Objective To evaluate the therapeutic effect of Xiao'er Zhixie Paste (XZP) by using the young rat model of chronic diarrhea,and to explore its mechanism.Methods Chronic diarrhea model in young rats was induced by ig senna.Rats were ig with Montmorillonite powder of 1.62 g/kg,XZP of low,medium,and high dose (2.03,4.05,and 8.10 g/kg) for treatment.Loose stools rate,loose stool grade and diarrhea index were determined 1 and 3 d after treatment respectively.The water content of small intestine was measured and blood was collected for testing serum succinate dehydrogenase (SDH),amylase,D-xylose by colorimetric determination,testing serum D-lactic acid,IL-1 β,and TNF-α by Elisa after administration.Results The rate of loose stools in XZP 4.05 and 8.10 g/kg dose group,and diarrhea index in 8.10 g/kg dose group significantly reduced after the first treatment.The loose stools rate of XZP 2.03,4.05,and 8.10 g/kg dose group,diarrhea index,serum D-lactic acid level in 4.05,8.10 g/kg group significantly reduced,and serum D-xylose level in 8.10 g/kg dose group significantly increased 3 d after treatment.However,XZP had no significant effect on SDH,amylase activity and IL-1β,TNF-α levels.Conclusion XZP has obvious therapeutic effect on chronic diarrhea in young rats,the mechanism is to increase improve the absorptive function and permeability of intestinal tract.

OBJECTIVETo investigate the effects of immunologic effector cells to enhance apoptosis induced by adriamycin (ADR) in multi-drug resistant human breast cancer cell line MCF7/ADR.

METHODSThe immunologic effector cells were induced and expanded by IFN-gamma, McAb CD3, IL-1 and IL-2. The expression of P-glycoprotein (P-gp) and its relation to apoptosis in target cells were detected by TUNEL technique and immunohistochemical staining. Flow cytometry (FCM) was carried out to determine the expression level of human breast cancer related P185 antigen and the positive rate of Annexin V-FITC/PI expression. The subcellular distribution of ADR and Annexin V expression in the target cells were detected by fluorescence microscopy.

RESULTSThe immunologic effector cells down-regulated the expression of P185 and P-gp in MCF7/ADR cells. The accumulation and subcellular distribution of ADR in MCF7/ADR cells were increased after co-culture with the immunologic effector cells. After treatment with the immunologic effector cells in combination with ADR, apoptosis rate of the target cells was 10 times higher than that induced by ADR alone, and 13 times higher than that induced by the immunologic effector cells alone.

CONCLUSIONImmunologic effector cells can simultaneously down-regulate the expression of P185 and P-gp in MCF7/ADR cell line, and increase the apoptosis rate of MCF7/ADR cells in combination with ADR.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Breast Neoplasms ; immunology ; metabolism ; pathology ; Cell Line, Tumor ; Down-Regulation ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Female ; Humans ; Killer Cells, Lymphokine-Activated ; immunology ; Receptor, ErbB-2 ; metabolism

In order to investigate whether cytokine-induced killer (CIK) cells can induce apoptosis of bcr-abl(+) K562 cells, apoptosis of K562 cells and CEM cells induced by CIK cells, etoposide or camptothecin was detected with flow cytometry DNA assay. RT-PCR showed that K562 cells expressed the bcr-abl fusion gene, K 562 cells, K562 cells/etoposide or K562 cells/camptothecin groups showed no sub-G(1) peak. K562 cells/CIK cells group showed sub-G(1) peak (38.1%). CEM cells showed no sub-G(1) peak. CEM cells/camptothecin or CEM cells/etoposide groups showed sub-G(1) peak (23.5% or 32.3% respectively). CEM cells/CIK cells group showed sub-G(1) peak (45.4%). Etoposide or camptothecin did not induce apoptosis of K562 cells. CIK cells induce apoptosis of K562 cells. Bcr-abl fusion gene prevented apoptosis induced by etoposide or camptothecin, but did not prevent apoptosis induced by CIK cells. This property may support the observed adoptive immunologic effect of allogeneic bone marrow transplantation and donor lymphocyte transfusions of CML case relapsing after allogeneic bone marrow transplantation.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; immunology ; Camptothecin ; pharmacology ; Coculture Techniques ; Cytotoxicity, Immunologic ; Etoposide ; pharmacology ; Flow Cytometry ; Fusion Proteins, bcr-abl ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; K562 Cells ; drug effects ; immunology ; metabolism ; Killer Cells, Lymphokine-Activated ; cytology ; immunology

This study was purposed to establish a real-time fluorescent quantitative PCR (FQ-PCR) for quantifying SALL4 mRNA and to investigate its expression in different types of leukemia patients. SALL4 mRNA expression were measured in 60 leukemia patients of different periods and 10 normal controls sequentially by FQ-PCR. The results showed that the expression of SALL4 mRNA in de novo leukemia patients and relapsed patients was higher than that in controls (P < 0.05), which was significantly decreased at complete remission (CR). In relapsed patients, the expression of SALL4 mRNA increased slightly higher than that in de novo leukemia group, but the difference was not statistically significant (P > 0.05). However, the expression of SALL4 mRNA was low in CLL, T-ALL and AML-M3. The expression pattern of BMI-1 was same as SALL4, and the expression of BMI-1 positively correlated with that of SALL4 in leukemia (r = 0.825, P < 0.01). It is concluded that the detection of SALL4 gene expression in acute and chronic leukemia by real-time gTR-PCR displays high sensitivity and specificity. SALL4 gene may be one of indicators for monitoring the therapeutic outcome of partial leukemia and minimal residual disease.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Leukemia ; genetics ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; RNA, Messenger ; genetics ; Real-Time Polymerase Chain Reaction ; Transcription Factors ; genetics ; Young Adult

OBJECTIVETo analyze the improper pattern in mandarin monosyllable recognition test among the patients with Auditory Neuropathy (AN) in order to work out the common characteristics in speech recognition which might be suitable for diagnosis of AN.

METHODSSixteen AN patients (32 ears) were studied and 22 patients (32 ears) with sensorineural hearing loss (SNHL) were set for control. In accordance with audiogram pattern, all subjects were then divided into the up-type hearing (15 ears) and non up-type hearing (17 ears) groups. All 64 ears were tested in high intensity by mandarin monosyllable test material which we have developed before. Monosyllable performance scores from testing ears and improper patterns were recorded respectively. Eight improper patterns were then defined as follows: consonant only, vowel only, tone only, consonant and vowel, consonant and tone, vowel and tone, all phonemes and no response.

RESULTSThe score of patients with AN was lower than those patients with SNHL in monosyllable recognition test (P < 0.001). No significant difference was found between subgroup of up-type hearing loss and SNHL group in percentage correct scores of monosyllables, consonants, vowels, and tones statistically (P > 0.05), but significant lower score was found in subgroup of non up-type hearing loss compared with SNHL group in these 4 percentage scores concerned (P < 0.001). Chi square test presented a significant difference in improper pattern proportion between AN and SNHL groups (P < 0.001), which could be related to more proportional tone recognition in the former's incorrect items. Improper pattern proportions between two AN subgroups presented a significant difference statistically (P < 0.001), which could be related to a larger proportional recognition of tones and vowels in subgroup of up-type hearing loss compared with subgroup of non up-type hearing loss.

CONCLUSIONSA poor performance might be a major clinical feature identified AN from SNHL in mandarin tone recognition. There are significant differences between AN patients with up-type hearing loss and patients with non up-type hearing loss in performance of monosyllable recognition and improper pattern proportion of tones and vowels. A psychophysical testing may be a key potential in diagnosis of AN in further clinical application.

Adolescent ; Adult ; Audiometry, Speech ; Child ; Female ; Hearing Loss, Central ; physiopathology ; Hearing Loss, Sensorineural ; physiopathology ; Humans ; Language ; Male ; Middle Aged ; Speech Perception ; Young Adult

Objective: To observe the effects of functional training combined with self-made hand flexing training band in treatment of scar contracture after burn injury of dorsal hand. Methods: Forty-six patients with scar contracture after deep partial-thickness or full-thickness burn injury of dorsal hand hospitalized in our department from March 2013 to February 2015 were divided into routine training group (RT, n=18) and comprehensive training group (CT, n=28) according to their willingness. Two weeks after the wounds were healed, patients in group RT were treated with functional training of hands and self-made pressure gloves, while patients in group CT were treated with self-made hand flexing training band (consisting of nylon strap, flexing band, and velcro) on the basis of those in group RT. All patients were treated for 3 months. Before and after treatment, scar condition of affected hands was assessed with Vancouver Scar Scale (VSS). The range of motion of joints of affected hands was measured by Total Active Movement (TAM) Scale. The function of affected hands was evaluated by Carroll Upper Extremity Function Test. Data were processed with t test, chi-square test, and Mann-Whitney U test. Results: (1) The score of VSS in patients of group RT was (10.0±1.9) points before treatment and (4.4±1.4) points after treatment, with the improved score of (5.6±1.0) points. The score of VSS in patients of group CT was (10.5±1.8) points before treatment and (4.6±1.4) points after treatment, with the improved score of (5.9±1.2) points. There was no statistically significant difference in the improved score of patients between the two groups (t=0.834, P>0.05). The score of VSS in patients of groups RT and CT after treatment was significantly lower than that before treatment (with t values respectively 14.014 and 10.003, P values below 0.01). (2) Before treatment, the ratios of excellent and good results according to TAM were 2/9 in patients of group RT and 3/14 in group CT, with no statistical differences between them (χ²=2.140, P>0.05). After treatment, the ratio of excellent and good results according to TAM in patients of group CT (6/7) was higher than that in group RT (5/9, χ²=0.023, P=0.038). The ratios of excellent and good results according to TAM in patients of groups RT and CT after treatment were significantly higher than those before treatment (with Z values respectively -2.023 and -4.780, P values below 0.05). (3) The improved score of hand function in patients of group CT was (26±12) points, which was higher than (15±7) points in group RT (t=3.278, P=0.002). The score of hand function in patients of groups RT and CT after treatment was significantly higher than that before treatment (with t values respectively 2.628 and 6.125, P values below 0.05). There were no significant differences in grades of hand function of patients between the two groups before treatment (Z=-0.286, P>0.05). After treatment, the grade of hand function in patients of group CT was higher than that in group RT(Z=-1.993, P=0.046). The grades of hand function in patients of groups RT and CT after treatment were significantly higher than those before treatment (with Z values respectively -2.717 and -4.998, P values below 0.01). Conclusions For patients with scar contracture after burn injury of dorsal hand, early functional training combined with hand flexing training band can improve the range of motion of hand joints and functional recovery of hand, and the result was better than functional training alone.

Objective To analyze the cause, prevention and treatment of complications related to endovascular embolotherapy for ruptured intracranial aneurysms. Methods Sixty-eight patients with ruptured intracranial aneurysms received early endovascular embolization. Intraoperatively, artery spasm was relieved by papaverine infusion through microcatheter or balloon dilatation; parent artery occlusion by coils was treated by anticoagulation or antiplatelet treatment; ruptured aneurysms were embolized using mixed heparin; arterial thrombosis after embolization was ameliorated by the micro-catheter infusion of r-tPA thrombolytic. Results In the 68 cases, 8 cases suffered from complications, accounting for 11.77%. Among them, 1 case of extensive spasm of middle cerebral artery developed cerebral infarction and mild hemiplegia; 2 cases of localized spasm were improved well without sequelae; 1 case in which partial coils entered middle cerebral artery achieved a satisfied therapeutic outcome without infarction; 1 case with cerebral infarction due to coil dropping and 1 case with aneurysm rupture during operation received secondary embolotherapy without adverse consequences; one week after operation, 1 case died from aneurysm re-rupture; 1 case had hemiplegia owing to massive cerebral infarction. Conclusions It will do much benefit to secure the success of the operation by displaying the location, shape and size of the aneurysms as well as the relationship with the parent artery. Moreover, the proper choice of coils and well mastering of operative skills can decrease the incidence of complications, and adequate and prompt treatment of intraoperative complications can improve the prognosis of the patients obviously.

OBJECTIVETo investigate the specific anti-breast cancer immune response induced by dendritic cells (DC) loaded with trastuzumab and apoptotic Her-2+ breast cancer cells.

METHODSDCs were generated from healthy peripheral blood mononuclear cells (PBMCs) in the presence of recombinant cytokines GM-CSF, IL-4 and TNF-alpha. Mature DCs were harvested after 7 days' co-culture of PBMCs and trastuzumab-treated apoptotic SKBr3 cells. The morphologic characteristics and ultrastructure of the DC were observed under the inverted phase-contrast microscope and transmission electron microscope (TEM), respectively. Flow cytometry (FCM) was used to check the expression of several DC specific markers: CD14, CD1a, CD64, CD80, CD83, CD86, HLA-ABC and HLA-DR. DC-cytokine induced killer (DC-CIK) cells were prepared by co-culture of DCs and peripheral blood lymphocytes in the presence of anti-CD3 antibodies and human IL-2 at an appropriate concentration. The number of antigen-specific T cells was analyzed by human interferon gamma enzyme linked immunospot (ELISPOT) assay. MTT assay was employed to assess the lysis of breast cancer cell line induced by DC-CIK cells.

RESULTS5 minutes after the adding of DCs to SKBr3 cells pretreated with trastuzumab, the apoptotic SKBr3 cells were found to be circled by DCs. 48 hours later, many membrane-wrapped organelles of the apoptotic target cells in the cytoplasm of DCs were found by TEM. The majority of the organelles were degraded. Fewer organelles from the apoptotic cells were found in DCs without Herceptin. More than 60% in every group of DCs expressed a high-affinity receptor for IgG (FcgammaRI or CD64). CD14 expression on the mature DCs were comparatively lower, and HLA-DR and HLA-ABC expressions were higher in the trastuzumab group. The expression of CD1a, CD80, CD83 and CD86 in trastuzumab group were higher than those in immature DCs group (P < 0.05). ELISPOT assay suggests that the spot number of antigen-specific T cells were higher in trastuzumab group than that in the antigen unloaded DCs group (P < 0.05). The lysis of SKBr3 cells induced by the SKBr3 antigen loaded DC-CIK cells were 1.7 times higher than that of CIK.

CONCLUSIONThe lysis of SKBr3 cells induced by DC-CIK was increased after that DCs were combined with trastuzumab to capture antigen from SKBr3 cells. These findings support further investigation into the use of combination immunotherapy of the humanized monoclonal antibody, DC vaccines and immunological effector cells.

Antibodies, Monoclonal ; pharmacology ; Antibodies, Monoclonal, Humanized ; Apoptosis ; Cell Line, Tumor ; Coculture Techniques ; Cytokine-Induced Killer Cells ; immunology ; Cytokines ; metabolism ; Cytotoxicity, Immunologic ; immunology ; Dendritic Cells ; cytology ; immunology ; metabolism ; ultrastructure ; Humans ; Receptor, ErbB-2 ; metabolism ; Receptors, IgG ; metabolism ; Trastuzumab

OBJECTIVETo explore the feasibility of semi-nested PCR technique for detection of immunoglobulin heavy chain (IgH) clonal rearrangement in bone marrow of B-cell lymphoma patient and to further evaluate its clinicopathological value.

METHODSGene clonal rearrangement of IgH was detected by semi-nested PCR using primers of FR2 & FR3A in 105 bone marrow samples of patients with B-cell lymphoma. The PCR detection results were compared with the cytomorphology of bone marrow aspiration biopsy. The correlation between PCR detection results and clinicopathological factors were evaluated.

RESULTSAmong 105 cases of B-cell lymphoma, bone marrow involvement was detected by PCR technique in 48 cases (45.7%), while only 22 cases (21.0%) were detected by bone marrow cytological analysis. There was a significant difference between two methods (P < 0.05), and the concordance rate was 71.4%. The incidence of bone marrow involvement at the time of initial diagnosis detected by PCR technique was 30.8% for diffuse large B cell lymphoma (DLBCL), 25.0% for follicular lymphoma (FL), and 100.0% for small lymphocytic lymphoma (SLL), respectively. Bone marrow involvement detected by PCR detection correlated with Ann Arbor stage. Rate of clonal IgH gene rearrangement by PCR in early B-cell lymphoma was lower than that in advanced stage B-cell lymphoma patients (P = 0.02). There was no statistically significant difference in efficacy between patients with positive and negative results detected by PCR (P > 0.05). But difference in complete response (CR) rate (23.3% and 46.3%) had significant difference (P = 0.019).

CONCLUSIONSemi-nested PCR analysis may be an effective method for detection of abnormalities in bone marrow in patients with B-cell lymphoma and is superior to cytomorphology. The positive rate in patients with advanced Ann Arbor stage is higher than that in patients with early Ann Arbor stage, and patients with PCR negative result have more chances to achieved CR after treatment.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Biopsy ; methods ; Bone Marrow ; pathology ; Female ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Leukemia, Lymphocytic, Chronic, B-Cell ; drug therapy ; genetics ; pathology ; Lymphoma, Follicular ; drug therapy ; genetics ; pathology ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Polymerase Chain Reaction ; methods ; Remission Induction