Nowadays a large number of domestic researches have shown the degree of learning burnout,the causes,manifestations and the distribution,but there are only one or two cases focusing on empirical research of Group Counseling.In Eriksson's view,the university period is a critical stage to obtain a sense of intimacy and avoid isolation,and the group counseling have great significance in helping students improve the character and get a sense of intimacy.Therefore,based on the literature and interviews,the author use in this study the cognitive-behavioral group therapy to help the students form a new cognitive concepts,behavior and attitude,and improve their self-efficacy.

BACKGROUND:Conventional methods,including trypsin digestion and cells transfer using single call suspension,have many drawbacks,which limit the development of bone tissue engineering.OBJECTIVE:To culture bone marrow stromal stem calls,induce osteoblastic differentiation,and prepare cell sheets.METHODS:Canine bone marrow stromal calls were isolated by density gradient centrifugation technique,inoculated into DMEM medium,and induced to differentiate into osteoblasts.Complete call sheets were harvested by call sheet engineering based on the temperature change of temperature-responsive medium.RESULTS AND CONCLUSION:Immediately after inoculation,primary calls were scattered on the bottom of culture flask,presenting a transparent spherical body with a good refractive capacity.At 12 hours,calls exhibited a long shuttle shape,reached complete confluency,and grew in a whirlpool-like fashion.After osteoblastic induction,the majority of bone marrow stromal stem calls appeared tetragonal,polygonal,and squamose.At 21-28 days,round or oval-shaped calcified nodules formed.When the bone marrow stromal stem calls in the temperature-responsive culture dishes were cooled below the critical temperature 32℃,cells were gradually detached from the bottom of culture flask and formed complete bone marrow stromal stem call sheets.These findings indicate that density gradient cantrifugation technique can be used to successfully isolate and culture canine bone marrow stromal stem cells to differentiate into osteoblasts and call sheet engineering enables to harvest complete bone marrow stromal stem call sheets.

BACKGROUND:There are some disadvantages in harvesting and transferring cells in the traditional tissue engineering technique,and it is difficult to form dense tissues,which significantly limits the development of tissue engineering.OBJECTIVE:To explore the culture and fabrication of dog bone marrow mesenchymal stem cell(BMSC)sheet in vitro.METHODS:Bone marrow was extracted from dogs following anesthesia.BMSCs were isolated with the method of density gradient centrifugation in vitro.BMSCs at passage 4 at a density of 1×10~9/L were incubated in the temperature-responsive culture dishes with a diameter of 3.5 cm,and cultured in an incubator at 37 ℃,5% CO_2 and saturated humidity.The temperature of the incubator was changed from to 37 V to 20 ℃ to prepare BMSCs cell sheet for 20 minutes.Cell morphological changes and cell sheet formation were observed under an inverted microscope.RESULTS AND CONCLUSION:Dog BMSCs following 24 hours of primary culture presented ellipse or polygonal shape.Most cells adhered at hour 72,and cell colonies were visible at day 7.Cells showed long spindle and completely confluence at day 12,with unclear boundary.BMSCs in the temperature-responsive culture dishes presented short spindle shape,and gradually separated from the dish bottom,forming entire cell sheet containing extracellular matrix at 20 V.These verified that dog BMSCs can be effectively obtained with method of density gradient centrifugation.Complete cell sheet layer can be fabricated with temperature-responsive culture dishes.

AIM: To evaluate the repeatability and reproducibility of thickness profile measurement of intra-retinal layers determined by an automated algorithm applied to OCT images from Cirrus optical coherence tomography (OCT) instrument.METHODS: In this prospective cross-sectional study,retinal thickness images at 6mm×6mm around fovea were obtained from 86 eyes of 43 normal subjects with Cirrus HD-OCT instrument.The retinal images from patients were analysis by Cirrus automated algorithm GCA software,including ganglion cell layer and inner plexiform layer(GCIPL).During this study,operator A would make 2 times measurements to all patients,after that operator B would make another 2 times measurements by Repeat scan model.All the data,including GCL average thickness,min thickness,12 clock average thickness,2 clock average thickness,4 clock average thickness,6 clock average thickness,8 clock average thickness and 10 clock average thickness were measured by SPSS 15.0 software.Standard deviation (SD),coefficient of variation (CV) and interclass correlation coefficient (ICC) were calculated from the results of three-times tests by different examiner to evaluate the repeatability and from the results of two different examiners to assess the reproducibility.Written informed consent was obtained from each subject prior to any medical procedure.RESULTS: The average GCIPL thickness of OD was 85.12±3.95μm;the minimum average GCIPL thickness was 83.21±4.41μm;the standard deviation of OD in clock map was from 3.79μm to 4.84μm;CV(95%CI) was between 0.26 and 0.93;ICC was from 0.939 to 0.989.The average GCL+IPL thickness of OS was 84.65±8.73μm;the minimum average GCPL thickness was 81±13.08μm;the standard deviation of OS in clock map was from 8.24μm to 10.56μm;CV(95%CI) was between 0.92 and 4.94;ICC was from 0.264 to 0.968.CONCLUSION: Cirrus HD-OCT GCA software with automated algorithm can offer accurate and repeatable thickness profile of OCT retinal image.GCA is a useful and reliable approach to the measurement of intra-retinal layer thickness,which may improve the diagnosis and monitoring of retinal diseases.

AIM:Atherosclerosis primarily involved systemic arteries .Luminal surface , a monolayer of endothelial cells , of artery directly exposes to blood and is susceptible to active substances in the blood .Exosomes contain significantly amount of proteins and RNAs .Ex-osomes can be good and bad for cells , depending on their component .Thus, exosomes may contribute to atherosclerosis by affecting endothelial cells .This study analyzed the relationship of exosome proteins and atherosclerosis .METHODS: Fifty-six patients and healthy subjects were recruited and divided into two comparisons:healthy subjects vs atherosclerosis ( HS vs AS) , and hypertension vs hypertension plus atherosclerosis ( HT vs HT+AS) .Serum exosomes were decoded by protein mass spectrometry .The protein profile and function were analyzed by gene ontology ( GO) .RESULTS:It was found that five child terms repeatedly appeared in “response to stimulus” and “immune system process” of BP of the two categories ( HS vs AS and AS vs HT+AS):“positive regulation of innate immune response”,“immune response-activating signal transduction”,”activation of innate immune response”,“innate immune re-sponse-activating signal transduction” and “innate immune response activating cell surface receptor signaling pathway ”.Two child terms repeatedly showed in “binding” of MF of the two categories:“antigen binding” and “enzyme binding”.Two proteins, PSMA6 and PSMA7, were repeatedly shown in the two categories .CONCLUSION:GO analysis was utilized for structure hierarchy “tree” to illustrate these proteins involved in various terms in BP , CC and MF.The PPI analysis supplied proteins which may play potentially im-portant roles in AS process .Innate immune system and blood coagulation pathway contribute to AS formation .The proteins, PSMA6, PSMA7 and Annexin A2, may can be the new target proteins for prevention and treatment of AS .

AIM:To analyze the proteins included in exosomes derived from blood of patients with hypertension and seek the main pathologi -cal changes in hypertension .METHODS:Forty-seven patients and healthy subjects were recruited and divided into two comparisons :healthy subjects vs atherosclerosis ( HS vs AS) , and atherosclerosis vs hypertension plus atherosclerosis ( AS vs HT+AS) .We extrac-ted exosomes from blood and utilized LC-MS/MS to identify the protein expression .We used GO analysis to established the hierarchy programs of biological process and molecular function .PPI was used to find the proteins related to the terms .RESULTS:It was found that three final child terms repeatedly shown in BP of the two categories ( HS vs AS and AS vs HT+AS):“signal transduction in re-sponse to DNA damage”,“response to zinc ion”, and“platelet aggregation”.It was found that two final child terms in MF of the two categories:“interleukin 2 receptor binding” and“ploy(A) RNA binding”.The proteins, PSMA6, PSMA7 and CA2, were related to the terms in the two categories .CONCLUSION: We discovered that the exosome proteins may indicate the pathological changes in hypertension through the biological processes related with the specific proteins .These specific proteins, such as VCL, PSMA6, DP, AKAP, ATP5B and CA2, can be the new indicators for severity of hypertension and new therapeutic targets .

Objective:To explore the effect of post-pyloric feeding by spiral nasoenteric tubes on the prognosis of critically ill patients with acute gastrointestinal injury (AGI) grade Ⅱ.Methods:A retrospective study was performed to analyze the clinical data of critically ill adult patients with AGI grade Ⅱ, who were enrolled in three randomized controlled trials conducted by Guangdong Provincial People's Hospital for post-pyloric tube placement between April 2012 and May 2019. Data including demographic characteristics, serological indicators of nutrition, the tube tip position confirmed by abdominal X-ray 24 h after tube insertion, and intensive care unit (ICU), 28-day and hospital mortality were collected. Patients were divided into the post-pyloric feeding group and gastric feeding group according to the tube tip position. Propensity score matching method was used to perform 1:1 matching, and the differences of each index between the two groups were compared after matching. Then the influencing factors of P<0.1 were included in multivariate logistic regression analysis to investigate the potential ICU mortality risk factors of critically ill patients with AGI gradeⅡ. Factors with 0.1 level of significance from the univariate analysis were considered in the multivariate analysis. Results:There were 90 patients in post-pyloric feeding group and 90 patients in the gastric feeding group. Demographics and clinical characteristics of study population were well balanced between the two groups after matching. ICU, 28-day and hospital mortality in the post-pyloric feeding group were significantly lower than those in the gastric feeding group (4.4% vs 15.6%, 14.4% vs 27.8%, 6.7% vs 17.8%, all P < 0.05). Multivariate logistic regression analysis indicated that post-pyloric feeding was an independent protective factor [odds ratio ( OR)=0.295, 95% confidence internal (95% CI): 0.091-0.959, P=0.042] and APACHEⅡ score was an independent risk factor ( OR=1.111, 95% CI: 1.025-1.203, P=0.010) for ICU mortality of critically ill patients with AGI gradeⅡ. Conclusions:Post-pyloric feeding for critically ill patients with AGI grade Ⅱ could decrease ICU mortality and is an independent protective factor against mortality.

Proteomics, which has been widely used in life science, is an emerging discipline following genomics. It can help to explore the pathogenic mechanism and early onset marker of Bacillus anthracis, playing an important part in the prevention, diagnosis and treatment of B.anthracis. In this paper,the application of proteomics in the research of B.anthracis is reviewed.

OBJECTIVETo study clinical outcomes of Chinese medidine fumigation and massage therapy for the treatment of knee stability and functional recovery after anterior cruciate ligament reconstruction operation,and to explore the effect on tendon-bone healing.

METHODSTotal 50 patients were divided into two groups: the control group (normal rehabilitation therapy group),the treatment group (Chinese medicine fumigation and manipulation group). There were 25 patients in the control group, including 16 males and 9 females, who were treated with isometric muscle training, with the gradually enlarging amplitude of flexion and progressive loading of bearing training for knee recovery. There were 25 patients in the treatment group, including 15 males and 10 females,who were treated with the conventional rehabilitation therapy combined with Chinese medicine fumigation and massage therapy. The Chinese herbs named as Haitongpi decoction was steamed by a special equipment to fumigate the knee after operation; Based on the biomechanical parameters of the ligament reconstruction, the massage therapy was designed to control the degree of the knee flexion and release the adhesion for early recovery of knee functions. The Lysholm knee function evaluation system was used, and MRI examination was performed to measure the change in width of ligament tunnel in femur and tibia to evaluate the safety and stability of the treatment.

RESULTSLysholm system showed that two groups both had improving results from the 1st month after operation to the 3rd month (treatment group, F=36.54, P<0.05; the control group, F=28.12, P<0.05), and the results of the treatment group was better than that of the control group at the observation point (the 1st month, t=0.105, P<0.05; the 3rd month, t=5.361, P<0.01). There was no difference between the two groups when evaluating the bone and tendon healing 3 and 12 months after operation (P>0.05), indicating that Chinese rehabilitation therapy was a safety treatment without the influence on the loosing of tendon.

CONCLUSIONChinese medicine fumigation and massage therapy can early improve the knee function after the anterior cruciate ligament reconstruction operation without the disturbance of the knee stability.

Adult ; Anterior Cruciate Ligament ; surgery ; Anterior Cruciate Ligament Injuries ; Anterior Cruciate Ligament Reconstruction ; Case-Control Studies ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Fumigation ; Humans ; Knee Injuries ; drug therapy ; physiopathology ; surgery ; therapy ; Knee Joint ; drug effects ; physiopathology ; surgery ; Male ; Massage ; Range of Motion, Articular ; Recovery of Function

OBJECTIVE This study aimed to investigate the influence of IgD on T/B cell activationand construct hIgD-Fc-Ig fusion protein to competitive inhibition IgD binding with IgDR. METHODS T/B cells were sorted by magnetic cell sorting. The differences of mIgD and IgD-R level between different T/B cell subtypes were detected by FCM. Serum IgD level was detected by ELISA. Human IgD-Fc-IgG1- Fc sequence was amplified by cross- PCR and then subcloned into PET28a(+ ) empty vector. After prokaryotic expression through escherichia coli, we obtained the hIgD-Fc-Ig fusion protein by affinity chromatograph. Western blot was used to identify the hIgD- Fc- Ig fusion protein. Human peripheral blood monouclear cells (PBMC) and fibroblast like synoviocytes (FLS) proliferation were detected using a cell counting kit-8 (CCK-8). RESULTS The percentage of CD3+/CD4+, CD3+/IgD+, CD3+/CD4+/IgD+, CD3+/IgD-R+ and CD3+/CD4+/IgD-R+ cells increased significantly in RA patients comparing to healthy people. IgD can stimulate PBMC proliferation. IgD (1, 3, 10, 30 μg·mL-1) stimulate PBMC proliferation significantly after 24 h. We obtained stable and active hIgD-Fc-Ig fusion protein. The hIgD-Fc-Ig fusion protein showed no effect on PBMC proliferation. But it could downregulate human IgD protein promoting proliferation effects in human PBMC. CONCLUSION This result suggests that IgD and IgDR play an important role on T/B cell activation in RA patients and the hIgD-Fc-Ig fusion protein may competitively inhibit IgD's function and may play an therapeutic role in autoimmune diseases.