Objective To explore the radiosensitizatian of paclitaxel in human lung adenocareinoma cells. Methods A human lung adenocarcinoma cell line A973 was used in this study. The cytotoxicity of paclitaxel was investigated by using clonngenic assay to define the IC10,IC50 and IC90. The cells received either radiation(with different doses) alone or paclitaxel administrated before and after irradiation. Cell survival fractions were determined by clonogenic assay. Single hit multi-target model was used to determine survival curve parameters. Flow cytometry was performed to analyze the cell cycle distribution. Results The IC10, IC50 and IC90 of paclitaxel in A973 cells were 0.5,2.6 and 8.7 nmol/L,respectively. According to Do,Dq and SF2 value,the sensitivity enhancement ratio(SER) of IC10 was 0.97,1.01 and 2.00 when paclitaxel was added before irradiation, and 0.97,1.02 and 1.02 when after irradiation ; The SER of IC50 was 1.06,129.00 and 2.61 when paclitaxel was added before irradiation, and 0.94,220. O0 and 2.14 when after irradiation ;The SER of IC90 were 1.00,220. 00 and 2.09 when paclitaxel was added before irradiation,and 0.98,220.00 and 2.09 when after irradiation. The IC10 of paclitaxei failed to increase G2+M arrest of A973 cells.The maximal G2+M accumulation was reached at 2 h and 18 h after IC50 and IC90 of paclitaxel treatment,respectively. Conclusions Paclitaxel is able to enhance the radiation sensitivity of A973 cells. Sequence of treatment is not associated with radiosensitivity. Moderate and high dose of paclitaxel combined with low-dose radiation can produce the best effect of radiosensitiation.

ObjectiveTo investigate the effect of Glutamine (Gln) on the expression of PCNA in intestinal tissue of neo-natal rats with necrotizing enterocolitis (NEC), and to explore the protective mechanism of Gln in intestinal mucosa.Methods Forty-eight neonatal rats at the age of 48 hours were selected, and divided into 4 groups, control group, Gln group, NEC group, NECGln group. Each group had 12 rats. Control group were fed mice milk substitutes; Gln group were fed mice milk substitutes mixed with Gln; NEC group were fed mice milk substitutes and had cold/ hypoxia exposure twice a day for 3 days; NECGln group were exposed to cold stress, hypoxia and treated with Gln mixed in the milk. The expression of PCNA was detected using immunohistochemical method.Results Compared with control group were and Gln group, the general condition was worse, and the weight was decreased in NEC and NECGln group. The inifltrated inlfammatory cells, congestion, edema, intrinsic layer separation were observed in intestinal mucosa in NEC and NECGln group. The intestinal villus was lost in severe in NEC and NECGln group. The PCNA index was 34.17±5.78, 34.42±5.38, 15.00±1.94, 30.67±3.14 in control, Gln, NEC and NECGln group respectively, with signiifcant difference between each groups (H=24.32,P=0.000). The expression of PCNA in NEC group was lower than that in normal, Gln, and NECGln group (P<0.008). The expression of PCNA had no signiifcant difference among normal, Gln, and NECGln group (P>0.008).Conclusions The expression of PCNA in intestinal mucosa was decreased in NEC rats. Gln supplement could raise the expression of PCNA in intestinal mucosa of NEC rats, and accelerate the speed of intestinal mucosa repair.

Aged ; Benzylisoquinolines ; therapeutic use ; Bronchoalveolar Lavage ; Female ; Humans ; Male ; Middle Aged ; Oxidative Stress ; Phytotherapy ; Pneumoconiosis ; metabolism ; therapy ; Quality of Life ; Treatment Outcome

Objective To investigate the value of multi-b value DWI combined with PSA evaluation on the effect of endocrinotherapy for prostate cancer (PCa), and to study the correlation between the signal intensity(SI), apparent diffusion coefficient(ADC) with PSA.Methods Forty patients with PCa diagnosed by pathology or biopsy were tested for serum PSA levels before and after endocrine therapy.All patients underwent DWI at b-value of 300,800 and 1 000 s/mm2 by using a GE Signa HDx 3.0T MRI scanner.The serum PSA,the value of SI and ADC at different b values before and after endocrine therapy,and their correlation with PSA were observed and measured, respectively.Results (1)The serum PSA before and after endocrine therapy were (35.63±20.91) ng/mL and (5.98±3.84) ng/mL, respectively.The difference was statistically significant (P<0.05).(2)On the condition of different b values, ADC values were (1.06±0.06)×10-3 mm2/s, (0.83±0.04)×10-3 mm2/s, (0.73±0.03)×10-3 mm2/s, respectively before endocrine therapy, and (1.33±0.09)×10-3 mm2/s, (1.16±0.09)×10-3 mm2/s, (1.10±0.08)×10-3 mm2/s, respectively after endocrine therapy.There were significant difference between the ADC values before endocrine therapy and those after endocrine therapy (P<0.05).SI were 809.09±79.06, 417.42±26.65 and 327.42±25.26 in 40 patients with Pca before the endocrine therapy, and 780.10±78.52,409.33±33.93 and 320.00±31.10 after endocrine therapy.The difference was not statistically significant (P>0.05).(3)Pearsoncorrelation analysis showed that ADC value was negatively correlated with serum PSA (r300=-0.58,P<0.01;r800=-0.60,P<0.01;r1 000=-0.66,P<0.01) before endocrine therapy.ADC value was also negatively correlated with serum PSA (r300=-0.55,P<0.01;r800=-0.52,P<0.01;r1 000=-0.61,P<0.01) after endocrine therapy.In addition,SI was negatively correlated with serum PSA (r300=-0.09,P<0.01;r800=-0.18,P<0.01;r1 000=-0.28,P<0.01) before the treatment, as well.Conclusion ADC value at different b values increased after endocrine therapy in Pca, and is negatively correlated with serum PSA.The correlation is the most significant at b value of 1 000 s/mm2.The combination of DWI and serum PSA could be used to monitor and evaluate the effect of endocrinotherapy for Pca, and better guide the clinical treatment of Pca.

ObjectiveTo establish a droplet digital PCR (ddPCR) method for detecting hepatitis B virus (HBV) covalently closed circular DNA (cccDNA). MethodsHBV cccDNA standard substance was constructed, and HBV cccDNA primers and probes were designed based on the structural differences between HBV cccDNA and relaxed circular DNA (rcDNA). HBV plasmid was amplified to obtain HBV cccDNA standard substance, and a ddPCR detection method was established with the standard substance after gradient dilution as the template for HBV cccDNA detection; the limit of detection and repeatability of this method were analyzed. Liver tissue samples were collected from 20 patients who attended Beijing YouAn Hospital, Capital Medical University, from June 2017 to October 2020, all of whom were diagnosed with HBV infection, and DNA of the samples was extracted and digested with plasmid-safe ATP-dependent DNA enzyme to obtain HBV cccDNA template; the ddPCR detection method was evaluated in clinical samples and was compared with the quantitative real-time PCR (qPCR) detection method. The chi-square test was used for comparison of categorical data between the two groups. ResultsThe HBV cccDNA detection method based on ddPCR was established, which accurately detected HBV cccDNA in standard substance after gradient dilution, with a limit of detection of 1 copy/μl, and the coefficients of variation of 1×103, 1×102, and 1×101 copies/μl standard substances were 441%, 3.98%, and 5.09%, respectively. HBV cccDNA was detected in the samples of 20 patients with HBV infection; the ddPCR detection method detected HBV cccDNA in 17 patients, with a positive rate of 85%, while the qPCR detection method detected HBV cccDNA in 11 patients, with a positive rate of 55%, and there was a significant difference between the two methods (χ2=4.286, P=0038). ConclusionThe established ddPCR method for detecting HBV cccDNA has a low limit of detection and good repeatability, which provides an effective tool for further clinical detection.

Objective To explore the use of queuing theory model combined with the dynamic allocation of nursing staff in the management of blood collection room in outpatient. Methods Queuing theory model combined with dynamic allocation of nursing staff were applied to scientifically allocate nurses in blood collection room. Three months before and after allocation, nursing staff, inspection personnel and patients were investigated through satisfaction questionnaire and the management effect was evaluated. Results Patients′satisfaction degree toward blood collection staff was increased from 80% to 95%; inspection personnel′s satisfaction degree toward blood collection staff was increased from 62.5% to 87.5%;nurses′satisfaction degree toward themselves was increased from 70% to 95% (χ2=20.571,13.333,5.22;P<0.05) . Patients′waiting time before blood collection was shorten to 20 min from 30 min. Conclusions The application of queuing theory model combined with dynamic allocation of nursing personnel can improve the rational use of blood collection personnel in outpatient and nursing quality, and it also can improve the satisfaction of inspection personnel to nurses, of patients and nurses to themselves.

Objective To investigate the endoplasmic reticulum stress(ERS)role in the course of liver failure induced by severe hepatitis B virus(HBV)infection and its related mechanism.Methods Liver tissue samples and clinical data [chronic hepatitis B patients(12 cases,chronic hepatitis B group),hepatic failure induced by severe hepatitis B virus(12 cases,severe hepatitis B virus liver failure group),and normal subjects(8 cases,control group)] were collected from the Beijing You'an Hospital affiliated to Capital Medical University between 2009 to 2011.Statistical analysis was performed on the clinical indicators of each group.The structure of endoplasmic reticulum in liver tissue was observed by transmission electron microscopy.Western blot and qRT-PCR were used to detect the expression of endoplasmic reticulum stress and apoptosis-related factors,including glucose-regulated protein(Grp),and C/EBP homologous protein(CHOP).Frozen sections of liver tissues were prepared for immunofluorescence test.All data were expressed as mean±standard deviation.LSD-t test was used to compare the results between groups.A p value<0.05 was considered as statistically significant.Results Transmission electron microscopy showed that the morphological structure of the endoplasmic reticulum was damaged in both groups(chronic hepatitis B and liver failure induced by severe hepatitis B virus),and liver failure induced by severe hepatitis B virus group was more critical.Western blot and qRT-PCR showed that Grp78,Grp94 and Caspase-4 were highly expressed in normal group and chronic hepatitis B group,and the relative protein expressions were 1.20±0.13 and 0.78±0.11,0.90±0.06 and 0.11±0.01,0.15±0.02 and 0.22±0.04,respectively.The expression of protein was weakened in liver failure induced by severe hepatitis B virus group(relative protein expression was 0.01±0,0.01±0,and 0.11±0.02,respectively).There was a statistically significant difference between the two groups(P<0.05).The expression of CHOP was consistent with the results of immunofluorescence,and increased with the stressing of injury.Conclusion During the course of severe hepatitis B infection,dysregulated endoplasmic reticulum stress activated mild stress in chronic hepatitis B group,while severe stress in hepatic failure induced by severe hepatitis B virus group.Therefore,endoplasmic reticulum stress plays an important and complex role in the pathogenesis of hepatic failure induced by severe hepatitis B virus.

Objective:To explore the expectations and values of patients with malocclusion on participation in shared decision-making of orthognathic surgical protocols, and to provide references for further development of clinical shared decision-making models.Methods:Based on the expected value theory and descriptive qualitative research methods, using purposive sampling, 13 patients with malocclusion in the Ninth People's Hospital of Shanghai Jiao tong University School of Medicine from May to August in 2021 were selected for semi-structured interviews. The interview data were sorted, classified and refined by traditional content analysis.Results:Two themes were extracted: patients' ability beliefs about their ability to participate in shared decision making for orthognathic surgery(decision support ability, psychological coping ability and environmental adaptability), and task values for shared decision making for orthognathic surgery(interest value, acquisition value).Conclusions:Low level of patients′ ability beliefs in shared decision-making, active physician guidance facilitates patient participation, but the depth of patient participation is influenced by factors such as information support, cultural climate, and physical space for shared decision making. It is suggested that the magnetic role of physicians should be actively played, the shared decision-making team should be strengthened, at the same time, hospital manager should enhance shared decision making propaganda to increase the acceptance and participation of patients in shared decision making so as to improve the quality of shared decision-making.

Deep venipuncture catheterization is a routine and basic operation in the treatment of critically ill patients, and it is the most effective way to quickly correct the shock. Clinical B-ultrasound guided deep vein catheters can improve the success rate of puncture, but in the process of operation, the short axis needs to be replaced by the long axis. In the replacement process, the stability of the novice is insufficient, the positioning is difficult, and the operation time is too long. If only short axis puncture is used, it is impossible to know whether the current position of the puncture needle, and the puncture may be too deep and stray into the artery. The accuracy of the 45 degree angle of the injection point requires a very experienced operator. In view of the above shortcomings, doctors in the department of critical care medicine of Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine designed a B-ultrasound puncture equipment, which has obtained the National Invention Patent of China (ZL 2016 1 0571557.X). The device is composed of B-ultrasound probe fixing frame, sliding scale plate, simulation slide rule, puncture needle, sliding device. By sliding device the angle of the pinhole channel, it is conducive to the accurate positioning of the puncture target, optimizing the operation procedure, improving the puncture speed and accuracy, effectively reducing the occurrence of puncture complications, ensuring patient safety, reducing unnecessary waste of human and material resources. It can reduce the workload of medical staff and is worthy of clinical practice.
Humans ; Catheterization, Central Venous/methods* ; Ultrasonography, Interventional/methods* ; Ultrasonography ; Punctures/methods* ; Needles

Objective:To establish a rapid and accurate method for the detection of Klebsiella pneumoniae carbapenemase (KPC) carbapenemase gene based on recombinase aided amplification (RAA)-CRISPR-Cas13a (CRISPR-Cas13a) technology. Methods:Twenty-five clinical isolates of carbapenem-resistant Klebsiella pneumoniae (CRKP) and five carbapenem-sensitive Klebsiella pneumoniae (CSKP) strains preserved in 2020-2021 in Beijing Chuiyangliu Hospital were randomly collected, and the total DNA samples of the strains was extracted. RAA primers specific for KPC DNA and CRISPR RNA (crRNA) were designed to establish a rapid and accurate method for the detection of KPC carbapenemase gene based on RAA-CRISPR-Cas13a technology. The method was evaluated by plasmids and clinical sample strains, and the detection was also performed by Quantitative real-time PCR (qPCR) method to compare the detection rate and consistency of the two methods. Results:The RAA-CRISPR-Cas13a method can detect KPC plasmids and samples with a sensitivity of 1 copy/μl, which is higher than that of qPCR (10 1 copies/μl). Among the 30 clinical strains (including 25 CRKP strains and 5 CSKP strains), 23 strains were detected to carry KPC gene by both RAA-CRISPR-Cas13a method and qPCR method, and 7 strains were not detected with KPC gene. The detection rate of KPC gene in the 25 CRKP strains was 92% (23/25). The positive coincidence rate of the two methods was 100% (23/23). Conclusions:This study combined RAA amplification technology with CRISPR-Cas13a technology to establish a rapid and accurate method for detecting KPC carbapenemase gene. The method is useful for accurate screening of KPC carbapenemase-producing strains. It has a wide application prospect in drug resistance monitoring and infection control.