Objective To investigate the role of hippocampal mu-opioid receptors (MORs) in the formation of seizure susceptibility induced by kainic acid (KA). Methods The SD rats were injected subcutaneously with a convulsive dose of KA (10 mg/kg). Then mini-osmotic pumps were used for intrahippocampal injection with a selective MOR antagonist ?-FNA, and a selective MOR agonist PL017 in these rats. 7 days later, the seizure susceptibility was checked by a subthreshold dose of KA (5 mg/kg). Results All animals showed motor seizures over Stage 4 after a convulsive dose KA injection. 7 days later, 100% animals showed seizures over Stage 4 in KA, CSF + KA and PL017+KA groups after injection of subthreshold dose of KA, but only 28.5% in PL017+KA group. Mean latency of PL017+KA group was (10.1?3.0) minutes, shorter significantly than that of KA and CSF+KA groups (0.010.05). The mean stage of ?-FNA + KA group was 1.7?0.1, being reduced significantly as compared with the stages of other three groups ( P

Objective To establish immortalized neural progenitor cell strain and provide stable cell resource for cell-transplantation and gene therapies. Methods Plasmid pCMVSV40T/PUR containing the simian virus 40 large T antigen gene (SV40Tag) were transfected into the primary cultured neural progenitor cells (NPCs) of newborn rat using lipofectin transfection method. Colonies were isolated by puromycin selection and expanded by many passages. Anti-nestin antibodies were used to identify the cultured cells. The specific molecular marker of neurons and astrocytes were detected using immunocytochemistry method to investigate the capability of differentiation of the transfected cells. The expression of SV40Tag in expanded cell lines was identified by RT-PCR, Southern blot and immunocytochemistry method.Results One anti-puromycin cell clone was obtained, which was microtubule-associated protein-2 (MAP-2) positive cells with the capability of proliferation and could differentiate into MAP-2 or glial fibrillary acidic protein positive cells. The existence of SV40Tag cDNA and the expression of mRNA and protein of SV40Tag were confirmed in transfected cells. The transfected cells were expanded to immortalized cell strain maintained for more than 50 passages, named as immortalized neural progenitor cell (INPC) . INPCs were elliptical or triangular cells with two or three short axons. The population doubling time of INPC was (22.9?2.7)h, subculture, freezing and recovering had no effect on cellular shape and proliferation of INPC. Conclusion Immortalized neural progenitor cell strain was established successfully. It may provide stable cell resource for the basic researches and cell-transplantation therapies with NPC.

Objective To approach the effect of blocking the JAK-STAT signal pathway in the treatment of rat's rheumatoid arthritis.Methods Thirty-six arthritis rat models,which had arthritis index more than 2 scores,were built successfully by Department of Rheumatology and Immunology,the First Affiliated Hospital of China Medical University,and they were divided into model group one and two,low dose of AG490 group,middle dose of AG490 group,high dose of AG490 group and leflunomide group.Besides,6 rats were treated as normal control group.And they were given normal sodium 1 mL,AG490 1、5、10 mg/(kg?d)and leflunomide 10 mg/(kg?d) by intraperitoneal injection or intragastric administration,then all the rats were sacrificed after 44 days.After that,compared the weight,arthritis index,the volume of toe,pathological score,and the change of p-STAT1,p-STAT3.Results The arthritis of model group developed rapidly,and the arthritis index,the volume of toe increased obviously,and the expression of p-STAT1 and p-STAT3 increased and the body weight decreased,all of which had significant meaning compared with that of control group.The body weights of rats in low dose group of AG490 were heavier than that of control group,the symptom of arthrocele in low dose group of AG490 was lighter than that of control group,and could inhibited the activation of JAK-STAT signal pathway in some degree,but the arthritis index numbers and the pathological score had no significant difference from that of model group.The body weights,arthritis index,the volume of toe and pathological score in middle dose group of AG490 improved obviously(P

Objective]To explore the clinical value of the theories in the seven papers of Yunqi.[Method] Analysing the usage of the correlation theories of Yunqi in Cases of Well-known Physicians and its sequel, consulting the commentaries and application of many well-known physicians, this paper summarizes some theories which are applicable to clinic.[Result]In the seven papers of Yunqi, the theory of avoidance of harm by restraining excess and the theory about the inconformity of pulse and disease can exclude false symptoms and infer real pathogenesis.19 questions about pathogenesis can be a reference of clinical diagnosis.The theory of South-North can be used to exclude the impact of climate on the pulse and assist diagnosis. Rationally using the theory of Suiyun contributes to infer etiology and establish therapy.[Conclusion]Cases of Well-known Physicians and its sequel reflect the application and exertion of the theories of Yunqi by ancient physicians. They help to understand the theoretical system and instruct clinical application. In addition to the theory of Suiyun, the seven papers of Yunqi also include a lot of theories which are applicable to clinic.

Objective To investigate the clinical characteristics and therapeutic plan of Miller Fisher syn-drome( MFS) complicated with Guillain-Barrésyndrome( GBS) .Methods The clinical data of one patient of MFS complicated with GBS was reviewed retrospectively,and analysis combined with literature review was given.Results The patient had infectious history before the onset of MFS and GBS.Clinical manifestations included MFS triad,bulbar paralysis,bilateral facial paralysis,protein-cell dissociation of cerebrospinal fluid and neurogenic damages in electro-myography.Plasm exchange and adrenocortical hormones treatment were effective.Conclusion MFS complicated with GBS is an independent disease which is different from MFS or GBS.It has its distinct clinical features which should apply appropriate treatment.

AIM: Screening of phage antibodies against HBsAg from the constructed phage display library and sequencing of the positive clones.METHODS: RNAs were extracted from blood lymphocytes of a HBsAg-immunized volunteer. Then cDNAs were synthesized by RT-PCR. To construct the phage antibody display library, cDNAs of Fd fragments and ? chains of the antibodies were amplified through PCR and were inserted into vector pComb3H. Three rounds panning against coated HBsAg showed the specific enrichment of phage antibody. Positive clones were selected and sequenced.RESULTS: A phage antibody display library was constructed after amplifing of Fd fragments and ? chains of IgG1. Three Fab antibodies from the library were selected and sequenced. The result of sequencing showed that two of the three antibodies had the same Fd fragment and all of the three had the same light chain. CONCLUSION: The phage antibody display library was constructed successfully and three specific antibodies (Fab fragments) have been obtained from it.

Objective To investigate the expressions and correlations of Kiss-1 and matrix metalloproteinases-9 (MMP9) proteins in the colorectal cancer (CRC) tissues of patients with synchronous colorectal liver metastasis (SCRLM),and the association with the clinicopathologic factors and prognosis of patients.Methods The retrospective case-control study was adopted.The clinicopathological data of 96 patients with SCRLM and 69 patients with CRC and no metastasis who were admitted to the Eastern Hepatobiliary Surgery Hospital of the Second Military Medical University from January 2000 to May 2013 were collected.The 96 CRC tissues and 50 adjacent normal tissues (distance from resection margin ≥ 5 cm) were collected from 96 patients with SCRLM,and 69 CRC tissues were collected from 69 patients with CRC and no metastasis.Expressions of Kiss-1 and MMP9 protein were detected by immunohistochemistry (IHC).The follow-up of outpatient examination and telephone interview was performed to detect survival of patients till August 2014.Comparison of count data and correlation between expressions of Kiss-1 or MMP9 protein and clinicopathological factors were analyzed by the chi-square test and Fisher exact probability.Survival curve was drawn using the Kaplan-Meier method,and survival analysis was done using the Log-rank test.Correlation analysis was done by the Pearson correlation.Results Expression of Kiss-1 protein was located in the cytoplasm of tissue cells.The positive expression rates of Kiss-1 protein in CRC tissues of patients with SCRLM and with CRC and no metastasis and in adjacent normal tissues were 24.0％ (23/96),43.5％ (30/69) and 52.0％ (26/50),respectively,with a significant difference among the 3 tissues (x2 =14.307,P ＜ 0.05) and no significant difference between CRC tissues of patients with CRC and no metastasis and patients with SCRLM (x2 =0.845,P ＞ 0.05).The positive expression rate of Kiss-1 protein in CRC tissues of patients with SCRLM was significantly different from that in CRC tissues of patients with CRC and no metastasis and in adjacent normal tissues (x2 =0.702,11.594,P ＜ 0.05).Expression of MMP9 protein was located in the cytoplasm of tissue cells.The positive expression rates of MMP9 protein in CRC tissues of patients with SCRLM and with CRC and no metastasis and in adjacent normal tissues were 67.7 ％ (65/96),62.3 ％ (43/69) and 36.0％ (18/50),respectively,with a significant difference among the 3 tissues (x2=14.203,P ＜0.05) and no significant difference between CRC tissues of patients with CRC and no metastasis and patients with SCRLM (x2=8.038,P ＞ 0.05).The positive expression rate of MMP9 protein in CRC tissues of patients with SCRLM was significantly different from that in CRC tissues of patients with CRC and no metastasis and in adjacent normal tissues (x2 =13.475,13.475,P ＜ 0.05).The positive expression rates of Kiss-1 protein in CRC tissues of patients with SCRLM were 66.7％,21.9％ and 17.6％ in the high-,mederate-and low-differentiated tumor,50.0％,28.6％ and 17.5％ in the muscular layer of tumor invasion,outside of serosa and serosal layer,44.0％ and 16.9％ in patients with and without lymph node metastasis,respectively,showing significant differences among the tumor differentiation degree,depth of tumor invasion and lymph node metastasis (x2=6.546,6.172,7.453,P ＜0.05).The positive expression rates of MMP9 protein in CRC tissues of patients with SCRLM were 25.0％,66.7％ and 76.2％ in the muscular layer of tumor invasion,outside of serosa and serosal layer,44.0％ and 76.1％ in patients with and without lymph node metastasis,respectively,showing significant differences between the depth of tumor invasion and lymph node metastasis (x2 =12.094,8.690,P ＜ 0.05).All the 96 patients with SCRLM were followed up for a median time of 68 months (range,12-176 months).The median overall survival time,median tumor-free survival time,5-year cumulative survival rate and 5-year tumor-free survival rate were 31 months,26 months,69.6％ and 26.1％ in patients with SCRLM and positive expression of Kiss-1 protein and 26 months,19 months,24.7％ and 12.3％ in patients with SCRLM and negative expression of Kiss-1 protein,respectively,showing significant differences between the overall survival and tumor-free survival (x2=16.578,14.436,P ＜ 0.05).The median overall survival time,median tumor-free survival time,5-year cumulative survival rate and 5-year tumor-free survival rate were 31 months,19 months,24.6％ and 12.3％ in patients with SCRLM and positive expression of MMP9 protein and 28 months,16 months,58.1％ and 22.6％ in patients with SCRLM and negative expression of MMP9 protein,respectively,showing significant differences between the overall survival and tumor-free survival (x2=14.073,8.532,P ＜0.05).Of 96 patients with SCRLM,there were 23 patients with positive expression of Kiss-1 protein (10 with positive expression of MMP9 protein and 13 with negative expression of MMP9 protein) and 73 with negative expression of Kiss-1 protein (55 with positive expression of MMP9 protein and 18 with negative expression of MMP9 protein),with a negative correlation between expressions of Kiss-1 protein and MMP9 protein (r =-0.291,P ＜ 0.05).Conclusions The reduced expression of Kiss-1 protein and elevated expression of MMP9 protein are closely associated with invasion and metastasis of CRC and prognosis of patients.A combination detection of Kiss-1 and MMP9 proteins is expected to become a marker for predicting the prognosis of patients with SCRLM based on the negative correlation between them.

Objective : To investigate the prevalence of hepatitis C virus (HCV) and syphilis among drug users (DUs) in Inner Mongolia Autonomous Region. Methods : DUs' epidemiological data were collected from 5 national AIDS surveillance sentinels in Inner Mongolia Autonomous Region in 2021 through the National HIV/AIDS Sentinel Surveillance Data Management System, and sexual behaviors, DU behaviors, HIV testing and behavioral interventions were descriptively analyzed. The factors affecting the HCV and syphilis infections were identified among DUs using a multivariable logistic regression model. Results : A total of 2 019 DUs were enrolled, with a mean age of (43.32±9.88) years. Among the participants, there were 1 958 males (96.98%), 1 830 with local registered residence (90.64%), 1 038 with marriage/cohabitation (51.41%), 1 410 with an educational level of junior high school and above (69.84%), and 1 884 with Han Ethnicity (93.36%). There were 1 605 DUs with use of conventional drugs (79.49%), 283 with use of new drugs (14.02%) and 131 with use of mixed drugs (6.49%), and there were 340 DUs with injection drug use (16.84%), including 41 that shared the needles (12.06%). There were 460 DUs with commercial sexual behaviors in the past year (22.78%). The percentage of HCV and syphilis positive was 23.63% and 3.81%, respectively, while no HIV positive case was detected. In addition, multivariable logistic regression analysis showed that age (40 years- , OR=3.747, 95%CI: 2.161-6.497; 50 years and older, OR=5.302, 95%CI: 3.064-9.177), local registered residence (OR=3.409, 95%CI: 1.935-6.008), use of conventional drugs (OR=1.477, 95%CI: 1.035-2.108), and injection drugs (OR=2.700, 95%CI: 2.067-3.527) caused an increased risk of HCV infections among DUs, and divorced/widowed (OR=3.413, 95%CI: 1.463-7.962) and non-local registered DUs (OR=2.404, 95%CI: 1.304-4.433) had an increased risk of syphilis infections. Conclusions Middle-aged men are predominant DUs in national AIDS surveillance sentinels in Inner Mongolia Autonomous Region, and the prevalence of HCV and syphilis infections correlates with age, household registration, marital status, type of drugs used and route of drug use.

Objective: To investigate the prevalence of hepatitis C virus (HCV) infections among five high-risk populations in Inner Mongolia Autonomous Region, so as to provide insights into improvements in the control strategy for hepatitis C. Methods: The detection of anti-HCV antibody was collected from patients receiving renal dialysis, patients receiving invasive diagnosis and treatment in hospitals, physical examination populations, unpaid blood donors and subjects admitted to family planning clinics in national hepatitis C surveillance sentinels in Inner Mongolia Autonomous Region from 2013 to 2021, and the year-, gender- and age-specific prevalence of anti-HCV antibody was analyzed. Results: The mean prevalence of anti-HCV antibody was 2.19%, 1.81%, 0.05%, 0.10% and 0.03% among 3 600 patients receiving renal dialysis, 3 600 patients receiving invasive diagnosis and treatment in hospitals, 18 000 physical examination populations, 18 000 volunteer blood donors and 3 600 subjects admitted to family planning clinics, respectively. The prevalence of anti-HCV antibody appeared a tendency towards a decline among patients receiving renal dialysis (χ2trend=49.065, P<0.001) and volunteer blood donors (χ2trend=11.419, P=0.001). The prevalence of anti-HCV antibody was higher among male patients receiving invasive diagnosis and treatment in hospitals than among females (2.34% vs. 1.36%; χ2=4.826, P=0.028), and no gender-specific prevalence of anti-HCV antibody was seen among other four high-risk populations (all P>0.05). The highest prevalence of anti-HCV antibody was detected among patients receiving renal dialysis (3.30%) and patients receiving invasive diagnosis and treatment in hospitals at ages of 50 to 59 years (3.35%), while the highest prevalence was found among physical examination populations at ages of 60 years and greater (0.18%). Conclusion The prevalence of anti-HCV antibody was high among patients receiving renal dialysis and patients receiving invasive diagnosis and treatment in hospitals and low among physical examination populations, volunteer blood donors and subjects admitted to family planning clinics in Inner Mongolia Autonomous Region from 2013 to 2021. Periodical monitoring of anti-HCV antibody is recommended among the elderly.

The purpose of the study is to construct R8 peptide (RRRRRRRR) and pH sensitive polyethylene glycols (PEG) co-modified liposomes (Cl-Lip) and utilize them in breast cancer treatment. The co-modified liposomes were prepared with soybean phospholipid, cholesterol, DSPE-PEG2K-R8 and PEG5K-Hz-PE (pH sensitive PEG). The size and zeta potential of Cl-Lip were also characterized. The in vitro experiment demonstrated that the Cl-Lip had high serum stability in 50% fetal bovine serum. The cellular uptake of Cl-Lip under different pre-incubated conditions was evaluated on 4T1 cells. And the endocytosis pathway, lysosome escape ability and tumor spheroid penetration ability were also evaluated. The results showed the particle size of the Cl-Lip was (110.4 ± 5.2) nm, PDI of the Cl-Lip was 0.207 ± 0.039 and zeta potential of the Cl-Lip was (-3.46 ± 0.05) mV. The cellular uptake of Cl-Lip on 4T1 cells was pH sensitive, as the cellular uptake of Cl-Lip pre-incubated in pH 6.0 was higher than that of pH 7.4 under each time point. The main endocytosis pathways of Cl-Lip under pH 6.0 were micropinocytosis and energy-dependent pathway. At the same time, the Cl-Lip with pre-incubation in pH 6.0 had high lysosome escape ability and high tumor spheroid penetration ability. All the above results demonstrated that the Cl-Lip we constructed had high pH sensitivity and is a promising drug delivery system.
Animals ; Cell Line, Tumor ; Cell-Penetrating Peptides ; chemical synthesis ; chemistry ; Cholesterol ; chemistry ; Drug Delivery Systems ; Liposomes ; Mice ; Oligopeptides ; chemical synthesis ; chemistry ; Particle Size ; Phospholipids ; chemistry ; Polyethylene Glycols