This paper analyzes the factors that affect students' autonomous learning from the aspects of teaching interaction,the binding force of curriculum to students,the application of network and electronic products.On the basis of this,we put forward that taking the molecular biology course of biotechnology major and bio-pharmacy Major as a teaching reform template in our university,and based on the network teaching platform of molecular biology,adopt flipped classroom as the main form to improve students' autonomous learning ability,and promote their ability of lifelong learning,so that they can adapt to the development of pharmaceutical biotechnology industry.

In line with the new WHO definition for health and reference of comprehensive health assessments of elderly overseas, the paper screened and set the weight for comprehensive health assessment indicators for military officers, establishing a general health assessment model as a result 402 military officers were subject to Comprehensive Health Assessments, and the outcomes were compared with subjective assessment of experts. The results of both methods are highly coincidental. It proves Comprehensive Health Assessment as a scientific quantitative health assessment method, with promising perspectives in health management in the future.

Objective To examine the relationship between smoking and tobacco-related mortality in a cohort of the aged Chinese armymen.Methods A cohort study was carried out in Xi'an,China from 1987 to 2005.A total of 1 268 retired male military cadres aged 55 or older were examined in 1987 and followed up in the next 18 years.At baseline,388 men were non-smokers,461 were former smokers,and 419 were current smokers.The current smoking rate was 33.04%,Quitted rate was 52.39%.The terminal main observation items included all-cause and tobacco-associated mortality.Results Up to Jun 30,2005,a total of 491 subjects died.Compared with non-smokers,the relative risks(95% confidence intervals(CI)) of ever-smokers for all-cause deaths,chronic obstructive pulmonary disease(COPD),lung cancer,and coronary heart disease(CHD) were 1.24(1.01-1.53),1.91(1.06-3.46),2.91(1.36-6.23),and 1.15(0.68-1.93),respectively.Compared with current smokers,the risks of total mortality and death from lung cancer for former smokers were decreased by 66.7% and 14.98%,respectively.Conclusions Cigarette smoking was a major death risk factor in the cohort and quitting smoking could save lives.

Objective:To investigate the perfection and improvement of the execution of integrative medicine therapy in severe tetanus therapy, to successfully control tetanus severe spasms, autonomic dysfunction and prevent lethal side-effect of prolong and high-dosage sedative-muscle-relaxant therapy, resulted in significant reduction of mortality of tetanus.Methods:Symptoms, treatments and outcome of tetanus patients admitted to Peking University Third Hospital from 1965 to 2020 were reviewed. Patients were classified with Ablett classification. The cases of Ablett grade Ⅲ and Ⅳ were severe tetanus. The patients were divided into two groups according to whether they were treated together with traditional Chinese medicine (TCM) simultaneously during the standard tetanus treatment; the patients in the TCM group were divided into the tetanus TCM medication group and the non tetanus TCM medication group according to the medicine provided whether was in accord with the conventional tetanus TCM prescriptions. The mortality of each group was calculated. In addition, one survived and one deceased case with severe convulsion, autonomic nerve dysfunction (Ablett grade Ⅳ) were selected, combined with the treatment methods and curative effects, the types, use methods and outcomes of Chinese and Western medicine were analyzed.Results:The 46 tetanus cases were treated with Western medicine. Twenty-two of them, TCM were applied. Fifteen of the 22 cases took the TCM prescription which was accord with the conventional tetanus prescription. The mortality of the 46 cases was 21.7% (10/46). The number of non-TCM group was 24 cases, with mortality of 20.8% (5/24); 1 case was Ablett Ⅱ, 1 was Ablett Ⅲ and 3 were Ablett Ⅳ. The number of the TCM group was 22 cases, with mortality of 22.7% (5/22), 2 cases were Ablett Ⅲ, 3 were Ablett Ⅳ. The TCM prescription of these 5 deceased cases was not directed towards tetanus. The tetanus TCM medication group was 15 cases, with no mortality. Case analyses: case 1 was intubated because of severe spasms. Autonomic dysfunction occurred on the 8th day after admission. Esmolol with increasing the dosage of the sedatives and muscle relaxant, was not effective. Tetanus TCM was applied after 2 days of autonomic dysfunction happened. Autonomic dysfunction was then under controlled on the 2nd day post-TCM. She was recovery and discharged after 4 weeks. Case 2, also was intubated because of severe spasms. Autonomic dysfunction happened on the 3rd day after admission, and failed to be controlled by large-dose sedatives, muscle relaxant, and Esmolol. After 8 days of persistent autonomic dysfunction, tetanus TCM was applied and autonomic dysfunction was under controlled on the 2nd day post-TCM administration. Large dosage of muscle-relaxant was applied continuously. After 5 days' administration of TCM, the TCM was withdrew. One day after the withdrawal of TCM, respiratory and cardiac arrest happened because of the diffused bronchiole obstruction with pulmonary secretions loading.Conclusion:Based on the precise and real-time diagnosis of the state of the disease, integrative medicine therapy with an overall analysis tetanus TCM prescription, is the key of declining tetanus mortality.

OBJECTIVEAccurate and reliable assessment of renal function is important in the management of children with chronic kidney disease (CKD). Glomerular filtration rate (GFR) is the best index of assessing kidney function. For assessment of GFR, both gold standard tests and prediction equations have been used. The well-known 24-hour endogenous creatinine clearance (Ccr), the Schwartz formula and the Filler formula are increasingly used in daily clinical practice. However, there are few studies on the applicability of these prediction equations for estimating GFR in Chinese children with CKD. The aim of this study was to compare these prediction equations estimating GFR with an isotope clearance method [isotope glomerular filtration rate (rGFR)] in such patients.

METHODChildren aged 1-16 years who underwent isotope (99m)Tc-diethylenetriaminepentaacetic acid ((99m)Tc-DTPA) GFR testing (Gates' method) between the year of 2002 and 2005 were studied retrospectively. GFR was estimated using: (1) 24-hour Ccr, which was calculated using the standard formula: [urine creatinine (milligrammes per millilitre) × 24-hour urine volume/serum creatinine (milligrammes per millilitre) × 1440] × [1.73 (m(2))/body surface area (m(2))]; (2) the Schwartz formula, which is: eGFR (ml/min per 1.73 m(2)) = k × height (centimetres)/serum creatinine (micromoles per litre), where k is 62 in males at 13 years of age and older, 40 in infants, and 49 in all other children; and (3) the Filler formula, which is: logGFR = 1.962 + [1.123 × log(1/Cys C)], where cystatin C is measured in milligrammes per litre. Serum and urinary creatinine levels were detected by alkaline kinetic method. Serum cystatin C was analysed by particle-enhanced immunoturbidimetric assay. Bias and precision were evaluated.

RESULTThirty subjects (18 males and 12 females; mean age 9.4 years) fulfilling both inclusion criteria and exclusion criteria were included in this study. The mean (SD) rGFR was 81.57 (36.92) ml/min per 1.73 m(2); 18 subjects were in CKD stage I, 8 in CKD stage II, 8 in CKD stage III, and 1 in CKD stage IV. Only the mean 24 h Ccr-eGFR was slightly higher than rGFR (0.4 ml/min per 1.73 m(2) higher). Within 95% limits of agreement, the maximum absolute value of bias was about 50 ml/min per 1.73 m(2). Accuracy (estimated GFR values within ± 30% of rGFR) for all formulae was poor, ranging from 23.3% to 43.3%. All formulae overestimate or underestimate rGFR in different CKD stages.

CONCLUSIONIn Chinese children with CKD, there was a significant difference between measured GFR and estimated GFR using 24h Ccr, Schwartz formula and Filler formula. More suitable GFR predictive equations to assess glomerular function of such patients should be developed.

Adolescent ; Child ; Child, Preschool ; Female ; Glomerular Filtration Rate ; Humans ; Infant ; Kidney Failure, Chronic ; physiopathology ; Kidney Function Tests ; Male ; Renal Insufficiency, Chronic ; physiopathology

The experiment was conducted by directed evolution strategy (error-prone PCR) to improve the activity of aflatoxin detoxifzyme with the high-throughput horse radish peroxidas and recessive brilliant green (HRP-RBG) screening system. We built up a mutant library to the order of 10(4). Two rounds of EP-PCR and HRP-RBG screening were used to obtain three optimum mutant strains A1773, A1476 and A2863. We found that mutant A1773 had upper temperature tolerance of 70 degrees C and that its enzyme activity was 6.5 times higher than that of the parent strain. Mutant strains A1476 worked well at pH 4.0 and its enzyme activity was 21 times higher than that of the parent strain. Mutant A2863 worked well at pH 4.0 and pH 7.5, and its enzyme activity was 12.6 times higher than that of the parent strain. With DNA sequencing we found that mutant A1773 revealed two amino acid substitutions, Glu127Lys and Gln613Arg. Mutant A1476 revealed four amino acid substitutions: Ser46Pro, Lys221Gln, Ile307Leu and Asn471lle. Mutant A2863 revealed four amino acid substitutions: Gly73Ser, Ile307Leu, Va1596Ala and Gln613Arg. The results provided a useful illustration for the deep understanding of the relationship between the function and structure of aflatoxin detoxifzyme.
Aflatoxin B1 ; antagonists & inhibitors ; chemistry ; Amino Acid Substitution ; Directed Molecular Evolution ; Enzyme Activation ; Enzyme Stability ; Multienzyme Complexes ; genetics ; metabolism ; Mutant Proteins ; genetics ; metabolism ; Point Mutation ; Polymerase Chain Reaction ; methods ; Protein Engineering

OBJECTIVE:To establish HPLC-quadrupole/electrostatic field orbitrap high resolution mass spectrometry for rapid determination of aconitine,mesaconitine,hypaconitine,benzoylaconitine,benzoylmesaconine and benzoylhypacoitine in rat plasma. METHODS:Internal standard lappaconitine was added into plasma sample,and methanol precipitated protein was used for pretreatment. HPLC-quadrupole/electrostatic field orbitrap high resolution mass spectrometry was adopted. HPLC condition was as follows as Sinochrom ODS-BP C18column,mobile phase consisted of acetonitrile-1% formic acid solution(50:50,V/V),the flow rate of 0.6 mL/min,sample size of 10 μL,column temperature of 25 ℃,automatic sampler temperature of 4 ℃. Mass spectrum scanning mode was full ion monitoring model,positive ion acquisition,mass charge ratios(m/z)of ion were 646.32(aconitine), 632.30(mesaconitine),616.31(hypaconitine),604.31(benzoylaconitine),590.29(benzoylmesaconine),574.30(benzoylhypacoitine), 585.31(internal standard). Six male Wistar rats were collected and given single dose of total alkaloid extract of Aconitum carmichaeli(4 mg/kg)intragastrically. Blood samples were collected before medication(0 h)and 0.5,0.75,1.25,1.5,2,4,6, 8,10,24 h after medication. Plasma concentration was determined and pharmacokinetic parameters were calculated by using PK-Solver V2.0 software. RESULTS:The linear range of 6 kinds of aconitum alkaloids in plasma were 0.1-10 μ g/L(r>0.992). The limit of quantitation was 0.1 μ g/L. Average recovery was higher than 75%,RSDs of intra-day and inter-day,matrix effects,stability test were all lower than 15%. The tmaxof 6 kinds of aconite alkaloids were about 1.2 h;t1/2were about 10 h;cmaxof monoestertype aconite alkaloids were higher than those of diester-type aconite alkaloids. CONCLUSIONS:Established HPLC-quadrupole/electrostatic field orbitrap high resolution mass spectrometry is accurate,sensitive,simple and rapid, and can be used for plasma concentration monitoring of 6 kinds of aconitum alkaloids.

Cerebral small vessel disease (CSVD) is a series of clinical syndrome characterized by the damage of small blood vessels in the brain caused by a variety of causes. Although the pathophysiological mechanisms of CSVD are not fully understood, increasing evidence suggests that matrix metalloproteinase-9 plays an important role in the occurrence and development of CSVD.

In recent years, the incidence of cerebral small vessel disease (CSVD) has been increasing with the aging of the population, and the cognitive impairment caused by it has brought huge burden to patients and their families. As a novel inflammatory biomarker, lipoprotein-associated phospholipase A 2 (Lp-PLA 2) directly participates in the pathogenesis of cognitive impairment in patients with CSVD by regulating circulatory vascular injury and neuroinflammation, and is expected to become a predictive indicator and therapeutic target for CSVD.

The molecular biology experiment course plays a bridge role in the biotechnology curriculum system, and has important effect in training students' experimental operation skills, analyzing difficulties and solving problems. This article takes the biotechnology major of local medical colleges as an example to analyze and elaborate the assessment method reform and effect in the teaching process of molecular biology experimental course, aiming at promoting the teaching level of molecular biology experimental courses and improving the training quality of biotechnology professionals.