Xiangxi (the western part of Hunan province) Liu's infantile tuina,as one main school of current infantile tuina in China,highlights the compatibility of the specific points of Wujing in children,the idea of treatment by syndrome differentiation,and produces unique efficacy in the treatment of common diseases in children.Exogenous fever in children can be treated with this method with excellent efficacy.Based on the clinical experience and effective cases treated by this tuina school,the authors elaborated the clinical thought and experience from the perspective view of tuina,for the promotion of Xiangxi Liu's infantile tuina in clinic.

AIM: To construct prokaryotic expression vector of His-tagged human IP-10 for further study of its biological function in the inflammatory response. METHODS: The coding sequence of IP-10 lacking signal peptide was amplified from human lung cDNA library by polymerase chain reaction (PCR) and the fragment was cloned into pET-14b plasmid for the construction of His-tagged fusion protein expressing vector, pET-14b/IP-10. After being identified by enzyme digestion and sequencing, the recombinant vector was transformed into a strain of E. coli, BL21 (DE_3). The expression of His-tagged fusion protein was induced with IPTG and purified with Ni+-NTA affinity chromatography. Then the chemotactic activity of IP-10 was determined by transwell migration assay on THP-1 cells. RESULTS: The construction of pET-14b/IP-10 recombinant vector was proved by enzyme digestion and sequencing. The fusion protein IP-10, which was purified by a routine Ni+ affinity method, had an activity on the induction of cell migration of THP-1. CONCLUSION: We successfully construct IP-10 fusion protein expressing vector and get the fusion protein with high bioactivity, which provides essential materials for the future studies on IP-10.

0.05)and the TNM staging (P=0.55).A mild elevated compared other pathological classification was found in small cell lung cancer (0.191?0.275).Conclusions The results showed that RFQ-PCR was suitable for measurement of the mRNA level of PLKI in bronchoscopic bioptic specimens.This study suggest elevated expression of PLK1 might play a important role in development of lung cancer,so that PLK1 might be a potential tumor marker for Lung cancers.Advanced studies will be needed to clarify that PLKI mRNA level do not relate to TNM staging and pathological classification.

Objective To observe the process of Vibrio vulnificus inducing dendritic cell strain apoptosis.Methods We established the mixed culture model of mouse dendritic cell ( DC 2.4 strain) and Vibrio vulnificus( Vv1.1758 strain ),analyzed morphological characteristics of cell apoptosis by DAPI fluorescence staining,detect DNA fragmentation level of apoptosis cells by DNA Ladder assay,analyze DC2.4 apoptosis rate by Annexin V FITC/PI staining,determine activities of caspase-3 and caspase-8 by means of spectrophotometric method and detect changes of mitochondrial transmembrane potential ( △ Ψm ) by JC-1 fluorescence labeling.Results After Vv1.1758 strain and DC2.4 cell were mixed and cultured for 4 h,DAPI fluorescence staining showed typical apoptosis characteristics-chromatin condensation and marginalization; DNA agarose gel electrophoresis showed apoptosis band; apoptosis rates at 2,4 and 6 h were respectively (37.8±9.8) ％,(54.3 ± 12.7 ) ％ and ( 68.2± 14.6 ) ％ ; Mitochondrial transmembrane potentials (△Ψm) at 1 h,2 h and 4 h reduced by 7.1％,16.1％ and 46.7％ respectively; caspase-8 activity increased at 1.5 h and reached the peak at 2 h [ (2.48±0.19) U/μg],while caspase-3 activity started to increase at 3h and reached the peak at 4 h [ ( 1.91 ±0.16) U/μg ].Conclusion Vibrio vulnificus could induce dendritic cells by two pathways: reducing mitochondrial transmembrane potential and activating caspase-8 promoter and finally activate effector caspase-3 to promote apoptosis.

Objective To analyze the incidence and risk factors for regional nodal failure(RNF) and chest wall recurrence(CWR) in T1 or T2 breast cancer patients(median age 44 year-range 26-72) with 1-3 positive axillary nodes treated with postmastectomy radiotherapy limited to the regional nodes.Methods From 1990 to 1999,320 patients were treated with postmastectomy(radical or modified radical) radiother- apy confined to the supraclavicular and internal mammary nodes with a median dose of 50 Gy in 25 fractions over 5 weeks.The median number of nodes examined was 8 (range 1-24).The median lymph nods rate (LNR) was 25% (range 5%-100%).Results The 5-year overall survival rate and disease free survival rate was 89.7% and 83.4%,respectively.The 5-year RNF and CWR was 7.9% and 5.7%,respectively. The 5-year RNF in patients with LNR＜30% and≥30% was 4.4% and 14.0% (P=0.002).The 5-year CWR in the subgroups with LNR＜30% and≥30% was 3.5% and 9.6% (P=0.018).In age≤35 year eld patients with LNR≥30%,the 5-year RNF and CWR was 40.0% and 20.0%.In T2 patients with LNR≥30%,the 5-year RNF and CWR was 15.8% and 12.2%.Age and LNR were independent prognostic factors for RNF+CWR,LNR was the only independent prognostic factor for CWR by multivariate analysis. Conclusions In T1 or T2 breast cancer patients with 1-3 positive axiliary nodes treated with radical or modified radical mastectomy,a relatively high incidence of chest wall recurrence is observed in the subgroup of patients with lymph nods rate of 30% or greater accompanied by a T2 primary tumor or age≤35 years old. Lymph nodes rate is the only significant prognostic factor of chest wall recurrence.For these patients,post- operative lymphatic drainage area and chest wall irradiation should be considered.

Objective To evaluate the efficacy of port catheter system(PCS)placement in pulmonary artery via percutaneous subclavicle vein treatment for multiple metastatic tumor in the two lungs and discuss the PCS technique.Methods Fifteen multiple metastatic tumor patients(13 hepatocellular carcinomas,one mandible grand adenocarcinoma,one oral bottom squamous carcinoma)were carried out with pulmonary artery PCS placement by way of percutaneous subclavicle vein.FPA/FPM/GP chemotherapy scheme were introduced every 4～6 weeks.Results The success rate of PCS placement technique was 93.3%(14/15).One case failed.Percutaneous subclavicle veins were performed 14 cases in right side and 1 in left one.Following up 2～43 months,2～7 chemotherapy cycles(mean 5 cycles)were accomplished,and the clinical CR and PR were achieved in 1 and 3 cases respectively with clinical efficacy rate 28.6%(4/14).Major side reaction was late wound healing in 1 case.Conclusion PCS placement in pulmonary artery treatment for multiple metastatic tumor in the two lungs is effective,and mastering operation technique is the key for increasing operation suc- cess rate.

OBJECTIVETo evaluate the clinical efficacy and efficacy sustainable time of acupuncture in knee osteoarthritis (KOA).

METHODSThe non-randomized concurrent control trial was adopted. One hundred and ninety-three cases of KOA were divided into an immediate acupuncture group (group A, 97 cases) and a delayed acupunc-weeks at the end of treatment. In group B, the same acupuncture therapy was applied after waiting 4 weeks. The acupoints in the two groups were Liangqiu (ST 34), Dubi (ST 35), Zusanli (ST 36), Yanglingquan (GB 34), Yinlingquan (SP 9), Xuehai (SP 10), Xiyan (EX-LE 4), Xiyangguan (GB 33). WOMAC (Western Ontario and McMasters Universities Osteoarthritis) was used for the assessment of the primary index and VAS (visual analogue scale) was for the secondary index. The evaluation was accomplished by the patients at the beginning of trial, on the 4th and 8th weeks. In each group, 72 patients finished the trial and the data of the lost cases were included in the final data analysis.

RESULTSIn the 4th week of trial, WOMAC score was (25. 8+/-22.0) in group A difference (P<0. 001). VAS scorewas (31. 8+/-24. and was (43.8+/-22.2) in group B, indicating the significant 6) in group A and was (56. 6 +/-25. 8) in group B, indicating very significant difference (P<0. 001). In the 8th week, the efficacy was reduced slightly in the follow-up of group A, but it was improved apparently as compared Acupuncture relieves joint pain and improves joint function obviously.by th patiĩeffr,a Mtaetfti-?an tf ri-with that before treatment.

CONCLUSIONAcupuncture relieves joint pain and improves joint function obviously.The effect of acupuncture is still sustainable in 4 weeks after terminating the treatment.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Arthralgia ; therapy ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; therapy ; Treatment Outcome

OBJECTIVETo investigate the expressions and action mechanisms of nerve growth factor (NGF) receptors TrkA and p75NTR in the oncogenesis and progression of prostate cancer (PCa).

METHODSUsing immunohistochemistry, we detected the expressions of TrkA and p75NTR in 62 PCa and 35 benign prostatic hyperplasia (BPH) samples, and conducted statistical analysis on the basis of clinical data.

RESULTSIndependent-samples t-test showed that, along with poorer tissue differentiation or higher clinical stage of PCa, the expression of TrkA was significantly up-regulated, that of p75NTR remarkably down-regulated, and the expression ratio of TrkA to p75NTR markedly increased. The TrkA/p75NTR ratio was 0.32 in the BPH, 0.52 in the PCa tissue with Gleason score of 6, 1.65 in the PCa tissue with Gleason score of 7, 5.75 in the PCa tissue with Gleason score ≥ 8, 0.89 in the clinical stage of pT2, 1.5 in pT3 a, 3.75 in pT3b, and 7.00 in pTxN1.

CONCLUSIONThe abnormally increased expression ratio of TrkA to p75NTR might be one of the essential features of malignant transformation of prostate cells. A higher TrkA/p75NTR expression ratio may be associated with a lower tissue differentiation, a higher clinical stage or Gleason score, and therefore a poorer prognosis.

Humans ; Immunohistochemistry ; Male ; Neoplasm Grading ; Neoplasm Staging ; Nerve Tissue Proteins ; metabolism ; Prognosis ; Prostatic Hyperplasia ; pathology ; Prostatic Neoplasms ; pathology ; Receptor, trkA ; metabolism ; Receptors, Nerve Growth Factor ; metabolism ; Up-Regulation

To investigate the effect of Tripterygium wilfordii polycoride (TWP) on LPS-induced macrophage inflammatory response, particularly the inhibitory effect on inflammatory factors TNF-α and IL-1β and the regulatory effect on inflammation via TLR4/NF-κB. The MTT method was adopted to test the effects of tested drugs, TWP, dexamethasone (DXM) and azathioprine (AZA) on cell growth to define the appropriate concentration. LPS was used to induce the inflammatory reaction in mouse RAW264. 7 cell lines. The Elisa kit was adopted to test the release level of TNF-α and IL-1β. The Western blotting was applied to test the protein expressions of TNF-α and IL-1β. The RT-PCR was adopted to test the expressions of TLR4 and NF-κB. According to the results, TWP could inhibit the release of macrophage inflammatory factors TNF-α and IL-1β in a dose dependent manner. All of TWP groups showed a weaker efficacy than that of the DXM group. But the TWP high dose group revealed a better effect on TNF-α and equal effect on IL-1β compared with the AZA group. TWP show an equal or better effect in down-regulating TLR4 and NF-κB p65 expressions in a dose dependent manner than DXM and AZA. In conclusion, TWP could inhibit TLR4 and NF-κB p65, which may be related to the down-regulation of TLR4 and NF-κB p65 receptor expressions.
Animals ; Anti-Inflammatory Agents ; pharmacology ; Cell Proliferation ; drug effects ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Inflammation ; drug therapy ; genetics ; immunology ; physiopathology ; Interleukin-1beta ; genetics ; immunology ; Macrophages ; drug effects ; immunology ; Mice ; NF-kappa B ; genetics ; immunology ; RAW 264.7 Cells ; Toll-Like Receptor 4 ; genetics ; immunology ; Transcription Factor RelA ; genetics ; immunology ; Tripterygium ; chemistry

Objective To investigate the in vitro anti-proliferation effect of a histone deacetylase inhibitor,chidamide,on a cutaneous malignant melanoma cell line,A375.Methods Cultured A375 cells were treated with different concentrations of chidamide(5,10,50,100,500 μmol/L)and aichostatin A (TSA)(0.1,0.25,0.5,1.0 μmol/L),respectively,for various durations(24,48,72,96,120 hours).Subsequently,cell proliferation,apoptosis and cell cycle were detected by MTT assay,annexin Vfluorescein isothiocyanate and propidium iodide double staining,and DNA ploid analysis,respectively.Results The proliferation of A375 cells was inhibited in a dose-dependent manner by chidamide of 5-500μmol/L and TSA of 0.1-1 μmol/L,and in a time-dependent manner from 0 to 120 hours after the beginning of trealment with ehidamide of 5-500μmol/L and TSA of 0.25-1μmol/L.The 48-hour 50% growth inhibition concentration(IC50)of ehidamide and TSA on A375 cells was about 250 μmol/L and 0.7μmol/L,respectively.After 48-hour treatment,the apoptosis mte was 80.27%±3.06%,79.53%±5.70%,83.13%±6.90%in A375 cells treated with chidamide of 62.5,125,250 μmol/L,respectively,16.27%±2.46%,28.83%±2.55%,83.40%±8.65%in those treated with TSA of 0.175,0.35,0.7 μmol/L,respectively,10.43%±0.96%in ontreated cells;a statistical increase was noticed in chidamide-treated cells and TSA-treated cells vs.untreated cells(all P＜0.001).A positive correlation was observed between the apoptosis rate and concentrations of TSA(r=0.955,P=0.000).Cell cycle analysis indicated that treatment with chidamide induced cell cycle arrest in G0/G1 phase,with the cell proportion in G0/G1 phase being 76.30%±6.06%,82.79%±0.74%,88.91%±5.29%in A375 cells treated with chidamide of 62.5,125,250μmol/L,respectively,versus 38.73%±3.36%in untreated cells.While after 48-hour treatment with TSA of 0.35 and 0.7 μmol/L,the proportion of cells in G2/M phases was 25.15%±2.71%and 58.71%±3.45%,respectively,compared to 15.73%±0.23%in untreated cells(P＜0.01).Conclusion Chidamide and TSA could induce cell cycle arrest and apoptosis,as well as inhibit the growth of A375 ceils in vitro.