Objective To explore the functional expression and temporality of MDR1 gene in bone marrow of rabbits after autologous bone marrow transplantation with MDR1 transferred bone marrow mononuclear cells. Methods The supernatant of the amphotropic virus producer cell line PA317-HaMDR1/A was collected and concentrated to cocultivate with the bone marrow mononuclear cells of the rabbits. After large dose of chemotherapy with cyclophosphamide,the transferred cells were autotransplanted into the bone marrow. The integration,transfection rate and physiological function of MDR1 gene were tested by PCR,SP immunohistochemical method and daunorubicin (DNR) extrusion test respectively. Results After autologous bone marrow transplantation had been executed for 1-4 months,the integration of MDR1 gene in genome of bone marrow mononuclear cells was detected by PCR,and the expression rates of P-gp in cells tested by SP immunohistochemical method were 9.5%,8.5%,6.0% and 3.5% respectively. The physiological function of MDR1 gene in bone marrow cells was proved by DNR extrusion test. Conclusion After the autotransplantation with bone marrow mononuclear cells transferred by MDR1 gene,the MDR1 gene can implant into the bone marrow of rabbits and has expressed functionally for 4 months,which has provided a basis for further research on chemoprotection experiment of the MDR1 gene transferred into the bone marrow cells.

Based on their own characteristics and learning purposes of the child neurologic training doctors and to full use of the advantage of our hospital in faculties, facilities and patients, the plan was designed and focused on better understanding of clinical related basic theoretical knowledge, improving the ability of clinical research and thesis writing, strengthening communication skills between patients and doctors, and medical ethics training and health regulations learning. The training doctors achieved good results in the aspects of medical skills, research ability and comprehensive professional quality.

Objective:To observe the levels of lymphocyte subgroups in children with severe pneumonia and non -severe pneumonia , and to investigate the change of celluer immune funciton of patients , moreover provide available reference for treatment.Methods:Flow cytometry instrument was used to detect peripheral blood CD 3+T, CD3+CD4+T, CD3+CD8+T, CD19+B lymphocytes and NK cells in 27 children with severe pneumonia and 28 children with non-severe pneumonia.In addition 20 heathy children were selected as the controls .Results:Compared with the heathy control group ,the levels of CD3+T,CD3+CD4+T,CD3+CD8+T lymphocytes and NK cells in children with severe pneumonia decreased significantly (P<0.01,P<0.05,P<0.01,P<0.05),while CD4/CD8 ratio,CD19+B cells were significantly increased (P<0.05,P<0.01).The level of CD3+CD8+T lymphocytes in children with non-severe pneumonia significantly lower than the heathy control group (P<0.01),but the level of CD19+B cells in children with non-severe pneumonia significantly higher than the heathy control group (P<0.01).Compared with non-severe pneumonia group.The levels of CD3+and CD3+CD8+T lymphocytes in children with severe pneumonia decreased significantly (P<0.01),however CD19+B cells were significantly increased ( P<0.01 ) .Conclusion: Immune dysfunction exists in both of children with severe pneumonia and non-severe pneumonia.Children with severe pneumonia has more serious damage in inmmunity .This provides certain theoretical basis for clinical e-valuation and treatment .

Objective To investigate the relationship between peripheral blood circulating follicular helper T cells and antibody-secreting B cells in patients with ankylosing spondylitis (AS). The role of circulating follicular helper T cells and antibody-secreting B cells in the pathogenesis of AS were explored. Methods Flow cytometry was used to detect the levels of peripheral blood CD4 +CXCR5+ T (cTfh), CD4+CXCR5+PD-1+ T, CD4+CXCR5+ICOS+ T, CD19+ B, CD19+CD38+ B cells in 59 patients with AS (including 36 cases of active AS patients and 23 cases of inactive AS patients). In addition, twenty healthy persons were selected as the controls. Data analysis were performed by independent-sample t test, One way ANOVA analysis, Pearson's and Spearman's correlation test. Results The percentages of cTfh [(26.8 ±10.4)%], CD4+CXCR5+PD-1+T [(12.1±14.0)%], CD4+CXCR5+ICOS+T [(13.6±9.5)%], CD19+CD38+B [(80.7±13.0)%] in the peripheral blood of AS group were significantly higher than healthy controls [(15.6 ±4.5)%, (6.4 ±2.4)%, (9.4 ± 4.5)%, (68.2±13.0)%] (t=6.663, P<0.01; t=2.999, P<0.01; t=2.573, P<0.05; t=2.712, P<0.01). The percentages of cTfh in the active AS group [(30.2 ±11.0)%] were significantly higher than those in the inactive AS group [(21.4±6.5)%] and HC (t=3.444, P<0.01;t=7.004, P<0.01). The percentage of CD19+CD38+B[(85.1±10.0)%] in the peripheral blood of active AS group was significantly higher than that of the inactive AS group [(73.8 ±14.2)%] and HC (t=3.561, P<0.01;t=5.410, P<0.01). The relationship between Bath AS disease activity index (BASDAI) and the percentage of cTfh, CD19+CD38+B was a positive correlation (r=0.442, P<0.01; r=0.405, P=0.001), and significant positive correlation was observed between the percentages of cTfh and CD19+CD38+B cells (r=0.420, P=0.001). Conclusion CD4+CXCR5+cTfh cells are significantly increased in peripheral blood in AS patients with aberrant CD19+CD38+ antibody-secreting B cells, suggesting that cTfh and CD19+CD38+antibody-secreting B cells may play an important role in the pathogenesis of AS .

Objective To construct a recombinant adenovirus encoding ?-galactosidase gene(?-gal),which may be used to treat the lactose intolerance caused by lactase deficiency,with modified AdEasy system.Methods The sequence fragment of ?-gal was cloned into the shuttle plasmid pAdTrack-CMV,and the homo-logous recombination was completated with backbone plasmid pAdEasy1 in the E.coli BJ5183 to construct the recombinant adenoviral plasmid.The adenoviruses were packaged and amplified in the HEK293 cells mediated by liposome.The viral titre was measured by limiting dilution assay and the transfection efficiency was observed by X-gal histochemical staining.Results The recombinant adenovirus with ?-gal was constructed successfully and the viral titre was(2.5~4.0)?1011EFU/ml.More than 70% HEK293 cells were transfected when the MOI was 10.Moreover,more than 65% cells were blue after X-gal histochemical staining.Conclusion Recombinant adenovirus carrying ?-gal with high titre and efficient transfection is constructed successfully by AdEasy system,which supplies valuable experimental basis for the gene therapy to the lactose intolerance caused by lactase deficiency.

Objective:To explore CT and MRI features of the endolymphatic sac tumor (ELST).Methods:The CT and MRI morphology confirmed by surgical pathology for 19 patients with ELST were retrospectively analyzed from June 2011 to May 2019 in Eye & ENT Hospital of Fudan University. The features of CT and MRI included location, size, adjacent structures invasion, CT values, bone destruction, features of T 1WI and T 2WI, enhancement distribution characteristics, dynamic enhancement curve morphology, DWI signal characteristics. The ADC values of the lesions and ipsilateral medial pterygoid muscles were compared using a paired t test. Results:Nineteen ELST patients (one with bilateral diseases) were included. Totally 20 ears (right 9 and left 11) of 13 females and 6 males were studied. The masses with slightly high-density and obscure boundary were located around the vestibular aqueduct at the posterior edge of the petrosal bone. Bone destruction involved mastoid process of the middle ear (16 ears), jugular foramen (11 ears), semicircular canal (10 ears), facial nerve canal (7 ears) and internal auditory canal (9 ears). A large amount of residual bone could be found in the interior of nineteen masses. The CT value was (78.6±21.9) HU. The lesion showed central iso-intensity and peripheral hyperintensity on T 1WI and T 2WI in 16 ears, while no obvious hyperintensity on T 1WI in the other 4 ears. The hyperintensity on T 1WI was around the margin of the lesion in 10 ears, situated at lateral side in 5 ears and all over the lesion in 1 ear. Flow voids signals could be seen in 9 ears as well. Liquid-liquid plane was seen on T 2WI in 2 ears. The solid mass portion which showed iso-intensity on both T 1WI and T 2WI presented marked enhancement on contrast-enhanced T 1WI, while other part of the mass no enhancement. DWI of 14 ears illustrates no evidence of restricted diffusion, and the ADC value [(1.25±0.08)×10 -3 mm 2/s] was slightly higher than that of the medial pterygoid muscles ( t=4.437, P=0.001). The style of time-signal intensity curves of the dynamic contrast-enhanced MRI was rapidly ascending followed by descending curves in 2 ears. Conclusion:Imaging findings of ELST have some characteristics, including located around the vestibular aqueduct at the posterior edge of the petrosal bone, bone destruction, peripheral hyperintensity on T 1WI and no restricted diffusion, which is helpful for its diagnosis.

The clinical thoughts of interns have some features such as insufficiency of initiative and enthusiasm and the absence of systemic. It is necessary to develop their good habits of active and diligent thinking as well as comprehensive thinking ability. Clinical teaching rounds, small lectures,discussion of complicated cases and interactive teaching are the specific measures for our medical interns to improve their ability of clinical thoughts effectifally.

BACKGROUND:Polyvinyl alcohol (PVA) hydrogel with similar porous structure and mechanical properties to the natural cartilage is very suitable for the repair of articular cartilage. However, the pure PVA hydrogel after lyophilization wil be accompanied by the shrinkage of the polymer network and the col apse of the pores, leading to the inhomogeneous performance of the material even in the state of re-swel ing. Addition of the active polymer wil increase the cel adhesion ability of PVA hydrogel. OBJECTIVE:To construct PVA/lota-carrageenan (l-CA) composite materials with different mass fractions of l-CA and evaluate the biocompatibility with vascular endothelial cel s. METHODS:PVA/l-CA composite films with different contents of l-CA were fabricated and then co-cultured with vascular endothelial cel s. Attachment, proliferation and morphological changes of vascular endothelial cel s on the composite were observed by scanning electron microscope and MTT assay to evaluate its biocompatibility. PVA/l-CA three-dimensional scaffold with different contents of l-CA were constructed, and hemolysis experiment was conducted according to the biological evaluation standards of medical devices, and the porosity and pore size were observed using scanning electron microscope. RESULTS AND CONCLUSION:In vitro experimental results showed that the addition of l-CA could significantly increase the biological activity of PVA hydrogel, and promote the cel attachment and proliferation on the scaffold. The hemolysis rate of each experimental group was less than 5%(the accepted safety standard), suggesting that the composite materials were in accordance with the standard of medical devices for hemolysis experiment. These findings indicate that the composite scaffolds with 20%-30%l-CA possess the pore size suitable for cel growth and proliferation and the porosity beneficial for transportation of nutrients and metabolites, which can serve as an excel ent scaffold for tissue engineering.

Objective:To explore the chain mediating effect of depression and rumination between negative life events and non-suicidal self-injury(NSSI).Methods:Adolescents self-harm scale was used to investigate 1 000 students in a vocational school of Guizhou Province. A total of 333 vocational students with NSSI were screened through, and further investigated by adolescent self-rating life events check list (ASLEC), the center for epidemiological studies depression scale (CES-D), and ruminative Response Scale (RRS). SPSS 25.0 and Mplus 8.1 software were used for data analysis, and the bias corrected percentile Bootstrap method was used to test the mediating effect.Results:(1)The scores for negative life events, depression, rumination and NSSI were 67.00 (54.00, 79.00), 47.00 (39.00, 53.50), 64.00 (54.00, 76.00) and 7.00 (2.00, 10.00) respectively.(2)Negative life events were positively correlated with depression ( r=0.29, P<0.01), rumination ( r=0.40, P<0.01), and NSSI ( r=0.18, P<0.01). (3)The mediating effect size of depression between negative life events and NSSI was 0.07, accounting for 38.89% of the total indirect effect.The mediating effect size of rumination between negative life events and NSSI was 0.06, accounting for 33.33% of the total indirect effect.The chain mediating effect of depression and rumination between negative life events and NSSI was 0.05, accounting for 27.78% of the total indirect effect. Conclusion:Negative life events can indirectly affect NSSI of vocational education students through depression and rumination, as well as the chain mediating path.

Spinal muscular atrophy(SMA) is a serious neuromuscular degenerative disease that severely impairs the quality of life for patients and entire families.The emergence of disease-modifying treatments such as nusinersen and risdiplam has gradually changed the natural course of SMA patients.It is particularly important to include the activities of daily living(ADL) ability reported by patients or caregivers in the comprehensive assessment of SMA patients.There are many ADL-related assessment tools, and studies have found that disease-modifying treatments can somewhat improve the ADL ability of SMA children.This article reviews the progress on the effect of disease-modifying treatments on ADL in SMA patients, which can provide a reference for exploring more comprehensive and effective assessment tools and treatment decision-making in subsequent clinical practice.