Abstract: Objective　The method of "molecular docking - molecular dynamics - in vitro pharmacology" was used to screen and optimize small molecule polypeptide compounds that can block PD-1/PD-L1 binding and have high anti-tumor activity. Methods Using the 5-peptide compound database constructed by our team and the three-dimensional structure data of the PD-L1 protein downloaded from the protein database (PDB), the peptide compounds obtained by flexible molecular docking using Molecular Operating Environment software. The molecular dynamics calculation and analysis of the compounds with the highest GWVI/WSA (generalized Born volume integral/weighted surface area) free energy change value (ΔG) were carried out, including the root mean square deviation (RMSD) of the position change of the weight atom and the interaction energy (the sum of Lennard-Jones potential and Coulombic energy). The blocking effect of ligand compounds on PD-1/PD-L1 binding was analyzed by the homogeneous time-resolved fluorescence (HTRF) technique. The co-culture system of Jurkat T lymphocytes and melanoma B16-F10 cells was established to explore the effect of compound ligands on the killing effect of T cells on tumors and the effect on the secretion level of IL-2 in the co-culture supernatant. Results The selected polypeptide compounds were analyzed by molecular dynamics, and the binding of RGGHA to RGGHH and PD-L1 was stable. There are many kinds of interactions between RGGHA and PD-L1，which can compete with PD-1 ligands to bind Asp122, Try123 and Lys124 sites of the PD-L1 protein and block PD-1/PD-L1 signal transduction. HTRF experiments showed that the binding inhibition rate of RGGHA to PD-1 and PD-L1 was 58.38%, and that of RGGHH was 42.73%. In addition, we established a co-culture system of Jurkat T lymphocytes and melanoma B16-F10 cells to explore the immunostimulatory effect and mechanism of peptides. The results show that RGGHA and RGGHH can significantly increase the secretion level of IL-2, improve the killing ability of T cells against tumor cells and activate the tumor immune microenvironment. Conclusions The study found that the polypeptide compounds RGGHA and RGGHH can effectively block the PD-1/PD-L1 interaction and reactivate the anti-cancer immune response, which can be used as lead compounds for new drug development.

【Objective】 To clarify the role and molecular mechanism of Tanshinone ⅡA (TanⅡA) in the pathological integration of granule cells in the dentate gyrus (DG) by using the mouse model of temporal lobe epilepsy (TLE). 【Methods】 Status epilepticus (SE) was induced in the mice with pilocarpine and treated with TanⅡA 5 mg/kg. After two months, Morris water maze was used to examine the spatial learning and memory ability and video surveillance was used to monitor spontaneous seizures. The DG was removed for staining of Timm, Prox-1, DCX and SynⅠ. PTEN, p-AKT, and p-S6 expressions were observed by Western blotting. 【Results】 TanⅡA decreased Timm score, SynⅠ, PSD-95 and pS6 levels, and increased the level of PTEN in the DG, and attenuated the formation of mossy fiber sproutings and basal dendrites of the granule cells. Video surveillance showed that TanⅡA reduced the frequency of Racine’ grade 5 seizures. 【Conclusion】 TanⅡA can effectively attenuate the abnormal integration of the granule cells in the DG by regulating PTEN/AKT/mTOR pathway and thus plays an anti-epileptic role.

Objective:To find out the occurrence of anxiety and depressive symptoms, and the major risk factors, and the participation rate, as well as the experience of medical personnel who are involved in the intervention.Methods:Since January 2018, a pilot intervention had been carried out on pregnant women registered in the antenatal clinic. The Generalized Anxiety Disorder Scale and the Patient Health Questionnaires were used as screening tools for anxiety and depression symptoms, and risk factors were screened too. Interventions were carried out on the psychological moderate and high risk women by obstetric medical staff and mental health personnel. A qualitative interview was conducted on the intervention providers.Results:A total of 9 488 pregnant women were included, and the positive rate of moderate anxiety symptoms was 3.0%, the positive rate of severe anxiety symptoms was 1.4%; the positive rate of moderate depression symptoms was 18.1%, and the positive rate of severe depressive symptoms was 5.2%; the comorbidity rate of anxiety and depression symptoms was 3.4%. The first three risk factors for pregnant women with anxiety symptoms were: once had premenstrual stress symptom, excessive fear of fetal growth, previous abnormal maternal history; the first three risk factors for pregnant women with depressive symptoms: once had premenstrual stress symptom, previous abnormal maternal history, this pregnancy was cherished; the first three risk factors for pregnant women with moderate and above anxiety combined with depression were: once had premenstrual stress symptom, excessive fear of fetal growth, and fear the delivery process is not successful. Among the psychological moderate risk pregnant women, 19.1% participated in the midwife joint counselor clinic, and 1.7% participated in the obstetrician joint psychological specialist nurse clinic, 2.2% of the pregnant women with high risk participated in the psychological multidisciplinary consultation, and 1.7% referred to the psychiatric department. From the interviews, providers believed that it was necessary to further strengthen the ability of psychological intervention capacity, and the psychological screening tools needed to be improved, and the problems sought by pregnant women involved in physical, psychological and social aspects, and the influence of pregnant women's treatment compliance included multiple factors.Conclusions:The psychological health care service during pregnancy was feasible, but the screening scales needed further examination. The mental health care ability of obstetric medical staff needed to be strengthened, and the compliance of pregnant women with mental health services needed to be improved.

A limited number of microRNAs (miRNAs, miRs) have been reported to control postnatal cardiomyocyte proliferation, but their strong regulatory effects suggest a possible therapeutic approach to stimulate regenerative capacity in the diseased myocardium. This study aimed to investigate the miRNAs responsible for postnatal cardiomyocyte proliferation and their downstream targets. Here, we compared miRNA profiles in cardiomyocytes between postnatal day 0 (P0) and day 10 (P10) using miRNA arrays, and found that 21 miRNAs were upregulated at P10, whereas 11 were downregulated. Among them, miR-31a-5p was identified as being able to promote cardiomyocyte proliferation as determined by proliferating cell nuclear antigen (PCNA) expression, double immunofluorescent labeling for α-actinin and 5-ethynyl-2-deoxyuridine (EdU) or Ki-67, and cell number counting, whereas miR-31a-5p inhibition could reduce their levels. RhoBTB1 was identified as a target gene of miR-31a-5p, mediating the regulatory effect of miR-31a-5p in cardiomyocyte proliferation. Importantly, neonatal rats injected with a miR-31a-5p antagomir at day 0 for three consecutive days exhibited reduced expression of markers of cardiomyocyte proliferation including PCNA expression and double immunofluorescent labeling for α-actinin and EdU, Ki-67 or phospho-histone-H3. In conclusion, miR-31a-5p controls postnatal cardiomyocyte proliferation by targeting RhoBTB1, and increasing miR-31a-5p level might be a novel therapeutic strategy for enhancing cardiac reparative processes.
Animals ; Cell Count ; MicroRNAs ; Myocardium ; Myocytes, Cardiac* ; Negotiating ; Proliferating Cell Nuclear Antigen ; Rats

Objective To explore the relationship between achievement motivation,procrastination and positive mental health of college students.Methods A total of 348 students completed a survey that in-cluded Achievement Motivation Scale(AMS),General Procrastination Scale(GPS)and Mental Health Con-tinuum Short Form(MHC-SF).Results (1)The average scores of achievement motivation,procrastination and positive mental health in college students were 3.61±12.05,56.48±10.62 and 3.46±0.82 respectively. The proportion of flourishing student was accounted for 53.16%,44.83% for the moderately mentally healthy and 2.01% for the languishing.(2)College students' achievement motivation was positively correlated with positive mental health(r=0.399,P<0.01),achievement motivation was negatively correlated with procrasti-nation(r=-0.388,P<0.01)and procrastination was negatively correlated with positive mental health(r=-0.267,P<0.01).(3)Procrastination had a partial mediating effect between achievement motivation and positive mental health,and the proportion of mediating effect in total effect was 12.75%.In three dimensions of positive mental health,procrastination also had partial mediating effect between achievement motivation and psychological well-being and emotional well-being respectively,the proportion of mediating effect in total effect were 15.71% and 20.63% respectively.But the mediating effect between achievement motivation and social well-being was not significant.Conclusion College students with higher achievement motivation have higher positive mental health,but the procrastination has indirect negative effect between the two variables,which leads to the negative effect of emotional and psychological well-being in the positive mental health condition.

Objective To investigate the surgical strategy and outcome for occipitocervical extramedullary tumors.Methods 15 patients with occipitocervical extramedullary tumors from January 2007 to July 2010 were reviewed retrospectively.There were 7 males and 8 females with an mean age of 44.6 years (range,21-72 years).All cases were intradural tumors,including 7 cases in dorsolateral,5 cases in lateral side,and 3 cases in ventralis of spinal cord.Functional and neurological statuses were assessed using the Frankel grade and Japanese Orthopaedic Association (JOA) scale.According to Frankel grade system,there were 4 patients with Grade C,8 with Grade D,and 3 with Grade E.The mean preoperative JOA score was 10.5±3.6.Based on the tumor site,there were 5 cases in medulla oblongata-C1 and 10 cases in C1,2.Frankel grade and JOA score were used to evaluate neurological status and general health.The improvement of symptoms and characteristics of surgical strategies were analyzed.Results All cases were followed up for 2-4 years (mean 2.8 years).Complete resection of tumor and good fusion of bone graft were found in radiography postoperatively.The pathological diagnoses included 10 cases of schwannoma,3 cases of meningioma and 2 cases of neurofibroma.There was statistical difference between the preoperative and the final follow-up functional and neurological statuses including Frankel grade and JOA score.At the latest follow-up,2 cases improved from Frankel grade C to D,2from C to E,6 from D to E,but none in 2 cases with grade D.The mean JOA score at final follow-up was 15.6±1.6,and the average improvement rate was 80.7％± 17.4％.The mean preoperative C0-2 angle was 27.0°±4.1 °,and C2-7 angle was 16.8°± 12.7°.C0-2 angle was 27.6°±8.8°,and C2-7 angle was 10.2°±6.8° at the latest follow-up.However,these differences did not reach statistical significance.Conclusion Surgical treatment can effectively maintain or improve neurological function and improve quality of life.Extramedullary tumors can be resected completely by posterior approach,and spinal stabilization can be obtained satisfactorily through selecting appropriate surgical strategies.For medulla oblongata-C1 level,occipitocervical fusion is usually chosen after extirpation of tumor.For C 1,2 level,C1,2 fusion after tumor resection is useful in preventing atlantoaxial instability.

OBJECTIVETo observe the morphological changes during development of the ventricular zone (VZ) and subventricular zone (SVZ) of human fetus and the distribution pattern of neural stem cells in the VA and SVZ.

METHODSHuman fetuses at the gestational ages of 9-11 weeks, 14-16 weeks, 22-24 weeks and 32-36 weeks were collected, and the brain sections of the VZ/SVZ under the frontal lobe were examined for cytoarchitecture and distribution of nestin-positive cells with HE staining, immunohistochemistry or immunofluorescence.

RESULTSThe thickness of VZ underwent no significant changes at the gestational ages of 9-24 weeks (P>0.05) and became obviously thinner at 32 weeks (P<0.05), while the thickness of SVZ increased during 9-24 weeks (P<0.05) without obvious thinning at 32 weeks (P>0.05). VZ was thicker than SVZ at 9-11 weeks but became markedly thinner than SVZ after 14 weeks (P<0.05). The VZ contained denser cells than SVZ and showed a distinct boundary between the VZ and SVZ. Large numbers of nestin-positive cells were detected in the VZ and SVZ, and nestin immunoreactivity was found primarily in the cell processes and occasionally in the soma. Some nestin-positive cells in the SVZ had 1-3 processes. Nestin immunoreactivity in the VZ and SVZ gradually grew weak with development. The cells positive for both nestin and Ki67 were located mainly in the inner zone of the VZ and throughout the SVZ, where some nestin-positive but Ki67-negative cells were also found.

CONCLUSIONThe SVZ fully extends and the neural stem cells in the VZ/SVZ can be morphologically heterogeneous during the development of fetal human brain.

Fetus ; Frontal Lobe ; cytology ; embryology ; metabolism ; Humans ; Nestin ; metabolism ; Neural Stem Cells ; metabolism ; Neurons ; metabolism

OBJECTIVETo investigate the expression of metabotropic glutamate receptor 5 (mGluR5) and its cellular distribution in the frontal cortex, ventricular zone (VZ) and subventricular zone (SVZ) in human fetuses.

METHODSAccording to the gestational age, the collected fetuses were divided into 4 groups, namely 9-11 weeks, 14-16 weeks, 22-24 weeks and 32-36 weeks. Brain tissue blocks including the frontal lobe or VZ/SVZ were prepared into slices, and the expression pattern and cellular distribution of mGluR5 in the frontal cortex and VZ/SVZ were observed by immunohistochemistry or immunofluorescence.

RESULTSmGluR5 immunoreactivity was present in the cell membrane in the frontal cortex, VZ and SVZ from the 9th to 36th weeks and the immunoreactivity in the marginal zone (MZ) and cortical plate (CP) was markedly stronger than that in VZ and SVZ. The cells expressing mGluR5 included neural stem/progenitor cells in the VZ and SVZ, immature neurons in the VZ and MZ, and numerous mature neurons in the CP.

CONCLUSIONmGluR5 is expressed by a variety of cells such as neural stem cells in the frontal cortex, VZ and SVZ in human fetus, suggesting a role of mGluR5 in the development of human cerebral cortex.

Cerebral Cortex ; cytology ; Cerebral Ventricles ; cytology ; metabolism ; Fetus ; cytology ; metabolism ; Frontal Lobe ; cytology ; metabolism ; Humans ; Neural Stem Cells ; cytology ; metabolism ; Receptor, Metabotropic Glutamate 5 ; Receptors, Metabotropic Glutamate ; metabolism

Objective To study the comprehensive therapeutic effect of interventional embolization chemotherapy in advanced cervical cancer. Methods 136 patients with advanced cervical cancer were selected as research object,and they were randomly divided into control group(radiotherapy group) 68 cases and observation group(interventional embolization chemotherapy group) 68 cases,then the total effective rate,incidence of adverse reactions,secondary operation rate and serum CA125 ,SCC and CY211 of two groups before and after the treatment were analyzed and compared. Results The total effective rate( 83. 8% vs 66. 2% ) and secondary operation rate(89.7% vs 48.5% )of observation group was higher than that of control group, incidence of adverse reactions (63.2% vs73.5 % ) was lower than that of control group,serum CA125 [(37. 89 ± 16. 78) vs(52. 36 ± 16. 98) U/ml], SCC[(0. 68 ± 0. 42) vs (1.52±0.36)μg/L]and CY211[(4.86 ±2.21)vs(8.45 ±2.85)ng/L]were all lower than those of control group. ( P ＜0.05 or P ＜0.01) , there were significant differences. Conclusion The comprehensive therapeutic effect of interventional embolization chemotherapy in advanced cervical cancer was better,and it was one of effective methods in advanced cervical cancer.

Objective To explore the images and diagnostic evaluation of X-ray combined with ultrasound in paralytic intestinal obstruction caused by antipsychotic agents. Methods The data of X-ray and ultrasound of 124 cases of clinically diagnosed paralytic intestinal obstruction caused by antipsychotic agents were analyzed retrospectively, as well as their rates of confirmed diagnosis. All the 124 patients were examined by erect abdominal radiography and fluoroscopy,68 of the 124 patients by ultrasound. Results According to their typical images,the rate of X-ray confirmed diagnosis was 87.9%(109/124) with 15 missed diagnosis and the rate of missed diagnosis was 12.1%( 15/124);the rate of ultrasound confirmed diagnosis was 82.4%(56/68) with 12 missed diagnosis and the rate of missed diagnosis was 17.6%(12/68). There was no significant difference between X-ray and ultrasound (P＞ 0.05 ). Compared with X-ray or ultrasound ,X-ray combined with ultrasound in the diagnosis of paralytic intestinal obstruction could increase the rate of confirmed diagnosis (100.0% ,68/68),and decrease the rate of missed diagnosis (0),the difference was statistically significant (P＜ 0.05 ). Conclusions X-ray examination is the first choice to diagnose paralytic intestinal obstruction caused by antipsychotic agents,but ultrasound has the advantage of finding fluid in abdominal cavity. To reduce missed diagnosis, X-ray should be combined with ultrasound.