Objective To study the induction and detection methods of Staphylococcus aureus viable but non-culturable state (VBNC).Methods Staphylococcus aureus (ATCC13565) of 107 cfu/ml was induced by low temperature at 4 ℃,freezing at-20 ℃ and adding copper ions from 0.01 mmol/L to 0.05 mmol/L,Staphylococcus aureus was tested,and explored the resuscitation conditions by stepwise heating-heating and chemokinesis.Results Frozen 72 h at the temperature of-20 ℃ or added 0.015 mmol/L Cu2+ for 4 days of culture could induce staphylococcus aureus into the state of VBNC.Adding 0.5% tween-20 or 1% catalase for 24 h of culture could make the strain in VBNC achieve recovery.Conclusion Staphylococcus aureus can be induced into the VBNC and the recovered Staphylococcus aureus is the same as the normal bacteria in colony morphology and physiological and biochemical response in the common test medium.

AIM　To study the effect of desipramin e (DMI) on proliferation inhibition and apoptosis induction of rat glioma C6 cel ls. METHODS　Cell proliferation w as measured by MTT colorimetric assay and cells undergoing apoptosis were determ ined by electron microscope and flow cytometry. The expression of bcl-2 was eva luated by immunohistochemistry. RESULTS　DMI could result in the c oncentration-dependent inhibition of C6 cell proliferation and lead to arrest i n G0～G1 phase of cell cycle. The value of IC50 and 95% confidence lim its were 20.7(17.3～24.2) μmol*L－1.DMI（40 μmol*L－1）-indu ced apoptosis showed classical apoptotic morphology and the hypodiploid peak ap peared on the histogram of FCM in a concentration-dependent manner, which could be abrogated by cycloheximide(1.8 μmol*L－1). Meanwhile, DMI (10 μmol *L－1) could down-regulate the expression of apoptosis associated gene b cl-2. CONCLUSION　DMI could inhibit cell proliferation in a con centration dependent manner and induce typical apoptosis of C6 cells.

AIM To study the effect of desipramine (DMI ) on proliferation inhibition and apoptosis induction of rat glioma C6 cells. METH ODS Cell proliferation was measured by MTT col- orimetric assay and cells undergoing apoptosis were determined by electron microscope and flow cytometry. The expression of hcf-2 was evaluated by immunohistochemistry. RESULTS DMI could result in the concentration- dependent inhibition of C6 cell proliferation and lead to arrest in GO - G1 phase of cell cycle. The value of Ica and 95% confidence limits were 20.7(17 .3~24 .2) ?mol?L~ 1. DMI(40 ?mol? L-l )-induced apoptosis showed classical apoptotic morphology and the hypodiploid peak appeared on the histogram of FCM in a concentration- dependent man ner, which could be abrogated by cycloheximide(1. 8 ?mol? L- １ ). Meanwhile, DMI (10 ?mol? L- 1 ) could down-regulate the expression of apoptosis associated gene hcl-2. CONCLUSION DMI could inhibit cell proliferation in a concentration dependent manner and induce typical apoptosis of C6 cells.

Cultivation of academic type of medical graduate students is an important part of medical education system in our country and the individualized cultivation is an important way of training.Frist,the individualized cultivation of academic type of medical graduate students requires that supervisors fully understand the characteristics of the graduate students based on supervisors' subjective feelings and the result of scientific research test of the students.and then divide the students into three types(type A,B and C).With reference to the PDCA cycle management model,the supervisors carry out the individualized cultivation for different types of students in various links of the PDCA cycle,in order to encourage excellent students,urge the average students,and help the students who are behind.From the result of the individualized cultivation of academic type of medical graduate students in single research team,the proportion of students type rose from 2.9％ (type A),65.7％ (type B) and 31.4％(type C) at the beginning to 23.5％(type A),68.6％(type B) and 7.9％(type C) at the time of graduation.The research output is huge,so as to promote the quality of cultivation of academic type of medical graduate students.

OBJECTIVETo observe the effect of 3'-methoxy puerarin on cerebral infarction volume and free radical change of focal cerebral ischemia-reperfusion injury in rats and discuss the protective effect of 3'-methoxy puerarin on the cerebral ischemic/ reperfusion injury.

METHODThe thread method was used to induce middle cerebral artery embolization, to establish the focal cerebral ischemia-reperfusion model. Rats were divided into five groups: the sham and model group, the two-dose group (5, 10 mg x kg(-1) x d(-1)) of 3'-methoxy puerarin and nimodipine group (5 mg x kg(-1) x d(-1)). The behavior changes and volume of cerebral infarction were observed, and the leves of SOD and the content of MDA were measured.

RESULT3'-methoxy puerarin could significantly improve the symptoms of neurological deficit and reduce the infarct volume, and increased SOD activity and reduced the content of MDA of cortex in cerebral ischemia-reperfusion injury rat, the action of 10 mg x kg(-1) of 3'-methoxy puerarin is more remarkable.

CONCLUSION3'-methoxy puerarin has protective effect on focal cerebral ischemia-reperfusion injury in rat.

Animals ; Brain Ischemia ; drug therapy ; prevention & control ; Disease Models, Animal ; Female ; Humans ; Isoflavones ; pharmacology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; prevention & control

Objective To optimize the simultaneous extracting technique of saponins and total flavonoids from licorice (gancao).Methods Ammonia concentration (A), ethanol concentration (B), reflux time(C), and liquid/solid ratio(D) were set as the independent variables in this single factor experiment.Four factors and five levels of central composite design (CCD) in response surface methods were used to determine the content of saponins and total flavonoids in licorice.This study used ultraviolet spectrophotometric method to measure saponins and total flavonoids in licorice at the wave length of 252 nm and 510 nm respectively.The entropy weight method in the R language application was used to assign weight to the above two parameters.The three-layered model of BP neural network was established to test the effect of the number of hidden neurons (size).Finally, genetic algorithm was established to optimize the extraction techniques with real-coded program of R language.Results This method achieved the objective of testing requirements.There was a good linear relationship between saponins at 0.008~0.056 g/L, total flavonoids at 0.024~0.08 g/L, and light absorbance.This method set the neural network model with five hidden layer neurons.After optimizing the parameters of genetic algorithm, the extraction process of saponins and total flavonoids from licorice was optimized.The final optimal parameters were 0.62% ammonia, 64% ethanol, 1.8 h reflux time, and 12∶1 of liquid-solid ratio.In this optimal extraction condition, predictive value of this model was 191.65, and experimental average value was 188.90.The relative error was 1.43%, which demonstrated a good predictability of the neural network model and genetic algorithm.Conclusion This mathematical model to optimize the extraction techniques of saponins and total flavonoids from licorice is scientific and feasible.It also provids an innovative reference and approach to the multi-objective extraction techniques for identifying chemical compound and active ingredients of traditional Chinese medicine.

Objective:To investigate the regulatory mechanism of long non-coding RNA (lncRNA) NEF on T cell immune function in postmenopausal osteoporosis (PMOP) mice.Methods:Female Balb/c mice were used to construct OVX model ( n=46) and sham control group ( n=16) . Bone marrow mesenchymal stem cells (BM-SCs) from these two groups of mice were cultured. NEF recombinant expression vector (pIRSE2-NEF) was constructed and transfected into BMSCs. RT-qPCR was used to detect NEF and miR-21 levels in BMSCs cells in sham group, OVX group, and pIRSE2-NEF group. Luciferase gene report experiment was used to verify the binding effect of NEF and miR-21. The remaining 40 OVX mice were divided into 4 groups, including OVX group ( n=10) , pIRSE2-NEF injection group (pIRSE2-NEF group, n=10) , pIRSE2-NEF combined with PD-1 inhibitor group (pIRSE2-NEF+ PD-L1-IN-1 group, n=10) , and pIRSE2-NEF combined with miR-21 mimic (mimic) group (pIRSE2-NEF+ mimic group, n=10) . The remaining 10 mice in sham group were used as the control group. ELI-SA was used to detect the levels of IFN-γ, IL-2, IL-4, IL-13 and PD-1/PD-1L in peripheral blood. Flow cytometry was used to detect the shift of serum Treg-Th17 cell subsets. Results:Compared with the Sham group (1.01±0.04, 1.00±0.03) , the expression of NEF in BMSCs of OVX group was down-regulated (0.23±0.01) , and miR-21 was up-regulated (2.96±0.05) ( P<0.05) . Compared with OVX group (1.23±0.15, 5.20±0.31) , NEF in BMSCs cells of Pirse2-nef group was significantly up-regulated (6.83±0.35) ( P<0.05) , while miR-21 was down-regulated (0.29±0.11) ( P<0.05) .NEF has a direct binding base site with miR-21.The levels of IFN-γ (3.25±0.21) , IL-2 (2.44±0.06) and Th17/Treg ratio (3.18±0.65) in peripheral blood of mice in OVX group were significantly higher than those in Sham group (1.03±0.02, 1.00±0.01, 0.86±0.09) (all P<0.05) . The levels of IL-4 (0.45±0.02) , IL-13 (0.43±0.07) , PD-1 (0.24±0.03) and PD-1L (0.51±0.06) were significantly lower than those of Sham group (1.00±0.04, 1.00±0.02, 1.00±0.03, 1.00±0.00) ( P<0.05) ; Compared with OVX, IFN-γ (2.02±0.06) , IL-2 (0.88±0.01) and Th17/Treg ratio (1.43±0.22) in Pirse2-nef group were decreased. The levels of IL-4 (0.87±0.03) , IL-13 (0.84±0.07) , PD-1 (0.79±0.06) and PD-1L (0.77±0.06) were increased (all P<0.05) ; Compared with Pirse2-nef group, IFN-γ (2.89±0.06) , IL-2 (2.07±0.07) and Th17/Treg ratio (2.39±0.38) were increased in Pirse2-nef+ PD-L1-in-1 group. The levels of IL-4 (0.68±0.03) , IL-13 (0.76±0.08) , PD-1 (0.52±0.02) and PD-1L (0.83±0.04) were decreased (all P<0.05) . Moreover, the pIRSE2-NEF+ mimic group had the same adjustment effect as the pIRSE2-NEF+ PD-L1-IN-1 group. Conclusion:lncRNA-NEF improves immune imbalance and PD-1/PD-1L-mediated Treg-Th17 cell balance in postmenopausal osteoporosis mice by sponging miR-21.

Bullous pemphigoid (BP) can be comorbid with a variety of immune diseases, such as immune skin diseases (psoriasis, vitiligo, alopecia areata and various other immune bullous diseases) , immune digestive diseases (inflammatory bowel disease, primary biliary cirrhosis) , autoimmune thyroid diseases, autoimmune rheumatic diseases (rheumatoid arthritis, dermatomyositis, scleroderma and systemic lupus erythematosus) , immune renal diseases (immune nephropathy, renal allograft rejection) and acquired hemophilia A. The above comorbidities markedly affect the quality of life of and treatment options for patients. This review elaborates on currently reported immune diseases associated with BP and their concomitant mechanisms.

Objective:To investigate the epidemiological characteristics of COVID-19 caused by 2019-nCoV Delta variant in Shangyu district, Shaoxing of Zhejiang province in 2021, and provide evidence for the improvement of COVID-19 control and prevention measures.Methods:The incidence data of COVID-19 in Shangyu from December 7 to 21, 2021 was obtained from Shangyu District Center for Disease Control and Prevention. The epidemiological characteristics of the cases, i.e. the population, time and space distributions, were analyzed, and the incubation period and time-varying reproduction numbers ( R t) were calculated. Results:From December 7 to 21, 2021, a total of 380 COVID-19 cases caused by 2019-nCoV Delta variant were detected in Shangyu, the median age of the cases was 52 years, M ( Q1，Q3: 38, 61). The male to female ratio of the cases was 1∶1.25, and the cases were mainly workers (36.58%) and farmers (27.63%). The epidemic affected 9 townships (or community) of Shangyu, especially Caoe and Baiguan communities with the cases accounting for 57.10% and 31.53% respectively. The median incubation period of cases was 4.00 days, M ( Q1，Q3: 3.00, 5.75). The basic reproduction number ( R0) was 4.06, and the R t was 5.62 in early phase of the outbreak (the highest) and continuously decreased to less than 1.00 within 10 days after the detection of the outbreak. The number of COVID-19 cases decreased to 0 within 14 days after the outbreak (December 7-21), and the main detection methods were screening in centralized isolation (55.53%) and home isolation (40.00%). The infection rates of close contacts and secondary close contacts were 2.06% and 0.46% respectively. Conclusion:The epidemic of COVID-19 in Shangyu characterized by short incubation period, large number of infected people, and case clustering, suggesting the strong transmission of Delta variant (B.1.617.2). Comprehensive prevention and control measures, such as management of close contacts and secondary close contacts, and high-risk area, are essential for the rapid control of the epidemic.

In vitro diagnostic technology is an important aid for disease prevention,treatment,and prognostic monitoring.The de-velopment of its new technologies,methods and products is of great significance for improving medical quality and protecting public health.On the basis of exploring the mechanism of health impact of in vitro diagnostic technology,it takes dry chemical method and wet chemical method as an example,and compares the economy of the two diagnostic technologies for hyperkalemia diagnosis from the perspective of health system,with a view to draw references for the practical application and evaluation of new in vitro diagnostic techniques,methods and products in the future.