OBJECTIVETo explore the intergrating of N-isopropyl oxamate and serum protein and establish a high performance liquid chromatography (HPLC) detection method of N-isopropyl oxamate (specific inhibitor of testis-specific lactate dehydrogenase (LDH-C4)) in the blood of plateau pikas.

METHODSTwenty highland pika 150-200 g, were randomly divided into two groups (n = 10): control group and inhibitor group. Different concentrations of N-isopropyl oxamate were added to examine the intergrating of N-isopropyl oxamate and serum protein. In order to determine its concentration in the pika blood accurately, we used the method of adding trypsin to incubate the serum first, followed by trichloroacetic acid treatment and detecting by HPLC. Results: When the concentrations of N-isopropyl oxamate in the pika serum were added to 0.05 mmol/L, 0.1 mmol/L, 1 mmol/L, 10 mmol/L, 16.7 mmol/L, 33.3 mmol/L and 100 mmol/L, the intergrating rates between N-isopropyl oxamate and plateau pika serum were 100%, 100%, 100%, 86.84%, 54.11%, 40.10% and 20.18%, respectively. The method established in this paper was good on recovery rates, precision and stability. A good linearity was obtained in the range of 0.0125-0.25 mmol/L. When the concentrations of N-isopropyl oxamate in the serum were added to 0. 15 mmol/L,0.3 mmol/L and 1 mmol/L, the recovery rates were 98.05%, 98.98% and 98.12%, respectively; the precision relative standard deviation( RSD) of concentrations were 1.17%, 0.92% and 0.83%, respectively; the stability relative standard deviation (RSD) of concentrations were 1.38%, 1.40% and 0.88%, respectively. The repeatability RSD of the method was 1.76%. Quantitative limit was 0.0125 mmol/L.

CONCLUSIONN-isopropyl oxamate has a strong affinity with plateau pika serum protein that can't be accurately determined with common HIPLC method. It can be accurately determined in the blood by adding trypsinto digest the serum protein first, followed by adding trichloroacetic acid to precipitate the protein.

Animals ; Chromatography, High Pressure Liquid ; methods ; Lagomorpha ; Male ; Oxamic Acid ; analogs & derivatives ; blood

BACKGROUND:For patients in intensive care unit (ICU), mechanical ventilation is an effective treatment to survive from acute illness and improve survival rates. However, long periods of bed rest and restricted physical activity can result in side effects. This study aimed to investigate the feasibility of early rehabilitation therapy in patients with mechanical ventilation. METHODS:A randomized controlled trial was carried out. Sixty patients, with tracheal intubation or tracheostomy more than 48 hours and less than 72 hours, were admitted to the ICU of the Affiliated Hospital of Medical College, Qingdao University, from May 2010 to May 2012. These patients were randomly divided into a rehabilitation group and a control group. In the rehabilitation group, rehabilitation therapy was performed twice daily, and the training time and intensity were adjusted according to the condition of the patients. Early rehabilitation therapy included heading up actively, transferring from the supine position to sitting position, sitting at the edge of the bed, sitting in chair, transferring from sitting to standing, and ambulating bedside. The patient's body mass index, days to first out of bed, duration of mechanical ventilation, length of ICU stay, APACHE Ⅱ score, highest FiO2, lowest PaO2/FiO2 and hospital mortality of patients were all compared between the rehabilitation group and the control group. The differences between the two groups were compared using Student's t test. RESULTS:There was no significant difference in body mass index, APACHE Ⅱ score, highest FiO2, lowest PaO2/FiO2 and hospital mortality between the rehabilitation group and the control group (P>0.05). Patients in the rehabilitation group had shorter days to first out of bed (3.8±1.2 d vs. 7.3±2.8 d; P=0.00), duration of mechanical ventilation (5.6±2.1 d vs. 12.7±4.1 d; P=0.005) and length of ICU stay (12.7±4.1 d vs. 15.2±4.5 d; P=0.01) compared with the control group. CONCLUSION:Early rehabilitation therapy was feasible and effective in improving the outcomes of patients with mechanical ventilation.

ObjectiveTo explore the prevalence and spectrum of β-thalassemia mutations in Fujian province,and to provide a reference for prenatal diagnosis and genetic counseling in this population.Methods Two thousand three hundred and one blood samples were randomly selected from 9 different areas of Fujian province from May 2008 to December 2010.PCR and reverse dot blot hybridization (RDB) were adopted for detection of the 17 common types of mutation,and the frequency of each genotype of β-thalassemia mutations was calculated.The β-globin gene of unknown positive samples were analyzed directly with DNA sequencing.Results Three hundred and fifty-nine cases were detected with β-thalassemia mutations of the 2301 copy blood samples submitted,and the detection rate was 15.60％ (359/2301).Of the mutated genes,12 different mutations were identified,namely IVS-2-654(C→T),CD41-42(-TCTT),CD17(A→T),-28(A→G),CD27-28(+C),CD26(G→A),CD71-72(+A),IVS-1-1(G→T),CD43(G→T),-29(A→G),initiation codon ATG→AGG and CD36(-C).Mutation frequencies were 46.54％ (175/376),33.24％ (125/376),9.31％ (35/376),5.05％ (19/376),2.13％(8/376),1.33％(5/376),0.80％(3/376),0.27％(1/376),0.27％(1/376),0.27％(1/376),0.53％(2/376),and 0.27％(1/376),respectively.The most common mutations were IVS-2-654 (C→T) and CD41-42 (-TCTT),which accounted for 79.78％(300/376) of total genetic mutations.In addition,a novel β-globin gene mutation CD36 (-C) allele was detected for the first time,the deletion of a nucleotide C at code 36 within exon 2 lead to a frameshift mutation that could result in a premature termination at code 60.Conclusions β-thalassemia mutations in Fujian province are complex with significant genetic heterogeneity.We present for the first time the detection of a new β-thalassemia mutation in the population:CD36(-C),which provides valuable information for genetic counseling and prenatal diagnosis in Fujian province.

OBJECTIVETo explore the feasibility of IGF2 imprinting system in target gene therapy for tumors.

METHODSThe mouse H19 enhancer, DMD and promoter H19 were amplified by PCR from mouse genomic DNA and then cloned into the plasmid pDC312. The EGFP and DT-A fragments were amplified by PCR and cloned into the recombinant plasmid, and then the shuttle plasmid were transfected into HEK293 cells together with the adenoviral vector Ad5, namely, Ad-EGFP and Ad-DT-A. Adenovirus hexon gene expression was applied to confirm the presence of adenovirus infections. The effect of the IGF2 imprinting system was tested by fluorescence microscopy. RT-PCR and Western blotting after transfection of the recombinant adenoviral vectors into cancer cells were used to show loss of IGF2 imprinting (LOI) and maintenance of IGF2 imprinting (MOI), respectively. The anti-tumor effect was assessed by MTT and flow cytometry after the HCT-8 (LOI). Human breast cancer cell line MCF-7 (MOI) and human normal gastric epithelial GES-1 (MOI) cell line were transfected with Ad-DT-A in vitro. The anti-tumor effect was detected by injecting the Ad-DT-A in nude mice carrying HCT-8 tumors.

RESULTSThe expression of EGFP protein, DT-A mRNA and DT-A protein were seen to be positive only in the HCT-8 tumor cell line. Infection with Ad-DT-A resulted in obviously growth inhibition in HCT-8 cells (75.4 ± 6.4)% compared with that in the control group, and increased the percentage of apoptosis in the HCT-8 cells (20.8 ± 5.9)%. The anti-tumor effect was further confirmed by injecting the recombinant adenoviruses in HCT-8 tumor-bearing nude mice, and the results showed that the Ad-DT-A inhibited the tumor growth, with on inhibition rate of 36.4%.

CONCLUSIONSThe recombinant adenoviruses carrying IGF2 imprinting system and DT-A gene have been successfully constructed, while Ad-DT-A can effectively kill the tumor cells showing loss of IGF2 imprinting. It might play an important role in future target gene therapy against malignant tumors based on loss of IGF2 imprinting.

Adenoviridae ; genetics ; Animals ; Apoptosis ; Breast Neoplasms ; genetics ; pathology ; Colonic Neoplasms ; genetics ; pathology ; therapy ; Diphtheria Toxin ; biosynthesis ; genetics ; Female ; Genetic Therapy ; methods ; Genetic Vectors ; Genomic Imprinting ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Insulin-Like Growth Factor II ; genetics ; metabolism ; MCF-7 Cells ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Peptide Fragments ; biosynthesis ; genetics ; Plasmids ; RNA, Messenger ; metabolism ; Random Allocation ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

Objective To find out the arsenic pollution in Yangzonghai lake and its influence to the surrounding drinking water source and the local food and to provide a scientific basis for arsenic pollution control,drinking water and food safety. Methods Arsenic monitoring were carried out in 10 points of Yangzonghai lake,45 water sources within 5 km of Yangzonghai lake, and locally produced food. Results From September 2008 to December 2009, the water arsenic concentrations of Yangzonghai lake ranged 0.06 - 0.18 mg/L, the average arsenic concentrations were 0.12 mg/L and 0.10 mg/L for 2008 and 2009, respectively, the value of 2009 was higher, and the difference was statistically significant(t = 3.284, P ＜ 0.05). There were no significant difference in the average of arsenic concentration of Yangzonghai lake between the dry and rainy season in 2009(t = 0.905, P ＞0.05). The arsenic concentrations from the No. 3 spring water ranged from 0.12 - 30.36 mg/L. After a peak value early in October 2008, the No. 3 spring water showed a downward trend month by month. In 2008 the average arsenic concentration was 23.92 mg/L, 2009 was 2.41 mg/L, down 89.92% over the previous year, the difference was statistically significant(t = 9.582, P ＜ 0.05 ). In January and February 2009, the average arsenic concentration (11.96 mg/L) was significantly higher than those of March-December(0.50 mg/L, t = 57.759, P＜ 0.05). In Yangzong town, the arsenic concentration in the drinking water and river met the national drinking water health standards. Among 78 samples monitored of the locally produced food in Yangzong town, 77 passed, with a pass rate of 98.72%. Conclusions Arsenic contamination has been effectively controlled. The drinking water and local food production has not been contaminated by arsenic except aquatic products in Yangzonghai Lake. It is recommended to take effective measures to prevent water contamination.

OBJECTIVETo investigate the sleep quality and related factors among medical students in China, understand the association between dormitory environment and sleep quality, and provide evidence and recommendations for sleep hygiene intervention.

METHODSA total of 555 undergraduate students were selected from a medical school of an university in Beijing through stratified-cluster random-sampling to conduct a questionnaire survey by using Chinese version of Pittsburgh Sleep Quality Index (PSQI) and self-designed questionnaire. Analyses were performed by using multiple logistic regression model as well as multilevel linear regression model.

RESULTSThe prevalence of sleep disorder was 29.1%(149/512), and 39.1%(200/512) of the students reported that the sleep quality was influenced by dormitory environment. PSQI score was negatively correlated with self-reported rating of dormitory environment (γs=-0.310, P<0.001). Logistic regression analysis showed the related factors of sleep disorder included grade, sleep regularity, self-rated health status, pressures of school work and employment, as well as dormitory environment. RESULTS of multilevel regression analysis also indicated that perception on dormitory environment (individual level) was associated with sleep quality with the dormitory level random effects under control (b=-0.619, P<0.001).

CONCLUSIONSThe prevalence of sleep disorder was high in medical students, which was associated with multiple factors. Dormitory environment should be taken into consideration when the interventions are taken to improve the sleep quality of students.

Beijing ; epidemiology ; Housing ; Humans ; Logistic Models ; Prevalence ; Risk Factors ; Schools, Medical ; Self Report ; Sleep Wake Disorders ; epidemiology ; Social Environment ; Students, Medical ; psychology ; statistics & numerical data

To clone mouse phage antibodies against H-Y antigen from a phage antibody library, three cycles of affinity enrichment of the mouse phage antibody library with male spleen cells and two cycles of nonspecific absorption with female spleen cells were performed. The presence of mouse Fab on the phage surface was determined by ELISA and sequence analysis. 9 of 15 strains can bind to male spleen cells with the specific activity. Recombination rate of the phage antibody library clones is 60%. Sequence analysis of the PCR products of plasmid DNA of E5 clones show VH and Vkappa had common characteristics shared by other known variable region of antibodies. The mouse phage Fab antibody could be used for identifying H-Y antigen, and for the development of sex determination of early embryos in mammals.
Animals ; Base Sequence ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Female ; H-Y Antigen ; analysis ; Immunoglobulin Fab Fragments ; genetics ; immunology ; Isoantibodies ; genetics ; immunology ; Male ; Mice ; Molecular Sequence Data ; Peptide Library

To study the constituents from the chloroform extract of the roots of Lasianthus acuminatissimus Merr., various chromatographic techniques were used to separate and purify the constituents. The structure was established on the basis of ID, 2D NMR and HRMS spectroscopic analysis. A new compound was isolated and identified, which was 3, 8-dihydroxy-1-methoxy-2-methoxymethyl-9,10-anthraquinone (I). Compound I is a new anthraquinone, namely lasianthurin.
Anthraquinones ; chemistry ; isolation & purification ; Chromatography, Thin Layer ; Magnetic Resonance Spectroscopy ; Molecular Conformation ; Molecular Structure ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rubiaceae ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Objective:To understand the epidemiological and clinical characteristics of patients with tsutsugamushi disease in Zaozhuang City, Shandong Province, and to provide a basis for formulating effective diagnosis and treatment plan of tsutsugamushi disease. Methods:Retrospective analysis was carried out to collect clinical data of 38 patients with tsutsugamushi disease admitted to Zaozhuang Municipal Hospital of Shandong Province from 2018 to 2020. The epidemiological characteristics, clinical manifestations, laboratory examination results and drug treatment of patients with tsutsugamushi disease were analyzed. Results:Among the 38 patients with tsutsugamushi disease, 15 were males and 23 were females, aged (58.58 ± 14.47) years old; the occupation distribution was mainly farmers, accounting for 92.11% (35/38); the onset time was concentrated in October and November, accounting for 100.00% (38/38); the regional distribution was mainly in Yicheng District, accounting for 34.21% (13/38); 38 patients had a history of grassland/forest contact. All patients had fever, and other main symptoms were eschar (97.37%, 37/38), headache (68.42%, 26/38), fatigue (42.11%, 16/38), and whole body aches (36.84%, 14/38), etc. The results of laboratory examination mainly showed that C-reactive protein increased (89.47%, 34/38), procalcitonin increased (81.58%, 31/38), abnormal liver function (78.95%, 30/38), D-dimer increased (71.05%, 27/38), eosinophils decreased (60.53%, 23/38), and erythrocyte sedimentation rate increased (57.89%, 22/38). All 38 patients were cured after treated with doxycycline or azithromycin. Conclusions:The high incidence time of tsutsugamushi disease in Zaozhuang City is autumn. The clinical symptoms are mainly fever, eschar and headache. Doxycycline and azithromycin are the first choices for treatment of the disease.

Since the outbreak of novel coronavirus pneumonia, the World Health Organization and the National Health Commission have issued multiple guidance documents.Medical facilities across the nation have actively implemented the countermeasures in response to the epidemic.However, in the real scenario of prevention and control, hospital infection remains a great concern.It is crucial to formulate measures and procedures based on standard prevention and transmission route prevention to avoid cross infection between medical workers and patients.Based on the practice, we summarize the institutional management, layout process, prevention and control of hospital infection in the emergency wards for further discussion and improvement.