OBJECTIVE:To prepare anti-tumor drug H6(lactone compound)polymeric micelles,and to investigate its in vitro anti-tumor effects. METHODS:Using mPEG2000-PCL4000 as carrier,H6/mPEG2000-PCL4000 micelles were prepared. Using particle size, PDI and 48 h whether to produce precipitation as indexes,feeding ratio,H6 concentration,volume ratio of organic solvent were screened. The encapsulation efficiency and drug-loading amount of micelle were all detected. MTT assay was used to detect the tox-icity of micelles and H6 solution to human non-small cell lung cancer cell A549 and human lung cancer cell H460. RESULTS:The screened formulation was as follows as feeding ratio of 1∶25,H6 concentration of 2 mg/mL,the ratio of ethanol to chloroform of 1∶1(V/V). The parameters of prepared H6/mPEG2000-PCL4000 micelles were as follows as particle size of(40.74±0.116 3)nm,PDI of(0.101±0.006),encapsulation efficiency of(94.87±0.016 3)%,drug-loading amount of(7.07±0.001 5)%(n=3). IC50 of mi-celles and H6 solution to A549 cell were 15.62 and 12.57 nmol/L;IC50 of micelles and H6 solution to H460 cell were 27.68 and 15.19 nmol/L. CONCLUSIONS:H6/mPEG2000-PCL4000 micelles are prepared successfully and show in vitro anti-tumor effects.

Objective To construct the recombinant rAAV2-hGAD65 vector and detect its function both in vitro and in vivo. Methods The cDNA of human glutamic acid decarboxylase 2 (hGAD65) gene, which was one of gamma-aminobutyric acid (GABA) synthetase, cloned by the method of RT-PCR, was subcloned into the adeno-associated virus (AAV) vector and formed the recombinant vector of AAV-hGAD65 (rAAV2-hGAD65). The recombinant vector was packaged by the AAV Helper-Free System and its titer was detected. The primary fibroblast, cultured from the rat lung, was transfected by the rAAV2-hGAD65. The expression of the hGAD65 in fibroblast was detected by immunohistochemical method and the level of GABA in the media was assayed by high performance liquid chromatograph (HPLC). In vivo, the hGAD65 was detected by immunohistochemical method in STN and the concentration of the GABA in the reticular part of substantia nigra (SNr) was assayed by HPLC after the rAAV2-hGAD65 was injected into the subthalamic nucleus (STN) by the stereotaxic method. Results The sequence of hGAD65 cDNA was in accordance with that in the Genebank. The amino acid sequence of hGAD65 had no mutation and the titer of rAAV2-hGAD65 was reached 4.5 ×10~(11) per milliliter. The efficiency of infection to the rat primary firoblasts was 80%, and the concentration of GABA in the media of fibroblasts was (45.66±6.07)nmol/L per liter. In vivo, hGAD65 could be detected in STN, and the concentrateion of the GABA in the SNr was increased from (5.66±1.07)nmol/g to (12.66±2.59)nmol/g.Conclusion The cDNA of hGAD65 was cloned by RT-PCR and the recombinant vector of rAAV2-hGAD65 was constructed. The AAV can infect the primary fibroblast in vitro and the hGAD65 can catalyse the glutamic acic to GABA. In vivo, the concentration of GABA in the SNr was heighten after the rAAV2-hGAD65 was injected into the STN.

Objective To study the clinical and pathological manifestations of microsporidian encephalitis.Methods The clinical findings and the brain pathological features of a patient with microsporidian encephalitis hospitalized in 2004 were studied.Results The onset was subacute or chronic. The body temperature was usually normal or below 37.5℃,but it rose when patient's condition deteriorated and coma appeaxed.The patient had hypoimmunity but without human immunodeficiency virus infection. Multifocal lesions in the whole brain,signs of meningeal irritation and infective myelogram were observed. Rheumatoid factor increased in the early stage and indirect bilirubin,proteins in cerebrospinal fluid(CSF), and immunoglobulin IgG,IgA increased in the middle stage.Cytological examination of CSF showed lymphocyte reaction.Blood routine test showed normal eosinophil granulocyte count.The patient was found to have pleurisy,peritonitis and cystitis.Brain magnetic resonance image(MRI)manifested plaque-like isometric T1 weight image and long T2 weight image signal in white matter of bilateral cerebral hemisphere and cerebella where FLAIR sequence showed hyperintensity.No apparent mass was identified.Contrast- enhanced MRI scan showed patchy and ring-like intensification.The neural system impairments were permanent and not improved after treatment.The pathology of brain tissue showed neuronal degeneration, karyopycnosis and Derivasculitis.The infectious agents were observed in the cytoplasm of neurons.Wister rats had muhiple organ inflammatory reaction 2 weeks after intraperitoneal inoculation of the patient's CSF and a large quantity of pathogens were found in the peritoneal lavage fluid.Conclusions The patient was PAS staining method is useful for detecting the pathogen in neurons and the rate can be raised by animal intraperitoneal cultivation

OBJECTIVE:To establish the detection method for anti-inflammatory compound HB0314 in plasma of rats,and study on its pharmacokinetic characteristics in rats in vivo. METHODS:UPLC-MS/MS was performed on the column of Waters Ac-quity UPLCTM BEH C18 with mobile phase of 0.1% formic acid aqueous solution(A)-methanol(B)by gradient elution(0-2 min, 70%-90% B) at flow rate of 0.25 mL/min,column temperature was 30 ℃,and injection volume was 5 μL. Electrospray ion source was used,capillary voltage was 3 kV,ion source temperature was 150 ℃,desolvation gas temperature was 450 ℃,desol-vent air flow volume was 600 L/h,cone air flow volume was 45 L/h,and the inner standard was tetrahydropalmatine. 12 rats were randomly divided into iv group and ig group,6 in each group. Rats were intravenously injected and intragastrically administrated HB0314 solution 5,10 mg/kg. Sample blood 0.4 mL were taken from the jugular vein blood before administration and after 5,15, 30,60,120,240,360,480,600,720,1440 min of administration to determine the HB0314 plasma concentration. DAS 2.0 software was used to calculate the pharmacokinetic parameters and absolute bioavailability. RESULTS:The linear range of HB0314 was 1-1000 ng/mL(r=0.9955),and the lower limit of quantification was 1 ng/mL. RSDs of extra-day and daytime precision,sta-bility were not higher than 8.45%(n=5);recovery were 68.21%-90.29%(RSD≤11.20%,n=5),and matrix effects were 82.63%-106.90%(RSD≤6.75%,n=5). After intravenous injection and intragastric administration,AUC0-24 h were (270.267 ± 21.164), (252.755 ± 26.169)μg·h/L (n=6);t1/2z were (8.722 ± 2.266),(11.877 ± 4.517) h (n=6);and absolute bioavailability was 56.79%. CONCLUSIONS:The method is rapid,simple,and can be used for the determination of HB0314 content in plasma of rats. HB0314 shows high oral absolute bioavailability in rats in vivo,indicating that post-dosage form design may be considered as oral anti-inflammatory drugs.

Objective Plasma circulating DNA can be em-ployed in place of bone marrow examination for the auxiliary diagnosis of leukemia.This study aimed to explore the clinical application of the plasma DNA level in evaluating the effect of chemotherapy on chronic leukemia. Methods We collected blood samples from 52 patients with chronic myelogenous leukemia (CML) (33 in the chronic phase, 7 in the acceleration phase, and 12 in the blast phase) , 85 with chron-ic lymphocytic leukemia (CLL) (28 with complete remission, 27 with partial remission, and 30 with no remission), 4 patients with hairy cell leukemia (HCL), and 80 healthy subjects.We simultaneously obtained plasma DNA and recombinant plasmid DNA using the BI-LATEST DNA Kit and examined the human β-actin gene and the level of plasmid DNA by real-time quantitative PCR. Results Before chemotherapy, the median value of plasma DNA was 149.46(30.63-496.91)ng/ml in the CML and 101.54(69.10-258.14) ng/ml in the CLL patients, both significantly higher than in the healthy controls (19.05[12.67-25.92]ng/ml) (P<0.01).After chemotherapy, the plasma DNA level of the CML patients was remarkably decreased, but still higher than that of the controls ( P<0.01).The CML patients in the chronic phase showed a markedly higher level of plasma DNA (302.89[93.33-541.52]ng/ml) than those in the blast phase (43.19[23.54-70.03]ng/ml) and acceleration phase (28.11[16.21-92.07]ng/ml) (P<0.05).The CLL patients with CR exhibited a significantly lower level of plasma DNA (24.29[14.64-30.74]ng/ml) than those with PR (106.88 [96.23-143.25]ng/ml) and NR (460.73[284.57-653.38〗ng/ml) (P<0.01), but all dramatically higher than that of the healthy controls (P<0.01) Conclusion The quantification of plasma DNA has a clinical application value in evaluating the effect of chemo-therapy on chronic leukemia.

Objective To detect the changes of the NJ001 specific antigen expression before and after surgery, and evaluate whether the NJ001 specific antigen could be used as a serum biomarker for the diagnosis of pancreatic cancer.Methods With the method of sandwich ELISA, the serum samples from 85 pancreatic cancer patients, 22 pancreatic benign tumor and 40 healthy controls were detected respectively. The results of the NJ001 specific antigen in the serum samples from 85 pancreatic cancer patients were compared with CA19-9 detected by ECLIA.Results The positive rate of NJ001 for the pancreatic cancer group was obviously higher than that for the benign pancreatic tumor and health control groups[50.6%(43/85) vs 18.2%(4/22), χ2 =7.451, P<0.05; 50.6%(43/85) vs 10.0%(4/40), χ2 =19.098, P<0.05].The difference between benign pancreatic tumor group and health control group had no statistical significance[18.2%(4/22) vs 10.0%(4/40),χ2 =0.845, P>0.05].The positive rate in the group of pancreatic cancer before surgery was higher than that after surgery[50.6%(43/85) vs 23.5%(20/85),χ2=13.341, P<0.05].In addition, the results from 85 pancreatic cancer patients showed the specificity of NJ001 specific antigen was up to 87.1%.Although the positive rate of NJ001 specific antigen for pancreatic cancer was lower than that of CA19-9[50.6%(43/85) vs 75.3%(64/85), χ2 =11.121, P<0.05], it was higher when they combined [ 85.9%( 73/85 ) ] .Conclusions It shows high positive rate of NJ001 specific antigen in the patients of pancreatic cancer in this study, which suggests that NJ001 specific antigen might be a potential valuable biomarker for the diagnosis of pancreatic cancer.

Objective:To explore the effect of urapidil combined with nitroglycerin on blood pressure in elderly patients with acute hypertension.Methods:From July 2016 to September 2018, 60 elderly patients with acute hypertension admitted to the First People's Hospital of Yongkang were selected as the observation objects.They were divided into control group and observation group according to random number table method, with 30 cases in each group.The control group was treated with nitroglycerin intravenous drip, while the observation group was treated with urapidil hydrochloride injection by intravenous micropump on the basis of the control group.The total effective rate, diastolic blood pressure, systolic blood pressure and heart rate were compared between the two groups at the time of admission and 60 minutes after treatment.Results:The effective rate of the observation group was 96.67%, which was significantly higher than 76.67% of the control group (χ 2=5.19, P<0.05). There were no statistically significant differences in blood pressure and heart rate between the two groups at the time of admission (all P>0.05). After treatment, the blood pressure of the two groups decreased significantly (all P<0.05), and the decrease levels of the observation group was significantly better than those of the control group, the difference were statistically significant (all P<0.05). The heart rate of the observation group was significantly lower than that of the control group ( P<0.05). The incidence of adverse reactions in the observation group (10.00%) was significantly lower than that in the control group (33.33%), and the difference was statistically significant (χ 2=4.81, P<0.05). Conclusion:Intravenous micropump injection of urapidil hydrochloride injection combined with nitroglycerin has remarkable antihypertensive effect, less adverse reactions and high safety in the treatment of elderly patients with acute hypertension.

Objective To explore the impact of mindfulness on sleep and the mediating role of resilience.Methods A total of 540 medical staff in a three first-class hospital were assessed by five facet mindfulness questionnaire (FFMQ),Pittsburgh sleep quality index(PSQI) and Connor-Davidson resilience scale(CD-RISC).Results The average PSQI scores of medical staff was (6.67±3.20),of which the total score was equal or above 8 accounting for 37.04％.The positive rate of each factor of PSQI (factor score ≥2) was 51.67％ for daytime function,37.22％ for sleeping time,and 24.07％ for subjective sleep quality.The total score of PSQI was (6.67±3.20),the score of FFMQ was (119.55±9.90),and the score of CDRISC was (59.50± 12.77).PSQI was negatively correlated with FFMQ and CD-RISC (r=-0.29,-0.24;both P＜0.01),and there was a significant positive correlation between FFMQ and CD-RISC (r=0.48,P＜0.01).The factors of FFMQ associated with CD-RISC were followed by the description of mindfulness,the action of awareness,the non-reaction and the observation of mindfulness.The multi-linear regression analysis showed that resilience played a part mediating role between mindfulness and sleep quality,with a mediating effect of 20.9％.Conclusion Mindfulness has a positive impact on the quality of sleep of medical staff through resilience.

Objective:To investigate the efficacy of Kechuanning combined with western medicine on acute exacerbation of chronic obstructive pulmonary disease (AECOPD) and its effects on serum amyloid A, interleukin 1β and procalcitonin levels. Methods:A total of 104 patients with AECOPD who received treatment in Yongkang Hospital of Traditional Chinese Medicine from January 2019 to December 2020 were included in this study. They were randomly assigned to receive either symptomatic treatment with western medicine alone ( n = 52, control group) or symptomatic treatment with western medicine combined with Kechuanning ( n = 52, observation group). Therapeutic effects, latency to clinical symptom relief, pre- and post-treatment pulmonary function, serum inflammatory factor levels, and blood gas analysis indexes were compared between the two groups. Results:Total response rate in the observation group was significantly higher than that in the control group [86.54% (45/52) vs. 67.31%(35/52), χ2 = 4.99, P < 0.05]. Latency to rale disappearance, latency to cough disappearance, length of hospital stay in the observation group were (8.25 ± 1.38) days, (10.05 ± 1.53) days, and (12.65 ± 2.28) days, which were significantly shorter than those in the control group [(9.41 ± 1.46) days, (12.19 ± 1.61) days, (14.36 ± 2.14) days, t = 4.16, 6.98, 3.61, all P < 0.05]. After treatment, forced vital capacity (FVC), forced expiratory volume in the first second (FEV 1), and FEV 1/FVC value in the observation group were (1.88 ± 0.5) L, (64.13 ± 5.72)%pred, (59.43 ± 5.57)%, respectively, which were significantly higher than those in the control group [(1.65 ± 0.51) L, (60.22 ± 5.60)% pred, (54.16 ± 5.19)%, t = 2.17, 3.52, 4.99, all P < 0.05]. Arterial partial pressure of oxygen (PaO 2) and blood oxygen saturation (SpO 2) in the observation group were (9.18 ± 0.89) kPa and (96.26 ± 2.13)%, respectively, which were significantly higher than those in the control group [(8.74 ± 0.76) kPa, (94.07 ± 2.08)%, t = 2.71, 5.305, both P < 0.05]. Partial pressure of carbon dioxide (PaCO 2) in the observation group was significantly lower than that in the control group [(7.32 ± 0.27) kPa vs. (7.63 ± 0.32) kPa, t = 5.34, P < 0.05]. Serum amyloid protein, interleukin-1β and procalcitonin levels in the observation group were (43.84 ± 6.15) mg/L, (3.24 ± 0.51) μg/L, (1.55 ± 0.37) ng/L, respectively, which were significantly lower than those in the control group [(55.26 ± 3.46) mg/L, (4.19 ± 0.56) μg/L, (2.03 ± 0.46) ng/L, t = 9.23, 9.04, 5.86, all P < 0.05]. Conclusion:Kechuanning as an adjuvant therapy for AECOPD can greatly improve lung function and hypoxia, alleviate clinical symptoms, reduce inflammatory reactions, and have a definite clinical effect. The study is innovative and scientific and is worthy of clinical reference.

Objective:To preliminarily explore the long-term improvement of low-frequency deep brain stimulation (DBS) on the nucleus basalis of Meynert (NBM) in cognitive disorders, neuropsychiatric symptoms and sleep disorders of patients with early-onset severe Alzheimer's disease (AD).Methods:A retrospective study was performed; 18 patients with early-onset severe AD admitted to Department of Neurosurgery, First Medical Center of PLA General Hospital from January 2016 to December 2022 were included. These patients were divided into NBM-DBS group and control group according to different treatments; 6 patients received low-frequency NBM-DBS on basis of conservative treatments; 12 patients accepted conservative treatments. Changes in Brief Mental State Examination (MMSE), Montreal Cognitive Assessment (MoCA), Neuropsychiatric Inventory (NPI), Hamilton Depression Rating Scale (HAMD), Becker-Lavanson Mania Scale (BRMS), Pittsburgh Sleep Quality Index (PSQI), and Zarit Caregiver Burden Interview (ZBI) were observed before treatment and 1 year after follow up.Results:MMSE and MoCA scores 1 year after follow up obviously reduced compared with those before treatment in both NBM-DBS and control patients; MMSE and MoCA scores in NBM-DBS patients showed no significant differences between 1 year after follow up and before treatment ( P>0.05), while significant differences were noted in the control group between 1 year after follow-up and before treatment ( P<0.05); and no significant differences in MMSE and MoCA scores were noted between the 2 groups 1 year after follow up ( P>0.05). NPI, HAMD, BRMS and ZBI scores in the NBM-DBS group 1 year after follow up were significantly different compared with those before treatment ( P<0.05); no significant differences were noted in NPI, HAMD and ZBI scores in the control group between 1 year after follow up and before treatment ( P>0.05), while significant difference was noted in BRMS scores ( P<0.05); significant differences in NPI, HAMD, BRMS and ZBI scores were noted between the 2 groups 1 year after follow up ( P<0.05). Conclusion:Low-frequency NBM-DBS is not only effective in improving cognitive disorders, but also effective in improving neuropsychiatric symptoms and sleep disorders, as well as reducing caregiver burden in patients with early-onset severe AD.