< 0.05). Conclusions The flipped classroom combined with PBL mode can improve students' self-learning ability, which is worthy of promotion.

Objective To investigate the effects of abdominal breathing training on sleep disorders in elderly patients with chronic heart failure. Methods Total of 100 patients with chronic heart failure complain of sleeping disorders and Pittsburgh Sleep Quality Index (PSQI)>7 points were assigned into two groups by random digits table method, 50 cases in each group. The observation group and the control group were nursed in the same way except that abdominal breathing was adapted to the observation group. Sleep status, heart rate, blood pressure, SpO2 and brain natriuretic peptide (BNP) were evaluated before training, one week and eight weeks after training respectively. Statistics was used to analyze the differences between two groups. Results After training one week, the sleep status of the observation group was ameliorated, but without significant difference compared to the control group (P>0.05). And after training eight weeks, the PSQI, BNP and heart rate were (9.21 ± 6.38) points, (193.78 ± 152.16) μg/L, (63.5 ± 10.8) times/min in the observation group, and (12.92 ± 0.33) points, (417.55 ± 262.47) μg/L, (70.7 ± 8.5) times/min in the control group, and there was significant differences between 2 groups (t=3.627, 2.041, 2.767, all P < 0.05), while the blood pressure, SpO2 did not change obviously(P>0.05). Conclusions Abdominal breathing training could ameliorate sleep status in elderly patients with chronic heart failure.