Objective To investigate the role of Ca2+/calmodulin-dependent protein kinase Ⅱ on pain behavior in a mouse model of bone cancer pain. Methods 40 male C3H/HeN mice were divided randomly into 5 groups:sham group (S group, n=8) ,control group (C group, n=8) and KN93 treat group (T1, n=8;T2, n=8;T3, n = 8 ). Group C and T were induced mouse models of bone cancer pain by intra-left-femur inoculations of osteolytic NCTC2472 cells while group S were injected only α-MEM. On the 14 d after inoculations,group S and C received intrathecal injection of 20% DMSO 5 μl . While group T1, T2, T3 received intrathecal injection of KN93 15nmol,30nmol,60nmol which dissolved in 5 μl 20% DMSO respectively. Mice received pain behavior tests including quantification of spontaneous flinches, paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) before and at 0.5 h,2 h,4 h,8 h after administration. Results Treatment with KN93(15 nmol) have no effect on bone cancer pain,while treatment with KN93(30 nmol,60 nmol) can dose-dependently reverse quantification of spontaneous flinches, mechanical allodynia and thermal hyperalgesia which were induced by bone cancer pain, At 0. 5h after administration, the quantification of spontaneous flinches of the two groups ( (7.25 + 1.49 ), (4. 12 + 1.36 ) ) were decreased when compared with control group ( 11.62 + 1.92 ),PWMT((1.28 +0.14)g;(1.75 +0.46)g),PWTL((14.64 +2.12) s; (16.85 + 1.61)s)were increased when compared with control group ( (0.47 + 0. 16) g, ( 11.32 + 1.68 ) s) (P ＜ 0.05 ＝. The effect lasts for at least 4 h and disappears at 8 h. Conclusion CaMK Ⅱ may play an important role in the mechanism of bone cancer pain. Intrathecal KN93 injection can effectively attenuated bone cancer pain.

ObjectiveTo investigate the effect of KN93,a CaMKII inhibitor,on the spinal NR2B expression in the bone cancer pain mouse and its underlying mechanism.MethodsThirty-six male C3IL/IIeJ mice were randomly divided into 3 groups:sham group( S,n =8 ),bone cancer pain group( BP,n =8 ) and KN93 group ( K,n=20).The mouse model of bone cancer pain was established by intra-femur inoculation of osteolytie NCTC 2472 cells in BP and K groups.At 14d post operation,mice in K group received intrathecal injection of 60nmol KN93/5μl in 20％ DMSO and mice in BP group and S group received 20％ DMSD 5μl respectively.Eight mice were selected randomly from each group at (1)d before inoculation,at 1 h before administration and at 1,2,4,24h after administration( T0-5 ) to be measured the paw withdrawal threshold(PWT) stimulated by von Frey filaments.Another 3 mice were sacrificed at the corresponding time point and the spinal cord L3 -5 were obtained for determination of NR2B expression by western blot.ResultsPWT was significantly decreased in group BP( (0.50 ± 0.11 ) g) and K( (0.52 ±0.10)g),except for group K at T3(P＞0.05),and NR2B cxpression up-regulated at T2-5 in BP( 1.78± 0.34),K groups ( ( 1.11 ± 0.14),(0.73 ± 0.03 ),( 1.11 ± 0.15 ),( 1.89 ± 0.32 ) ) compared with S group ( ( 1.78 ± 0.31 ) g,(0.33 ± 0.04),P ＜ 0.05 ).Compared with group BP,PWT was increased and NR2B expression down-regulated at T2-4 in group K.In contrast to T1,PWT at T2-4 upgraded in group K(P＜0.05 ),but no significant difference was observed in other groups (P＞ 0.05 ).ConclusionIntrathecal injection of KN93 can attenuate bone cancer pain in mice through inhibiting NR2B with a time-dependent manner and spinal CaMKII-NR2B pathway may participate in the development of bone cancer pain.

Objective To study the therapeutic effects of Ag85A plasmid DNA vaccines in a mouse model of multi-drug resistant-(MDR-) Mycobacterium tuberculosis infection. Methods BALB/c mice were infected with Mycobacterium tuberculosis clinical strain HB361 with isoniazid and rifampin resist-ance by intratail-vein injection and were subsequently divided into 6 groups. At the third day after infection, the mice were treated with saline (group A), vector pVAX1 (greup B), rifampin (group C), vaccae (group D), Ag85A plasmid DNA vaccines (group E),rifampin and Ag85A plasmid DNA vaccines (group F) for 60 d. The lungs and spleens from the mice were taken and their pathological changes, weight and number of myeobacterial colony were examined at the third week after the end of treatment. Results At third week af-ter the end of treatment, the gross pathological observation and histopathological examination in lung showed that the lung lesions were limited, the profile of the alveoli was relatively clear, and normal structure could be seen in 2/3 areas of the lung sections in group D, E and F. The extent of lung lesion was 50% in group D,20% in group E and F. The pathological changes in group A, B, and C were more severer than those in group D, E and F. Compared with group A, the colony-forming units (CFU) in the lungs from mice in group D,E and F decreased 52%, 68%, 78%, respectively. The CFU in the spleens from mice in group D,E and F decreased 48%, 65%, 79%, respectively. Conclusion Ag85A plasmid DNA vaccines alone or Ag85A plasmid DNA vaccines along with chemotherapy have significant therapeutic effects on the mouse model of MDR-Mycobacterium tuberculosis infection.

Chromosomal region maintenance 1 (CRM1) is associated with an adverse prognosis in glioma. We previously reported that CRM1 inhibition suppressed glioma cell proliferation both in vitro and in vivo. In this study, we investigated the role of CRM1 in the migration and invasion of glioma cells. S109, a novel reversible selective inhibitor of CRM1, was used to treat Human glioma U87 and U251 cells. Cell migration and invasion were evaluated by wound-healing and transwell invasion assays. The results showed that S109 significantly inhibited the migration and invasion of U87 and U251 cells. However, mutation of Cys528 in CRM1 abolished the inhibitory activity of S109 in glioma cells. Furthermore, we found that S109 treatment decreased the expression level and activity of MMP2 and reduced the level of phosphorylated STAT3 but not total STAT3. Therefore, the inhibition of migration and invasion induced by S109 may be associated with the downregulation of MMP2 activity and expression, and inactivation of the STAT3 signaling pathway. These results support our previous conclusion that inhibition of CRM1 is an attractive strategy for the treatment of glioma.

Chromosomal region maintenance 1 (CRM1) is associated with an adverse prognosis in glioma. We previously reported that CRM1 inhibition suppressed glioma cell proliferation both in vitro and in vivo. In this study, we investigated the role of CRM1 in the migration and invasion of glioma cells. S109, a novel reversible selective inhibitor of CRM1, was used to treat Human glioma U87 and U251 cells. Cell migration and invasion were evaluated by wound-healing and transwell invasion assays. The results showed that S109 significantly inhibited the migration and invasion of U87 and U251 cells. However, mutation of Cys528 in CRM1 abolished the inhibitory activity of S109 in glioma cells. Furthermore, we found that S109 treatment decreased the expression level and activity of MMP2 and reduced the level of phosphorylated STAT3 but not total STAT3. Therefore, the inhibition of migration and invasion induced by S109 may be associated with the downregulation of MMP2 activity and expression, and inactivation of the STAT3 signaling pathway. These results support our previous conclusion that inhibition of CRM1 is an attractive strategy for the treatment of glioma.

Objective To observe the value of clinical and CT radiomics features for predicting microsatellite instability-high(MSI-H)status of gastric cancer.Methods Totally 150 gastric cancer patients including 30 cases of MSI-H positive and 120 cases of MSI-H negative were enrolled and divided into training set(n=105)or validation set(n=45)at the ratio of 7∶3.Based on abdominal vein phase enhanced CT images,lesions radiomics features were extracted and screened,and radiomics scores(Radscore)was calculated.Clinical data and Radscores were compared between MSI-H positive and negative patients in training set and validation set.Based on clinical factors and Radscores being significant different between MSI-H positive and negative ones,clinical model,CT radiomics model and clinical-CT radiomics combination model were constructed,and their predictive value for MSI-H status of gastric cancer were observed.Results Significant differences of tumor location and Radscore were found between MSI-H positive and negative patients in both training and validation sets(all P＜0.05).The area under the curve(AUC)of clinical model,CT radiomics model and combination model for evaluating MSI-H status of gastric cancer in training set was 0.760,0.799 and 0.864,respectively,of that in validation set was 0.735,0.812 and 0.849,respectively.AUC of clinical-CT radiomics combination model was greater than that of the other 2 single models(all P＜0.05).Conclusion Clinical-CT radiomics combination model based on tumor location and Radscore could effectively predict MSI-H status of gastric cancer.

Objective To evaluate the effect of body mass index (BMI) on early prognosis of the recipients after lung transplantation. Methods Clinical data of 307 lung transplant recipients were retrospectively analyzed. According to preoperative BMI, all recipients were divided into the low (BMI <18.5 kg/m², n=114), normal (18.5 kg/m²≤BMI <24 kg/m², n=145) and high (BMI≥24.0 kg/m², n=48) BMI groups, respectively. Baseline data including age, sex, blood type, BMI, preoperative complications, preoperative pulmonary hypertension, and intraoperative use of extracorporeal membrane oxygenation (ECMO) of all recipients were compared among three groups. The survival rate of all recipients was estimated by Kaplan-Meier curve and the survival curve was delineated. The differences of survival rate were analyzed by log-rank test. The 30-, 90- and 180-d mortality risk of all recipients after lung transplantation in different BMI groups was compared by multivariate Cox regression analysis. Results There were significant differences in age and sex of recipients among three groups (both P<0.05). There was a significant difference regarding the 180-d survival rate after lung transplantation among different BMI groups (P<0.05). Multivariate Cox regression analysis showed that the 90-d mortality risk after lung transplantation in the high BMI group was 2.295 times higher than that in the normal BMI group [hazard ratio (HR) 2.295, 95% confidence interval (CI) 1.064-4.947, P=0.034]. In the high BMI group, the 180-d mortality risk after lung transplantation was 2.783 times higher compared with that in the normal BMI group (HR 2.783, 95%CI 1.333-5.810, P=0.006), and the 180-d mortality risk in the low BMI group was 2.181 times higher than that in the normal BMI group (HR 2.181, 95%CI 1.124-4.232, P=0.021). Conclusions Compared with the recipients with normal BMI, their counterparts with high and low preoperative BMI have higher mortality risk early after lung transplantation. Adjusting preoperative BMI to normal range contributes to improving early prognosis of lung transplant recipients.

Blood-brain barrier (BBB) damage after ischemia significantly influences stroke outcome. Compound LFHP-1c was previously discovered with neuroprotective role in stroke model, but its mechanism of action on protection of BBB disruption after stroke remains unknown. Here, we show that LFHP-1c, as a direct PGAM5 inhibitor, prevented BBB disruption after transient middle cerebral artery occlusion (tMCAO) in rats. Mechanistically, LFHP-1c binding with endothelial PGAM5 not only inhibited the PGAM5 phosphatase activity, but also reduced the interaction of PGAM5 with NRF2, which facilitated nuclear translocation of NRF2 to prevent BBB disruption from ischemia. Furthermore, LFHP-1c administration by targeting PGAM5 shows a trend toward reduced infarct volume, brain edema and neurological deficits in nonhuman primate