The ScFv gene containing Nco I and Bam HI was amplified by PCR, and inserted into the prokaryotic expression vector pHOG21 with Nco I and Bam HI digestion. After screening, high-level expression recombinant XL1-Blue (pHOG2E3) were obtained, and the different expression levels of ScFv were detected by SDS-PAGE and EUSA. When the strain was induced by 0.5 mmol/LIPTG and 0.4mol/L sucrose in the culture medium LB at 37℃ for 6 hours, the high level production of ScFv protein was obtained. The molecular weight of the expressed ScFv is 31, 000 D. The ScFv was mainly in the form of inclusion body, but it could also be in the culture medium and soluble periplasmic content. Above all, the ScFv protein could neutralize the phospholipase C activities of alpha-toxin of Gostridium perfrigens type A.

Objective To study the regulatory T cells (Treg-cell) frequencies in patients with childhood acute leukemia and evaluate its clinical application value by investigating the relationship between the increasing numbers of Treg cells and minimal residual disease of leukemia (MRD), Methods Foxp3-FITC/CD25-PE/CD4-PerCP/CD3-APC four-color staining flow cytometry was established to identify Treg cells. Treg cells frequencies both in 10 healthy controls and in 33 patients with newly diagnosed childhood acute leukemia ( B-ALL 17 cases, T-ALL 9 cases, AML 7 cases) were detected. The possibility of the accumulation of Treg cells being the prognostic marker for acute leukemia was evaluated by comparing the results of Treg cells frequency with that of MRD. Results The percentage of Treg cells in CD+4 CD+3 T cells was M = 8. 09% in normal bone marrows, which was significantly different from the results in the bone marrows of newly diagnosed childhood acute leukemia ( M = 12.77% , U = 3.41, P < 0.01 ), but it showed no significantly differences among B-ALL, T-ALL and AML groups. No association was observed between the expression of Treg cells and clinical-biologic characteristics studied. In addition, Treg cells frequency in MRD positive group was significantly different from that in MRD continuously negative group ( M = 14. 74% vs M=11.3%, t =252.5,P<0.05). Conclusions Consistent with results from solid tumor, the study identifies a significantly increased numbers of Treg cells in patients with childhood acute leukemia. The situation of accumulation of Treg cells is closely associated with MRD results during chnical remission. High level of Treg cells may cause poor prognosis and increase the possibility of relapse.

Objective To observe the effects of epidermal growth factor (EGF) on cell proliferation and cell cycle of cultured bovine corneal endothelial cells. Methods Bovine corneal endothelial cells were cultured with different concentrations of hEGF (1, 10 and 100 ng/mL). MTT test was performed to evaluate the cell proliferation. The bovine corneal endothelial cells were divided into two groups:control group (cultured in DMEM) and EGF-stimulated group (cultured in DMEM with EGF). Flow cytometry was performed to determine the cell cycle phases on the third and seventh day. Results Compared with the control, EGF enhanced the cell proliferation in a dose-related response. 10 ng/mL and 100 ng/mL EGF were much more effective than 1 ng/mL.On the third day, S phase cells accounted for 24.5% and G_2-M phase cells 0.08% in the control group,while 24.6% and 0.06%, respectively in the EGF-stimulated group. However, on the seventh day, those came to 20.8% and 0.41% in the control group, and 18.2% and 1.55% in the EGF-stimulated group,indicating a significant change in the cell cycle (P

Objective To evaluate the effect of losecplatin combined with compound matrine in the intraperitoneal perfusion treatment of ovarian cancer with malignant ascites and its influence on the levle of serum tumor necrosis factor alpha(TNF-) of ovarian cancer marker.Methods 60 ovarian cancer patients with malignant ascites were divided into three groups:losecplatin combined with compound matrine perfusion group(combination group),single losecplatin perfusion group(losecplatin group) and single compound matrine perfusion group(compound matrine group),20 cases in each group.Before treatment,all the three groups were drained intraperitoneal fluid,then given the above-mentioned group of intraperitoneal perfusion therapy.The effects,side effects and serum TNF- levels of the three groups were compared.Results The patients of the three groups were successfully completed treatment,the effective rate of the combination group was 95%,which of the losecplatin group was 60%,which of the compound matrine group was 55%.The effective rate of the combination group was significantly higher than that of the single drug group(χ2=7.025,P<0.05),and the adverse reaction of the combination group was not significantly increased(χ2=1.026,P>0.05).The serum TNF- levels of the three groups after the perfusion treatment were significantly decreased(t=15.40,13.82,8.90,all P<0.05),TNF- level of the combination group was significantly lower,the difference was statistically significant(F=9.719,P<0.05).Conclusion Losecplatin combined with compound matrine in the intraperitoneal perfusion is a more effective method for the treatment of ovarian cancer with malignant ascites,which is worthy of promotion.

OBJECTIVETo study the effect of Fuzheng Sanjie recipe in regulating tumor-associated macrophages (TAMs) in Lewis lung cancer mice.

METHODEfforts were made to establish the Lewis lung cancer mouse model, weigh tumors and calculate the anti-tumor rate. The immunohistochemical method was used to examine the infiltration degree of CD68 + in tumor tissues in each group. ELISA was used to examine the content of IFN-γ, TGF-β, IL-4, IL-13, IL-6, IL-10, IL-12, TNF-α in mice serum.

RESULTCompared with the tumor-bearing model group, all of the other groups showed higher tumor inhibition rates, i. e. 50.28% for the DDP group, 34.37% for the TCM-preventing group and 66.76% for the Chinese and western medicine group, with statistical difference (P < 0.05), but without statistical difference in the infiltration degree of CD68+. The expressions of the IFN-γ, IL-6, IL-12 in tumor-bearing groups were lower than that in the blank control group, but with higher contents of IL-4, IL-13, TGF-β. Intervened with different drugs, there were significant differences in content among some relevant cytokines (P < 0.05), as well as statistical differences among the TCM prevention group, the Chinese and western medicine group and the tumor-bearing control group (P <0. 05) , but without statistical difference in TNF-α and IL-10 content from the tumor-bearing control group (P < 0.05).

CONCLUSIONFuzheng Sanjie recipe could reverse the immune remodeling effect and control the tumor growth by down-regulating the expressions of IL-4, IL-13, TGF-α in lung cancer immune microenvironment and up-regulating the expression of IFN-γ.

Animals ; Cell Line, Tumor ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Interleukin-10 ; blood ; Interleukin-12 ; blood ; Interleukin-13 ; blood ; Lung Neoplasms ; blood ; drug therapy ; immunology ; Macrophages ; drug effects ; immunology ; Male ; Mice ; Mice, Inbred C57BL ; Transforming Growth Factor beta ; blood ; Tumor Necrosis Factor-alpha ; blood

The structural theory and fabrication of a self-designed rotating auxiliary spring in edgewise technique were described. Rotating auxiliary spring was applied to correct rotated tooth in clinic. It indicated that rotating auxiliary spring was an easy-fabricated, convenient-manipulated device, and could produce an effective treatment results in correcting rotated tooth, especially in the final adjustment stage.
Humans ; Malocclusion ; Tooth Movement Techniques

Objective To observe the clinical effect of intensity modulated radiation therapy (IMRT) combined with apapatinib mesylate in the treatment of elderly patients with locally advanced cardia adenocarcinoma and its effect on vascular endothelial growth factor receptor (VEGFR). Methods Forty-six elderly patients with locally advanced cardia cancer who were unwilling to accept surgery or couldn't get complete removal of cancers in Xuzhou Hospital Affiliated to Jiangsu University between January 2015 and April 2016 were collected. All the patients were randomly divided into the control group (23 cases) and the observation group (23 cases) according to the random number table method. The control group received radiotherapy alone. In the observation group, oral apatinib (500 mg/d) was taken in the first day of radiotherapy after breakfast until the disease progress or death occurred. Results The total effective rate was 92 % (19/23) in the observation group and 60 % (10/23) in the control group. There was a significant difference between the two groups(χ2=5.86,P <0.05). After treatment, the average level of VEGFR in both groups was decreased[(76.3±4.9)vs.(55.0±2.3)pg/ml],and there was a significant difference between the two groups (t = 3.93, P < 0.05). The common adverse reactions were blood adverse reactions and gastrointestinal reactions. The incidence rate of gastrointestinal reaction was both 83 %, and the blood adverse reaction was 100 %, and there was no significant difference between the two groups (both P > 0.05). Hypertension, proteinuria and rash response in the observation group were increased compared with the control group,but noⅣ grade of adverse reactions occurred. The median progression-free survival time was 10 months in the observation group and 8 months in the control group respectively (P = 0.01). Conclusion IMRT combined with apatinib in the treatment of elderly patients with locally advanced gastric cardia has a favorable efficacy and tolerance.

OBJECTIVETo elucidate the clinical features of patients at early stage of severe acute respiratory syndrome (SARS).

METHODSFifty-three cases of early SARS were studied retrospectively. The data reviewed included those of epidemiology, clinical manifestations, laboratory investigation and roentgenology.

RESULTSThe patients consisted of 24 men and 29 women, aged 10.85 years (mean 38+/-16.7 years), including 9 infected health-care professionals (17.0%). The mean incubation period was 7.3+/-7.0 days (3.14 days). The onset symptoms were characterized by fever (100%), cough (49.1%), maylgia (24.5%), shortness of breath (20.8%), malaise (17.0%),and diarrhea (5.7%). Routine blood test during the first to the fifth day of the disease revealed WBCs less than 4.0x10(9) /L in 33 patients (62.3%), 4.0-10.0x10(9)/L in 18 patients (34.0%), lymphopenia in 36 patients (67.9%), and PLT less than 100.0x10(9) in 7 patients (13.2%). The main abnormal X-ray finding was single (75.4%) or bilateral (15.1%) localized patchy clouding opacity. The decrease of arterial partial pressure of oxygen occurred in 26 patients (49.1%). The damage of several organs was common, including the elevated ALT or AST in 20 patients (37.7%), elevated BUN or SCR in 6 patients (11.3%) and elevated LDH or CK or HBDH in 23 patients (43.4%).

CONCLUSIONThe clinical manifestations of SARS at the early stage were complex. The close monitoring of the blood cell counts, the blood gas analysis and chest radiography might be crucial to the early diagnosis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blood Cell Count ; Blood Gas Analysis ; Child ; Early Diagnosis ; Female ; Humans ; Male ; Middle Aged ; Radiography, Thoracic ; Retrospective Studies ; Severe Acute Respiratory Syndrome ; diagnosis ; epidemiology

OBJECTIVEThe aim of this study was to detect the pre- and post-treatment serum carcinoembryonic antigen (CEA) levels after 4 weeks of EGFR-TKIs treatment in advanced non-small cell lung cancer (NSCLC) patients to evaluate the clinical value of CEA in the prediction of chemotherapy response and prognosis in those patients.

METHODSPre- and post-treatment serum CEA levels of the patients were measured with immunoradiometric kits after 4 weeks of EGFR-TKIs treatment to evaluate the relationship between chemotherapy response and prognosis.

RESULTSAfter 4 weeks of EGFR-TKIs treatment, one patient in the total of 75 patients (1.3%) achieved complete response (CR), 17 patients (22.7%) achieved partial response (PR), 31 patients (41.3%) achieved disease stable (SD) and 26 patients had progressive disease (PD). The radiological objective response rate(ORR) and disease control rate (DCR) were 24.0% and 65.3%, respectively. The median survival time (MST) of all patients was 8.1 months. The MST of SD patients was similar to that in the OR patients (P = 0.06), but both longer than that in the PD patients (P < 0.001). The MST of DC patients was similar to that in OR patients (P = 0.358), but longer than that in PD patients (P < 0.001). Serum CEA levels decreased ≥ 32% and ≥ 61% were closely related with the objective response and disease control. The median survival time (MST) of patients with serum CEA decreased ≥ 32% was longer than those with CEA decreased < 32% (9.5 months vs 6.7 months, P < 0.0001). The MST of patients with serum CEA decreased ≥ 32% was similar to those with CEA decreased ≥ 61% (9.5 months vs 10.5 months, P = 0.370), but both longer than those with CEA decreased < 32% (6.7 months, P < 0.001). Cox multivariate survival analysis confirmed that serum CEA level decreased ≥ 32%, CEA level decreased ≥ 61%, PS score, and DC are independent prognostic factor, but not OR.

CONCLUSIONSTo advanced NSCLC patients, the disease control rate (DCR) may be more suitable than objective response rate (ORR) as an indicator in predicting the efficacy and prognosis in advanced NSCLC patients. Serum CEA levels decreased ≥ 32% may be a reliable indicator to determine the therapeutic efficacy of EGFR-TKIs.

Aged ; Antineoplastic Agents ; therapeutic use ; Carcinoembryonic Antigen ; blood ; Carcinoma, Non-Small-Cell Lung ; blood ; drug therapy ; pathology ; Erlotinib Hydrochloride ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; blood ; drug therapy ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Proportional Hazards Models ; Protein Kinase Inhibitors ; therapeutic use ; Quinazolines ; therapeutic use ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; Remission Induction ; Survival Rate

Na⁺/H⁺ exchangers (NHEs) have been shown to be involved in regulating cell volume and maintaining fluid and electrolyte homeostasis. Pooled evidences have suggested that loss of Na⁺/H⁺ exchanger isoform 8 (NHE8) impairs intestinal mucosa. Whether NHE8 participates in the pathology of infectious colitis is still unknown. Our previous study demonstrated that somatostatin (SST) could stimulate the expression of intestinal NHE8 so as to facilitate Na⁺ absorption under normal condition. This study further explored whether NHE8 participates in the pathological processes of infectious colitis and the effects of SST on intestinal NHE8 expression in the setting of infectious colitis. Our data showed that NHE8 expression was reduced in Citrobacter rodentium (CR) infected mice. Up-regulation of NHE8 improved diarrhea symptom and mucosal damage induced by CR. In vitro, a similar observation was also seen in Enteropathogenic E. coli (EPEC) infected Caco-2 cells. Seglitide, a SST receptor (SSTR) 2 agonist, partly reversed the inhibiting action of EPEC on NHE8 expression, but SSTR5 agonist (L-817,818) had no effect on the expression of NHE8. Moreover, SST blocked the phosphorylation of p38 in EPEC-infected Caco-2 cells. Taken together, these results suggest that enhancement of intestinal NHE8 expression by SST could ameliorate the symptoms of mice with infectious colitis.
Absorption ; Animals ; Anti-Inflammatory Agents ; Caco-2 Cells ; Cell Size ; Citrobacter rodentium ; Colitis* ; Diarrhea* ; Enteropathogenic Escherichia coli ; Homeostasis ; Humans ; In Vitro Techniques ; Intestinal Mucosa ; Mice* ; Pathologic Processes ; Pathology ; Phosphorylation ; Somatostatin* ; Up-Regulation*