Objective To explore the mechanism of immune tolerance in chronic B hepatitis, the effects of HBV infection on the functions of macrophages and related mechanism of signal transduction by transfection in vitro were studied. Methods Peritoneal macrophages of mice were isolated regularly and cultured, transfected transiently with pcDNA3-HBV or pcDNA3 plasmid DNA and cultured under the stimulation of LPS. After 72 h, RT-PCR was performed to detect the expression of PreS1, TNF-? or IL-1? mRNA. To detect the expression of NF-?B RelA protein by FCM, and the level of nitric oxide in cultural supernatant was measured with Griess reaction. Results After being transfected with pcDNA3-HBV,peritoneal macrophages had the expression of PreS1 mRNA, but have lower level of TNF-?、IL-1? mRNA and transcriptional factor NF-?B, compared with pcDNA3-transfected control group; the level of nitric oxide in pcDNA3-HBV group was also decreased. Conclusions Transient transfection of pcDNA3-HBV could decrease the function of macrophages directly by inhibiting NF-?B activity and effector molecules production, which may be one of the mechanisms of immune tolerance in chronic B hepatitis.

Objective To study the effect of erythropoietin（EPO） on PI3-K/Akt signal pathway of myocardial ischemia-reperfusion injury（I/R） in immature rabbits. Methods Thirty immature rabbits were randomly divided into two groups （n = 15/greup ）, The rabbits were intraperi- toneally injected with 2 ml of normal saline in control group, or 5 000 U/kg of EPO in treated group 24 h before I/R. The Langendorff isolat- ed perfused heart model was established, and the myocardial expression of PI3K, AKT and Caspase 9 were examined by immunohistochem- istry in 2 h after repeffusion. To monitor the myocardial apoptosis,we performed TUNEL in 2 h after reperfusion. Results The expression of myocardial PI3K and AKT in EPO group was significantly higher in EPO-treated group than those in control group. In EPO-treated group, the expression of myocardial caspase 9 was significantly lower than that in control group. The average optical density of myocardial PI3K, AKT and caspase 9 were significantly different between both groups （P 〈 0.01 ）.The number of apoptotic cell was significantly fewer in EPO- treated group than that in control group. The myocardial apoptosis index （AI）was significantly different between two groups （P 〈 0.01 ）. Conclusion EPO pretreatment can influence the PI3-K/Akt signal pathway and decrease the myocardial apoptosis,and significantly reduce the myocardial ischemia-reperfusion（I/R） injury in immature rabbits.

Objective:To analyze the benefit incidence and evaluate the implementation effects on catastrophic disease insurance under the New Rural Cooperative Medical System ( NRCMS) in Jilin Province and put forward sug-gestions to perfect the catastrophic disease insurance under NRCMS. Method: In-depth interviews with related staff were conducted to gain an understanding of the operating status of catastrophic disease insurance under NRCMS;cat-astrophic disease patient data of Jilin Province from 2013 were collected, with a comparative analysis of implementa-tion effects among nine districts;the benefit incidence was analyzed by means of the Sperman rank correlation analy-sis; Results: The policy of catastrophic disease insurance under the NRCMS exhibited sound implementation effects and played an important role in reducing the economic burden of catastrophic disease patients. The average individual expenditure decreased by 4336. 52 yuan, while the reimbursement rate increased by 11. 15% after NRCMS cata-strophic disease insurance reimbursements. Additionally, statistical analysis results showed that there was a weak cor-relation between benefit incidence and economic level, the benefit equality of catastrophic disease insurance under NRCMS was relatively good, but the benefit level of rich patients was slightly higher. Conclusion:The reimbursement scheme of catastrophic disease insurance under NRCMS should be refined and improved, and the benefit fairness of different districts must be ensured, thus achieving the goal of relieving the“poverty caused by illness” phenomenon.

Objective To explore the roles of Apelin in the development of gastric carcinoma (GC) and interrelation with MVD. Methods Immunohistochemical streptacidin/peroxidase (SP) technique was adopted to examine the expressions of Apelin in 20 cases of normal gastric mucosa, 20 cases of atypical hyperplasia (Dys) and 60 cases of gastric carcinoma, and to examine MVD by CD34 staining. Results With the development of GC, the positive rate of Apelin was increased gradually, namely 5.0 %(1/20) in normal gastric mucosa group, 15.0 %(3/20) in atypical hyperplasia group and 83.3 %(50/60) in gastric carcinoma. Expression of Apelin was correlated with TNM stage and differentiation. MVD in the Apelin positive group (47.01±11.66) was significantly higher than that in the negative group(38.90±10.21); MVD of the Apelin positive group expression was correlated with the histologic stage, serosa infiltration and lymph node metastasis (P < 0.05). Conclusion Apelin may contribute to the development of GC by activating tumour neoangiogenesis.

AIM: To explore the relationship between caspase activation and the evasion of Legionella from macrophage elimination through a Legionella-infected macrophage model. METHODS: After infected by Legionella, the activity of caspase 3 in macrophages was analyzed by confocal microscopy as well as fluorescence reader. Growth and replication of Legionella in macrophage was assayed. Replication of Legionella was analyzed again to see the effect of caspase 3 inhibition on the growth of Legionella after use of caspase 3 inhibitor. RESULTS: Both confocal microscopy and caspase 3 fluorescent substrate analysis showed that Legionella virulent strain had powerful capability of activating caspase 3 while the mutant non-virulent strain did not have this capability. The virulent strain highly replicated in macrophages and the replication was significantly inhibited by caspase 3 inhibitor. CONCLUSION: Our results indicate that the intracellular caspase 3 is activated shortly after infection by Legionella virulent strain. The evasion of Legionella from the elimination of macrophages may be mediated by caspase 3 activation to a great degree.

BACKGROUND: As a candidate gene of congenital heart disease, ACTC1 gene is related to congenital atrial septal defect inhumans.OBJECTIVE: To perform a mutation screen of ACTC1 gene in 110 nuclear families of congenital heart disease.METHODS: A case-control study was conducted based on 110 nuclear families of congenital heart disease and 300 normalhuman beings with no reported cardiac malformation. Six fragments in the coding region of ACTC1 gene was amplified by PCR invitro using five primers pairs. PCR products were screened for gene mutations.RESULTS AND CONCLUSION: A novel G-to-A variant was found at the third nucleotide of the intron downstream from exon 5.This mutation existed in a 5-year-old female with an isolated ventricular septal defect and her 30-year-old father, who had noreported cardiac anomalies. This mutation was not detected in 300 normal controls. These findings indicate that the mutation maybe related with congenital ventricular septal defects in humans.

Objective To investigate the effects of protocadherin 20 (PCDH20) in the spinal cord in the development of bone cancer pain (BCP) in rats.Methods Thirty-six SPF female Sprague-Dawley rats,weighing 180-200 g,were randomly divided into 4 groups (n =9 each):sham operation group (group S),BCP group,lentivirus control group (group LC) and PCDH20 siRNA lentivirus group (group P).Control lentivirus and lentivirus containing PCDH20 siRNA 4 μl were injected into the ipsilateral spinal cord in groups LC and P,respectively.One week later,BCP was induced by injection of Walker 256 breast cancer cells into the upper segment of bone marrow of right tibia.The mechanical paw withdrawal threshold (MWT) was measured at 1 day before injection of lentivirus (T1),1 day before BCP (T2),and 7,14 and 21 days after BCP (T3-5).Three rats in each group were sacrificed after measurement of the MWT at 21 day after BCP and the tibia on the operated side was obtained for examination of invasion of the cancer cells with light microscope.The spinal cord was removed for determination of the expression of PCDH20 and postsynaptic density 95 (PSD95) protein (by Western blot) and mRNA (by RTPCR).Results In groups BCP,LC and P,the cancer cells grew out of the bone and destroyed the cortical bone seriously.Compared with group S,the MWT was significantly decreased at T3-5 in groups BCP,LC and P,the expression of PCDH20 and PSD95 protein and mRNA was up-regulated in groups BCP and LC,and the expression of PCDH20 was up-regulated in group P (P ＜ 0.05).Compared with BCP group,no significant change was found in the MWT and expression of PCDH20 and PSD95 protein and mRNA in group LC (P ＞ 0.05),and the MWT was significantly increased at T4,5 and the expression of PCDH20 and PSD95 protein and mRNA was down-regulated in group P (P ＜ 0.05).Conclusion PCDH20 is involved in the development of BCP through regulating the expression of PSD95 in the spinal cord and adjusting the function of excitatory synapse in rats.

Objective To dynamically monitor the base excess(BE) in traumatic brain injury(TBI) patients within 3 d after admission,and to assess the impact of the early BE on prognosis.Methods Blood BE was monitored for 3 d in 56 TBI patients.Patients were classified into mild group(15 patients),moderate group (22 patients) and severe group (19 patients) according to the scores of Glasgow coma scale(GCS).Patients were classified into survival group(42 patients) and dead group(14 patients) according to prognosis.Patients were classified into high BE group (35 patients,BE≥-8 mmol/L) and low BE group (21 patients,BE ＜-8 mmol/L).The relations among BE,degree of injury and prognosis were analyzed.Results The level of BE in mild group,moderate group and severe group was increased after treatment for 1,2,3 d than that before treatment.The level of BE was consistent with the degree of injury [mild group:(-3.02 ± 0.21)mmol/L; moderate group:(-8.49 ± 1.44) mmol/L;severe group:(-9.64 ± 1.19) mmol/L].The level of BE in mild group and severe group had significant difference than that in moderate group (P＜ 0.01).The level of BE in dead group before treatment and after treatment for 1,2,3 d was significantly lower than that in survival group [(-11.97 ±2.13) mmol/L vs.(-6.29 ± 1.16) mmol/L,(-9.84 ± 1.33) mmol/L vs.(-4.89 ± 1.78)mmol/L,(-8.78 ± 2.01) mmol/L vs.(-3.61 ± 1.43) mmol/L,(-7.84 ± 1.42) mmol/L vs.(-3.10 ±0.98)mmol/L] (P ＜0.01).The scores of APACHE Ⅱ before treatment and fatality rate in low BE group were significantly higher than those in high BE group [(24.84 ± 3.68) scores vs.(16.27 ± 2.21) scores,52.4％ (11/21) vs.8.6％(3/35)] (P ＜ 0.01).The scores of GCS before treatment in low BE group was significantly higher than that in high BE group [(7.56 ± 3.09) scores vs.(10.51 ± 2.43) scores](P ＜ 0.01).Conclusion The level of early BE is a good factor on evaluating the condition and prognosis in TBI patients.

AIM: To explore the effect of TNF-related apoptosis inducing ligand (TRAIL), a new apoptotic inducing molecule on the biological activity of hepatocarcinoma cell line. METHODS: The expression of membrane binding TRAIL on HepG2 cells was detected by immuno-cytochemistry. Quantity of secretory TRAIL was assayed by ELISA method. The cytotoxicity and apoptosis induced by TRAIL was detected by MTT and TUNEL method, respectively. The telomerase activity of HepG2 cells was detected by TRAP-PCR assay kit. The expression of hTERT, the catalytic subunit of telomerase, was detected by FCM. RESULTS: TRAIL was constitutively expressed on the membrane of HepG2 cell line. Soluble TRAIL was also expressed to a certain degree. Cytotoxicity assay showed that TRAIL significantly inhibited the growth of hepatocarcinoma cells. TUNEL assay indicated that TRAIL induced apoptosis in hepatocarcinoma cells. Detection of telomerase activity showed that TRAIL inhibited telomerase activity and the expression of telomerase catalytic subunit. CONCLUSION: TRAIL is an effective molecule to inhibit the growth of hepatocarcinoma through multiple pathways, such as inducing apoptosis and inhibiting the activity of telomerase.

Objective:To compare the effect of O-arm assisted and free-hand pedicle screw placement in the treatment of AO type C thoracolumbar fracture.Methods:A retrospective cohort study was used to analyze the clinical data of 34 patients with type C thoracolumbar fracture admitted to Henan Provincial People′s Hospital from January 2018 to June 2021, including 23 males and 11 females; aged 42-63 years [(50.4±7.4)years]. The fracture was located at T 11 in 4 patients, T 12 in 10, L 1 in 12, L 2 in 6, T 11~12/L 1 in 1 and T 12/L 1 in 1. Posterior reduction and internal fixation was carried out for all patients, of whom 18 were treated with O-arm assisted pedicle screw placement (navigation group) and 16 with free-hand pedicle screw placement (free-hand group). The operation time, single screw placement time, intraoperative bleeding volume, operation mode and screw placement accuracy were compared between the two groups. The kyphotic Cobb angle, visual analogue score (VAS) and American Spinal Injury Association (ASIA) score were compared between the two groups before operation, at 1 week after operation, at 3 months after operation and at the last follow-up. Postoperative complications were observed. Results:All patients were followed up for 12-29 months [(16.8±6.1)months]. There was no significant difference between the two groups in the operation time, intraoperative bleeding volume and operation mode (all P>0.05). The single screw placement time was (9.4±1.6)minutes in navigation group, but was (10.8±1.5)minutes in free-hand group ( P<0.05). The screw placement accuracy was 97.4% in navigation group, but was 81.5% in free-hand group ( P<0.01). The kyphotic Cobb angle and VAS had no significant differences between the two groups before operation (all P>0.05). The kyphotic Cobb angle in navigation group and free-hand group was (4.3±1.1)° and (5.9±1.1)° at 1 week after operation, (4.4±1.2)° and (5.7±1.3)° at 3 months after operation, and (4.4±1.2)° and (6.8±0.9)° at the last follow-up, decreased significantly from that before operation [(21.8±3.1)°, (22.2±3.2)°] (all P<0.01). The kyphotic Cobb angle in navigation group was significantly lower than that in free-hand group at 1 week, 3 months after operation and at the last follow-up (all P<0.01). The VAS in navigation group and free-hand group was (3.2±0.7)points and (4.1±0.7)points at 1 week after operation, (2.4±0.6)points and (3.0±0.8)points at 3 months after operation, and (1.8±0.9)points and (2.6±0.7)points at the last follow-up, decreased significantly from that before operation [(8.4±0.8)points, (8.3±0.9)points] (all P<0.01). The VAS in navigation group was significantly lower than that in free-hand group at 1 week, 3 months after operation and at the last follow-up (all P<0.01). The ASIA score showed no significant difference within and between the two groups before operation, at 1 week, 3 months after operation and at the last follow-up (all P>0.05). Postoperative incision infection occurred in 1 patient in both groups ( P>0.05). Implant failure such as loosening or displacement was not observed in navigation group, and only occurred in 2 patients in free-hand group ( P>0.05). Conclusion:Compared with free-hand pedicle screw placement, O-arm assisted pedicle screw placement in the treatment of AO type C thoracolumbar fracture has advantages of rapid and accurate screw placement, good reduction and notable pain relief.