An in vitro drug sensitivity assay was developed in this hospital. Human cancer cells from biopsy or surgery which were treated with different anticancer drugs were used and the incorporation efficiency of 2H-thymidine into DNA of cancer cells was compared with the control group.The experimental results demonstrate that mito-mycin C and fluorouracil are the most active drugs for the treatment of gastric cancer. Vincristin and Sarcandra glaber are less effective. As far as breast cancer is concerned the activated cyclophosphamide and fluorouracil are most effective, while mitomycin C and mustine hydrochloride are less effective. Bu-fonis venenum was shown to be inactive in gastric cancer and breast cancer.The method is simple and it can be used for the predictability studies in cancer clinics.

Polysaccharide from Lao Shan Polystictus versicolor possesses a significant inhibitory effect on transplanted S180 in ICR mice, and the inhibition rate after intraperitoneal injection of 10 mg/kg of the drug was 60-70%. Oral administration in certain dosage could also inhibit tumor growth.Polysaccharide was given intraperitoneally one week before the transplantation of ascitic sarcoma S180 It showed that the polysaccharide could prevent the growth of ascitic S180 effectively and prolong the life span of the mice. Peritoneal injection of the polysaccharide could enhance the capacity of macrophages to release H2O2 and of the spleenic lymphocytes to take 3H-TdR. It has been demonstrated that polysaccharied from Lao Shan Polystictus versicolor was not cytotoxic, and its anticarcinogenic effect was host mediated.

Objective To establish a mouse model of acute liver failure induced by lipopolysaccharide /D-galac-tosamine ( LPS/D-GalN) .Methods The optimum dose of LPS/D-GalN was determined by i .p.injection of eight differ-ent doses of LPS and D-GalN into 40 female C57BL/6 mice and observation of their survival time .Then, 32 female C57BL/6 mice were i.p.injected with the optimal dose of LPS/D-GalN and sacrificed at 0, 1, 4, 8 hours after the injec-tion, 8 mice in each group.The control mice received saline injection .Hepatic changes were observed by pathology and se-rum ALT, IL-6, MCP-1 and TNF-αwere measured by biochemistry or flow cytometry .Results LPS (2.5 mg/kg) and D-GalN (0.3 g/kg) were determined as the optimal dose for the establishment of mouse model of acute liver injury .Com-pared with the control group , the hepatocellular damages were progressing in a positive correlation with the time course after LPS/D-GalN administration .The level of serum ALT was significantly increased after LPS/D-GalN administration ( P <0.001).The levels of inflammatory cytokines IL-6, MCP-1 and TNF-αwere increased and reached a peak at one hour after LPS/D-GalN administration and then decreased almost to that of the control group 8 hours later(P<0.001).Conclusions The mouse model of acute liver injury is successfully established by LPS /D-GalN administration , and provide an effective animal model for the study of pathogenic mechanisms of acute liver failure and evaluation of therapeutic drugs .

Objective To compare and analyze the clinical characteristics of acute central cervical spinal cord injury with only upper extremity involvement and with both upper and lower extremity involvement.Methods A retrospective case control study was made on clinical data of 76 patients with acute central cervical spinal cord injury hospitalized from January 2010 to December 2013.Nerve injury involved was only upper extremity in 39 patients (upper extremity group),but both upper and lower extremities in 37 patients (upper-and lower-extremity group).In upper extremity group,there were 35 males and four females,age was 21-80 years [(52.5 ± 13.4) years],injury resulted from traffic accidents in 24 patients,ground-level falls in eight,high-level falls in six and heavy-object hit in one,and level of injury was C3/4 in 16 patients,C4/5 in 14 and C5/6 in nine.In upper-and lower-extremity group,there were 30 males and seven females,age was 36-78 years [(59.6 ± 9.7) years],injury resulted from traffic accidents in 16 patients,ground-level falls in 11,high-level falls in seven and heavyobject hit in three,and level of injury was C3/4in nine patients,C4/sin 18 and C5/6in 10.Sagittal diameter of the cervical spinal canal,maximal canal compromise,maximal spinal cord compression,degenerating factors of the cervical spine and treatment protocols were determined.Upper extremity function was assessed with the American spinal injury association (ASIA) score.Results There were significant differences between upper extremity group and upper-and lower-extremity group in sagittal diameter of the cervical spinal canal [(7.5 ± 1.5)mm ∶ (6.8 ± 1.2)mm],maximal canal compromise [(28.9 ±9.6)％ ∶ (34.9 ± 10.6)％],ASIA score at admission[(31.6 ± 11.8)points ∶ (22.7± 11.3)points)] and ASIA score at last follow-up [(46.2 ± 4.2) points ∶ (40.2 ± 4.0) points] (P ＜ 0.05),while the maximal spinal cord compression in upper extremity group [(15.7 ± 11.9)％] had no significant difference from that in upper-and lowerextremity group [(17.0 ± 10.6) ％] (P ＞ 0.05).Lower prevalence of posterior osteophyte of the vertebral body was noted in upper extremity group than upper-and lower-extremity group (15％ ∶ 51％) (P ＜0.01).Twenty patients (49％) in upper extremity group were surgically treated,while 31 patients (84％) in upperand lower-extremity group (P ＜ 0.05).Conclusions Compared to acute central cervical spinal cord injury with both upper and lower extremity involvement,the injury with only upper extremity involvement is much common in younger patients and is characterized by lowered frequency of osteophyte,large buffer space,mild nerve damage,preferred non-operation treatment and good prognosis.

Objective To obtain Chinese hamster ovary (CHO) cell lines that stably express a targeting complement inhibitor CR2-CD59.Methods The recombinant plasmid PEE14.1-CR2-CD59 was constru-cted by cloning the DNA fragment CR2-CD59 into plasmid PEE14.1,and the obtained plasmid was transfected into CHO cells by FuGENE 6.The clones with stable high expression of target fragment were selected by methionine sulfoximine (MSX),the expression of CR2-CD59 was analyzed by ELISA,SDS-PAGE and Western blotting analysis.Results Several stable expression clones were obtained,and CR2-CD59 was highly expressed in the secret form in CHO cells.SDS-PAGE analysis showed that the molecular weight of the recombined protein CR2-CD59 was consistent with the predicted one.ELISA and Western blotting results revealed that the CR2-CD59 could react with both anti-human CR2 and anti-human CD59 polyclonal antibodies.Compared with serum-containing medium,the protein was highly expressed in serum-free medium (P

Objective To express a recombinant protein TFPR1 ( the functional region of the snake venom proteins from Trimeresurus flavoviridis) in Pichia pastoris expression system. Methods The target gene was codon-optimized and synthesized according to the sequence of the conserved structural do-main of triflin and then cloned into the Pichia pastoris expression vector pPICZαA to construct the recombi-nant expression plasmid pPICZαA-TFPR1. The recombinant plasmid pPICZαA-TFPR1 was electroporated into the yeast strain X33. The transformed strains carrying expression plasmid were screened out with Zeocin and then induced by methanol to express the recombinant protein TFPR1. ELISA was performed for the screening of positive clones. SDS-PAGE and Western blot were used for further identification of the ex-pressed products. Results The recombinant plasmid pPICZαA-TFPR1 was successfully constructed. The recombinant protein TFPR1 was expressed in a secreted form at a molecular weight of 16×103. Conclusion The recombinant protein TFPR1 was successfully expressed in Pichia pastoris expression system, which laid a foundation for further researches on its biological function and application as an adjuvant.

Objective To analyze and compare the pathological changes of lung tissue in mice infected with the novel H7N9 influenza virus and 2009 pandemic H1N1 influenza virus, respectively, and to preliminarily study the mecha-nisms of acute lung injury induced by those virus infection .Methods SPF 6-week old BALB/c mice ( body weight 18-20 g, male∶female=1∶1) (n=3 in each subgroup) were intranasally infected with H7N9 virus and H1N1 virus, respec-tively.The behavior and survival time of mice after virus infection were observed and the survival rates were analyzed .The heart, liver, spleen, lung, kidney, intestines, and brain were collected at indicated time points for histopathological exami-nation using H&E staining .The distribution of virus antigen was detected by immunohistochemistry .The neutrophil infiltra-tion was also observed .The correlation of lung injury with virus replication and host immune responses was analyzed .Re-sults The lung and spleen injury of mice infected with H 7N9 virus was slighter and their survival rate (100%) was high-er than those of mice infected with H1N1 virus.The damages of the lung and spleen in H1N1virus-infected mice were more severe than that in H7N9 virus-infected mice, and all the 10 mice in this group died within 9 days after virus inoculation . The distributions of both the virus antigens were mainly in the bronchial epithelial cells , a few stromal cells and alveolar ep-ithelial cells .The levels of virus replication in the two groups were not significantly different .There were more intense neu-trophil infiltration in the lung and inflammatory response in the H 1N1 virus-infected mice than those in the H7N9 virus-in-fected mice .Conclusions There are some differences of the pathological characteristics and extent of lung injury in the mice infected with H7N9 virus and H1N1 virus, respectively.The virus replication is a precipitating factor but not the deci-sive factor of the lung injury , and there is a close relationship between the host immune responses and acute lung injury .

Objective To investigate prospectively the changes of quantitative parameters in dynamic contrast-enhanced MRI (DCE-MRI) in cervical cancer patients before and after neoadjuvant chemotherapy (NACT).Methods Thirty-eight patients with locally advanced cervical cancer (in stage Ⅰ B2,Ⅱ A2,Ⅱ B) underwent DCE-MRI one week before and 1 month after NACT.The patients were classified into two groups:significant reaction(sCR) group and non-sCR group.The DCE-MRI pharmacokinetics parameters (mean Ktrans,mean Kep,mean Ve and mean Vp) were measured and compared between the sCR and non-sCR groups.Receiver operating characteristic (ROC) curves were constructed to describe the diagnostic accuracy of the significant parameters and their decision thresholds.Results There were 22 and 16 patients in sCR and non-sCR groups,respectively.Before NACT,the mean Ktrans was higher (P＜0.05) but the mean Ve (P＜0.05) was lower in sCR group than those in non-sCR group,and these differences were statistically significant,respectively.After NACT,the mean Ktrans (P＜0.05) and the changed value of Ktrans (P＜0.05) were significantly lower in the sCR group compared with those in the non-sCR group.The remained parameters such as Vp and Kep had no statistically difference between the two groups.When combined the parameters values before and after treatment by using ROC curves,the area under curve (AUC) of pre-mean Ktrans and /Ktrans were 0.801 (P＜0.05),0.955 (P＜0.001).The optimal cut off value for distinguishing sCR from non-sCR were the pretreatment Ktrans (0.702 0) and/Ktrans (0.043 7),and their sensitivity and specificity were 77.3％,81.2 ％,95.5％ and 81.2％,respectively.Conclusion Quantitative parameters of DCE-MRI provided a new noninvasive way to reflect the changes of hemodynamics in cervical cancer patients with NACT.The quantitative parameters,such as pre-mean Ktrans and /Ktrans could predict the treatment efficacy more precisely.

Objective To compare the adjuvant activity of oil-in-water emulsion MF59 and aluminum hydroxide on immune responses to HIV-1 multi-epitope protein MEP1 in BALB/c mice. Methods HIV-1 multi-epitope protein MEP1 with MF59 or aluminum was prepared to intramuscularly vaccinate female BALB/c mice for three times. Serum and splenocytes were isolated from the mice 10 d after the first and last vaccination. Specific anti-MEP1 antibodies and the subclasses in serum were detected by ELISA. The num-bers of splenic lymphocytes secreting IFN-γ and IL-4 were measured by ELISPOT. Differences in humoral and cellular immune responses induced by the two different adjuvants were compared. Results After a sin-gle-dose immunization, aluminum adjuvant promoted early humoral and cellular immune responses to MEP1 compared with MF59. After the three-dose immunization, both aluminum adjuvant and MF59 promoted MEP1 to induce Th2-biased humoral immune response, while MF59 also enhanced a balanced Th1/Th2 cel-lular immune response. Conclusions Aluminum adjuvant was a more suitable adjuvant than MF59 for HIV-1 multi-epitope protein MEP1.

Objective:To investigate the clinical safety and efficacy of tirofiban in the treatment of hemiplegic stroke warning syndrome.Methods:Patients with hemiplegic stroke warning syndrome admitted to Jining First People's Hospital without receiving intravenous thrombolysis from January 2018 to May 2020 were enrolled retrospectively. Some patients were given tirofiban intravenous infusion for at least 24 h in acute phase, then received oral antiplatelet therapy (tirofiban group); some only received aspirin+ clopidogrel dual antiplatelet therapy (control group). The primary endpoint was muscle strength at the paralytic side and National Institutes of Health Stroke Scale (NIHSS) score at day 7 after onset. The secondary endpoint was the modified Rankin Scale (mRS) score at 3 months after onset, and ≤2 was defined as good clinical outcome. The safety endpoint was the bleeding events during treatment. Multivariate logistic regression analysis was used to determine the independent influencing factors of clinical outcome. Results:A total of 30 patients with hemiplegic stroke warning syndrome were enrolled, including 19 (63.3%) in the tirofiban group and 11 (36.7%) in the control group. There was no significant difference in baseline clinical data between the two groups, and no drug-related bleeding complications occurred during treatment. The muscle strength at paralytic side and NIHSS score at day 7 after onset, NIHSS score at discharge and good clinical outcome rate at 3 months in the tirofiban group were significantly better than those in the control group, and the differences were statistically significant (all P<0.05). Multivariate logistic regression analysis showed that tirofiban was an independent protective factor for good outcome after adjusting the NIHSS score at the beginning of treatment (odds ratio 0.040, 95% confidence interval 0.040-0.449; P=0.009). Conclusions:Tirofiban is safe and effective in the treatment of patients with hemiplegic stroke warning syndrome in acute phase. It can effectively block the progress of the disease, improve the outcome of patients, and will not increase the risk of bleeding.