Abstract：Objective To investigate the morphological features of the Pneumocystis jirovecii, in order to facilitate early detection and rapid diagnosis of this rare pathogen from a morphology point of view by laboratory technicians. By analyzing the laboratory features and application value of different pathogen detection methods in the diagnosis of Pneumocystis jirovecii pneumonia, we aim to provide the most reliable diagnostic basis for rapid diagnosis of Pneumocystis jirovecii pneumonia.Methods A retrospective analysis was conducted on the test results of bronchoalveolar lavage fluid samples from a comprehensive hospital in Zhangqiu District, Jinan City, Shandong Province, and a hospital in Changde City from April 2022 to October 2022. Five confirmed cases of Pneumocystis jirovecii pneumonia were detected. Its clinical manifestations, laboratory results, and morphological characteristics of pathogens under different stains were analyzed to discuss the advantages and disadvantages of different detection methods. Results Cytological examination of bronchoalveolar lavage fluid found the trophozoites and cysts of Pneumocystis jirovecii by Wright's-Giemsa staining in 4 cases (80%), and the cysts of Pneumocystis jirovecii by Silver hexamine staining in 4 cases (80%), while the metagenomic next-generation sequencing confirmed all the 5 positive results. All 5 patients had different degrees of reduction in the absolute count of peripheral blood lymphocytes, and the serum lactic dehydrogenase and (1-3)-β-D-Glucan were increased. Among the 5 patients in this study, 4 were treated with sulfamethoxazole combined with caspofungin, and 1 was treated with sulfamethoxazole. Three patients were cured and discharged from hospital after treatment, but two died. Conclusions The method of Wright's-Giemsa staining for the cytological examination of bronchoalveolar lavage fluid to find Pneumocystis jirovecii has the unique and irreplaceable advantages as silver staining. Metagenomic next-generation sequencing can further increase the positive detection rate of Pneumocystis jirovecii. The combination of cytological examination of bronchoalveolar lavage fluid with metagenomic nextgeneration sequencing is a powerful diagnostic method for rapid diagnosis of Pneumocystis jirovecii pneumonia, which can diagnose accurately and reduce missed diagnosis.

Objective:To evaluate the effect of botulinum neurotoxin A (BoNT/A) on prevention of chronic migraine (CM) in mice and explore the potential mechanism.Methods:Twenty-four male C57BL/6 mice were randomly divided into control group, nitroglycerin (NTG) group, and BoNT/A+NTG group ( n=8). Mice in the latter two groups were intraperitoneally injected with 10 mg/kg NTG on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments to establish CM models. Mice in the BoNT/A+NTG group were injected with 0.18 U/100 μL BoNT/A one h before the first injection of NTG. Mice in the control group were injected with the same dose of normal saline. Basal mechanical withdrawal threshold (MWT) and evoked MWT 2 h after NTG in the facial and hindpaw regions on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments were evaluated by von Frey filament test. The motor function of mice 2 h after NTG injection was tested by rotarod test on the 1 st, 3 rd, 5 th, 7 th and 9 th d of experiments. On 9 th d of experiments, the mice were sacrified; the calcitonin gene-related peptide (CGRP), synaptosomal-associated protein 25 (SNAP25), glial fibrillary acidic protein (GFAP) and TRP channel protein expressions in the trigeminal ganglia (TG) and trigeminal nucleus caudalis (TNC), and NOD-like receptor protein 3 (NLRP3) inflammatory factor pathway-related protein expressions in TNC were detected by Western blotting; real-time quantitative PCR (RT-qPCR) was used to detect the NLRP3 inflammatory factor pathway-related mRNA expressions in TNC. The CGRP expression in TNC was detected by immunofluorescent staining. Results:(1) As compared with the control group, the NTG group had significantly decreased basal facial MWT on the 7 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased basal facial MWT on the 7 th and 9 th d of experiments ( P<0.05). As compared with the control group, the NTG group had significantly decreased evoked facial MWT on the 5 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased evoked facial MWT on the 5 th and 9 th d of experiments ( P<0.05). As compared with the control group, the NTG group had significantly decreased basal and evoked MWT in the hindpaw regions on the 3 rd, 5 th, 7 th and 9 th d of experiments ( P<0.05); as compared with the NTG group, the BoNT/A+NTG group had significantly increased basal and evoked MWT in the hindpaw regions on the 3 rd, 5 th, 7 th and 9 th d of experiments ( P<0.05). (2) There was no significant difference in running time on rotarod among the three groups ( P>0.05). (3)Western blotting results showed that as compared with those in the control group, the CGRP and SNAP25 protein expressions were significantly increased in TG of the NTG group ( P<0.05); and those in the BoNT/A+NTG group were significantly decreased as compared with those in the NTG group ( P<0.05). As compared with those in the control group, the CGRP and NLRP3 protein expressions were significantly increased in TNC of NTG group ( P<0.05); and those in the BoNT/A+NTG group were significantly decreased as compared with those in the NTG group ( P<0.05). (4)RT-qPCR results showed that as compared with that in the control group, the IL-1β mRNA expression in TNC of the NTG group was significantly increased ( P<0.05), and that in the BoNT/A prevention group was statistically decreased as compared with that in the NTG group ( P<0.05). (5) Immunofluorescent staining results showed that as compared with that in the control group, the CGRP expression in TNC of the NTG group was significantly increased, and that in the BoNT/A+NTG group was significantly decreased as compared with that in the NTG group ( P<0.05). Conclusion:BoNT/A can reduce the SNAP25 expression in TG, reduce the CGRP release in TG and TNC, and prevent CM onset; BoNT/A can regulate NLRP3 level in TNC.

Objective:To investigate the clinical efficacy and safety of 90Sr- 90Y β-ray low dose applicator, topical timolol maleate, and their combination in the treatment of superficial infantile hemangioma (IH). Methods:From May 14, 2013 to April 11, 2017, 400 children (126 males, 274 females, age 5.3(3.9, 7.1) months) with superficial IH in Department of Nuclear Medicine, the First People′s Hospital of Foshan were prospectively enrolled. All patients were randomly divided into 4 treatment groups according to the proportion of 1∶1∶1∶1 by the method of random number table: topical timolol maleate (group A, control group), 90Sr- 90Y β-ray low dose applicator (group B), single course applicator combined with timolol (group C), and multi-course applicator combined with timolol (group D). Lesions were followed up to the 104 th week (W104). Cure rate of W104 was considered as primary end point. Efficacy and safety of different treatment were compared. Kruskal-Wallis rank sum test, Mann-Whitney U test, and logistic regression analysis were used for statistical analysis. Results:Totally, 438 lesions in 400 cases were included in this prospective study. There was no significant difference in baseline characteristics among 4 groups ( χ2 values: 1.709-11.616, H values: 3.681-7.653, all P>0.05). As of W104, 11 lesions (2.51%, 11/438) were lost follow-up, 32 lesions (7.31%, 32/438) were with early withdrawal, 357 lesions (81.51%, 357/438) were cured, 15 lesions (3.42%, 15/438) were with residual, 23 lesions (5.25%, 23/438) were with rebound growth, and no serious adverse events occurred in the 4 groups. Multivariate analysis showed that lesions thickness (<3 mm vs ≥ 3 mm, odd ratio ( OR)=16.689, 95% CI: 7.908-35.223; χ2=54.555, P<0.001) and treatment (considering group A as reference category, OR (95% CI) of group B, C and D were 16.842(6.179-45.901), 4.801(2.167-10.638) and 39.127(10.468-146.243), respectively; χ2=47.663, P<0.001) were independent factors affecting the cure rate of W104. 90Sr- 90Y low-dose fractionation radiotherapy was significantly better than topical timolol maleate ( OR=16.842, 95% CI: 6.179-45.901), and the combination with timolol could significantly reduce the cumulative absorbed dose of radiotherapy (group D vs B: 16(8, 16) vs 16(16, 24) Gy; z=-4.947, P<0.001). Conclusion:90Sr- 90Y low dose applicator therapy is superior to topical timolol maleate for superficial IH, and the combination with timolol could significantly reduce the cumulative absorbed dose of applicator.

Objective? To investigate the effect of the cardiac rehabilitation program based Omaha system on patients after percutaneous coronary intervention (PCI). Methods? A convenient sampling method was used to select 127 patients from the Department of Cardiology of Wuxi Second People's Hospital from October 2017 to April 2018. According to the patient's hospitalization number, they were randomly divided into two groups, including 64 in the experimental group and 63 in the control group. During the intervention, 2 patients were lost in the experimental group and 1 patient was lost in the control group. There were 62 patients in each group to complete the study. Patients in the control group received routine care, while patients in the experimental group received cardiac rehabilitation based on the Omaha system. The effect of the intervention was measured by 6 min exercise test, secondary prevention knowledge and behavior questionnaire for post-PCI patients. Results? After the intervention, The 6-minute exercise distance was (439.28±67.94) m in the experimental group, and it was (315.62±45.18) m in the control group. The exercise endurance of the experimental group was higher than that of the control group, and the difference was statistically significant (t=11.934, P＜ 0.01). The secondary prevention knowledge and behavior scores of the experimental group were (87.21±12.46) and (84.27±14.58), which were higher than those of the control group (71.30±8.12) and (65.94±8.97). The differences were statistically significant (t=8.423, 8.431; P ＜0.01). Conclusions? Cardiac rehabilitation programs based Omaha system can improve exercise endurance and secondary prevention behaviors of patients after PCI and its application should be promoted.

Objective To explore the application of timing theory in home care of patients with acute coronary syndrome.Methods A total of 90 patients with acute coronary syndrome admitted to the Second People's Hospital of Wuxi in Jiangsu Province from February 2016 to February 2017 were selected, and randomly divided into control group and study group, with 45 cases each. The control group was given routine nursing intervention, while the study group was given home-based care intervention based on timing theory. The quality of life and negative emotions were compared between the two groups, using Self-Rating Anxiety Scale (SAS), Self-Rating Depression Scale (SDS), Seattle Angina Questionnaire (SAQ) and Athens Insomnia Scale (AIS).Results The scores of SAS and SDS of the control group after intervention were (61.7±2.0) and (52.4±1.3) respectively, which were both higher than those of the observation group, and the differences were statistically significant (t=27.295,23.129;P<0.001). After 3 months of nursing intervention, the quality of life of patients in both groups has been improved. SAQ scores in different division were as follow: limitation of physical activity (64.1±10.9), angina attack (89.7±12.0), stable angina pectoris (91.5±7.2), disease cognition (88.5±9.2) and satisfaction of treatment (91.6±10.8). Those scores were all higher than those of the control group. The differences were statistically significant (P<0.05). The score of sleeping time of the study group was (1.8±0.6), and that of the control group was (1.5±0.5). The difference in the sleeping quality between the two groups was statistically significant (t=2.577,P<0.05).Conclusions Home care intervention based on timing theory for patients with acute coronary syndrome can improve the quality of life and treatment effect, reduce anxiety and depression, and is worth popularizing and applying.

Objective: To explore influence of nursing strategies based on evidence based medical theory on negative emotion and daily behavior in aged patients with coronary heart disease (CHD).Methods: A total of 88 CHD patients, who were treated in our department from Oct 2012 to Mar 2015, were selected.According to random number table, they were randomly and equally divided into routine nursing group and evidence based nursing group (received evidence based nursing based on routine nursing group).Unhealthy lifestyles, scores of self-rating anxiety scale (SAS) and self-rating depression scale (SDS), satisfaction for nursing and incidence rate of major adverse cardiovascular events (MACE) were measured and compared between two groups before and four months after intervention.Results: Compared with routine nursing group after four-week intervention, there were significant reductions in percentages of smoking (40.9% vs.22.7%), drinking (54.5% vs.38.6%), high-fat diet (43.2% vs.29.5%), physical inactivity (45.5% vs.25.0%) and overweight (52.3% vs.29.5%), scores of SAS [(50.4±3.3) scores vs.(45.8±3.2) scores] and SDS [(53.4±4.7) scores vs.(48.0±4.2) scores], and incidence rate of MACE (25.0% vs.11.4%), and significant rise in satisfaction for nursing (86.4% vs.97.7%) in evidence based nursing group (except for the P of overweight was <0.01, others were P<0.05 all).Conclusion: Evidence based nursing mode can improve unhealthy lifestyle and relieve negative emotional status in aged patients with coronary heart disease, which is worth extending in clinic.

Objective:To establish the fingerprint analysis method for the root of Rosa laevigata Michx from different regions by UPLC. Methods:The column was ACQUITY UPLC? Phenyl(2.1 mm × 100 mm,1.7 μm). The mobile phase consisted of methanol-water with gradient elution. The flow rate was 0. 2 ml·min-1 , the detection wavelength was 210 nm, the column temperature was 30℃, and the injection volume was 3 μl. Results:The fingerprint consisted of 15 common peaks. The range of similarity for twelve bat-ches of the root of R. laevigata Michx was 0. 489-0. 942. And the reference fingerprint of the root of R. laevigata Michx was estab-lished by UPLC. Conclusion:The fingerprint method is simple and reproducible, which can provide basis for the quality control and the medicinal resources exploration.

Objective To investigate the effects of "one to one" tutorial system of two-way selection model in the standardized training of new nurses.Methods Using purposive sampling, a total of 25 new nurses recruited in July 2014 were selected as the control group,and they received traditional teaching mode. Another 25 new nurses recruited in July 2015 were selected as the observation group,and they received "one to one" tutorial system of two-way selection model. The theoretical and operating scores,general self-efficacy and career self-efficacy of nurses were compared between two groups after one year's training.Results The sores of basic theory,basic operation,professional theory and bedside practice in the observation group were (89.99±5.76), (90.08±2.56),(88.39±6.71) and (92.74±4.30),which were significantly higher than (85.61±6.27),(87.99±3.27),(84.30±5.74) and (89.22±3.55) in the control group (t=2.105,2.296,2.591,2.543;P<0.05). There was no significant difference between two groups in the scores of general self-efficacy,educational requirements and job duties before the training (P>0.05). After one year's training,the scores of general self-efficacy,educational requirements and job duties in the observation group were (31.48±3.12),(15.24±2.16) and (15.64±2.48),which were significantly higher than (26.96±3.54),(11.66±2.54) and (12.40±2.50) in the control group (t=3.901,4.177 and 4.825;P<0.01).Conclusions "One to one" tutorial system of two-way selection model in the standardized training of new nurses could make new nurses change from students to qualified nurses faster and it is worth to be expanded.

Objective To explore the clinical effect of laparoscopic electrocoagulating hook'method in treatment of rectal cancer.Methods The retrospective cohort study was adopted.The clinical data of 237 patients with rectal cancer who were admitted to the Second People's Hospital of Guangxi Zhuang Autonomous Region from June 2004 to June 2009 were collected.Among 237 patients undergoing radical resection of rectal cancer,120 using laparoscopic electrocoagulating hook'method were allocated into the laparoscopic electrocoagulating hook'group and 117 using laparoscopic ultrasound knife's method were allocated into the laparoscopic ultrasound knife's group.Observation indicators included:(1)treatment situations:operation time,volume of intraoperative blood loss,number of lymph node dissected,time to anal exsufflation,time for out-of-bed activity,duration of hospital stay,treatment expenses and occurrence of complications(anastomotic leakage,intestinal obstruction,postoperative dysuresia and sexual dysfunction),(2)follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect tumor recurrence and metastasis up to December 18,2014.The colonoscopy,chest X-ray and abdominal computed tomography(CT)were applied to suspected patients with local or distal tumor recurrence.Measurement data with normal distribution were represented asx±s and comparison between groups was analyzed using t test.Count data were analyzed using chi-square test or Fisher exact probability.The survival curve was drawn by Kaplan-Meier method,and the survival rate was analyzed using the Log-rank test.Results(1)Treatment situations:all the patients underwent successful laparoscopic surgery,without conversion to open surgery.Operation time,number of lymph node dissected,time to anal exsufflation,time for out-of-bed activity and duration of hospital stay were respectively(156±33)minutes,18±2,(1.5±0.6)days,(1.5±0.5)days,(7.5±2.5)days in the laparoscopic electrocoagulating hook'group and(175±21)minutes,16±4,(1.6±0.5)days,(1.4±0.5)days,(8.0±1.8)days in the laparoscopic ultrasound knife's group,with no statistically significant difference in the above indexes between the 2 groups(t=25.324,20.117,21.477,1.617,1.770,P＞0.05).Volume of intraoperative blood loss and treatment expenses were respectively(116±12)mL,(2.4±0.3)×104 yuan in the laparoscopic electrocoagulating hook'group and(149±25)mL,(2.8±0.2)×104 yuan in the laparoscopic ultrasound knife's group,with statistically significant differences in the above indexes between the 2 groups(t=212.740,211.071,P＜0.05).Anastomotic leakage,intestinal obstruction and dysuresia or sexual dysfunction were respectively detected in 6,3,0 in the laparoscopic electrocoagulating hook'group and 7,3,5 in the laparoscopic ultrasound knife's group,with no statistically significant difference in the anastomotic leakage and intestinal obstruction between the 2 groups(x2=0.110,P＞0.05)and with a statistically significant difference in the dysuresia or sexual dysfunction between the 2 groups(P＜0.05).All the patients received postoperative chemotherapy of FOLFOX regimens for 6 cycles.(2)Followup situations:226 patients were followed up for a median time of 52 months(range,11-121 months),and 11 patients lost to follow-up were included in the study as censored data,including 5 in the laparoscopic electrocoagulating hook'group and 6 in the laparoscopic ultrasound knife's group.During the follow-up,67 patients were dead,including 58 died of tumor recurrence and metastasis,4 died of cerebral hemorrhage,3 died of myocardial infarction and 2 died of renal failure.The overall survival rate and overall survival time were 70.36％and 51 months.The overall local tumor recurrence rate,distal tumor recurrence rate and 5-year survival rate were 8.33％(10/120),25.83％(31/120),71.7％in the laparoscopic electrocoagulating hook'group and 7.69％(9/117),29.06％(34/117),72.4％in the laparoscopic ultrasound knife's group,with no statistically significant difference between the 2 groups(x2=0.033,0.310,0.002,P＞0.05).Conclusion The laparoscopic electrocoagulating hook'method in treatment of rectal cancer is safe and effective compared with traditional laparoscopic surgery,and it could also reduce the incidence of autonomic nerve-related complications and treatment expenses.

Objective To understand the probability of premature mortality caused by non-communicable diseases (NCD) in Beijing from 2010 to 2015.Methods The data of deaths from Beijing vital registration system were used to analyze age-standardized NCD mortality rates,proportion of NCD deaths in age group ＜70 years and premature NCD mortality.Results NCD deaths accounted for 90％ of the total deaths in Beijing.Age-standardized NCD death rate decreased from 332.43/100 000 in 2010 to 280.02/100 000 in 2015.Meanwhile,the probability of deaths from four NCDs between in age group 30-70 years (premature NCD mortality) decreased from 12.81％ to 11.11％ in Beijing.The premature mortality of four NCDs in men was 14.63％,and compared with 2015 baseline,it would decrease by 16.4％ to reach 2025 goal (12.23％),and the probability of premature mortality four NCDs in women was 7.54％,and compared with 2015 baseline,it would decrease by 6.8％ to reach the 2025 goal (7.03％).People living in suburb areas had a higher probability of premature NCD mortality (13.67％) than those living in urban areas (9.72％) in Beijing in 2015.Conclusions The premature NCD mortality was much higher in men than in women in Beijing.More attention should be paid to the control of risk factors for NCD premature deaths,especially in men and in suburb residents,to reach the 25 × 25 goal.