We have found the parallel relationship between the live cell number of several tumor cell lines and the formation of MTT formazan with MTT colorimetric assay. The MTT colorimetric assay was compared with ~3H-Tdr incorporation assay for the proliferation of mouse spleen cells induced by mitogens or the activity of rat/mouse IL2,the results suggested that the ~3H-Tdr incorporation assay can be replaced by MTT colorimetric assay. MTT colorimetric assay have the advantages of simplicity, rapidity,save any radioisotope and no specific equivepment, etc, so under some conditions it would be a useful method for measuring cell proliferation or cytotoxicity in a laboratory.

OBJECTIVETo study the effects of Shibao Powder on the expression of b-FGF and TGF-beta 1 in granulation tissues of rabbit models with soft tissue injuries, so as to explore mechanism of external Chinese drugs for repairing of soft tissues in molecular levels.

METHODSThe rabbit models were established by classical method of full-thickness skin wounds. After 8% sodium sulfide was used and routine disinfection completed, intra-peritoneal anaesthesia was adopted. The skin was cut to expose gastrocnemius muscle, and imcomplete sharp dissection was made near the tendon insertion. The length of the incision was 0.8 cm and the width was 0.4 cm. The rabbits in the experimental group were treated with spread of Shibao Powder at the wound; the rabbits in clean group were treated with routine dressing changes and disinfection; and the rabbits in the control group were treated with dressing changes only. The granulation tissues in different stages were collected and observed with high power microscope. The expression of Transforming Growth Factor-beta 1 (TGF-beta 1) protein and b-FGF protein in wound tissues were detected using StreptA-ridin-Biotin-Complex (SABC) method.

RESULTSThe results showed that the expression of b-FGF protein had significant differences among 3 groups at the 6th day after trauma (P<0.05). The TGF-beta 1 protein expression also had significant differences among 3 groups at the 10th and 14th days after trauma (P<0.05). At the same time, the growth states of granulation tissues had difference among 3 groups at the same trauma stage, and within the same group among different trauma stages.

CONCLUSIONThis experimental study shows that Shibao Powder is effective to promote the repair of soft tissues after trauma by stimulating production of endogenous growth factor from cells in wound.

Animals ; Disease Models, Animal ; Fibroblast Growth Factors ; metabolism ; Gene Expression Regulation ; drug effects ; Granulation Tissue ; drug effects ; metabolism ; pathology ; Male ; Medicine, Chinese Traditional ; Powders ; Rabbits ; Soft Tissue Injuries ; drug therapy ; metabolism ; pathology ; Transforming Growth Factor beta1 ; metabolism ; Wound Healing ; drug effects

This study was aimed to investigate the effect of rhIL-6 and rhEPO on hepcidin mRNA expression in HepG2 cells and human primary hepatocytes, and mechanism of rhEPO in treatment of anemia of chronic disease (ACD). The HepG2 cells and human primary hepatocytes were cultured with medium containing different concentrations of rhIL-6 and rhEPO for a certain time, then mRNA was isolated and its RT-PCR was performed, the bands were photographed and analyzed by UVI band, the hepcidin and G3PDH mRNA ratio were semi-quantitatively analyzed. The expression levels of hepcidin in GepG2 cells and human primary hepatocytes at different conditions were compared. The results showed that the hepcidin mRNA expression in HepG2 cells and human primary hepatocytes could be enhanced by rhIL-6, the rhEPO could inhibit rhIL6-induced hepcidin mRAN expression. The rhEPO alone basically did not influence hepcidin mRNA expression in HepG2 cells. It is concluded that Hepcidin mRNA expression in HepG2 cells and human primary hepatocytes can be elevated by rhIL-6 with concentration- and time-dependent manner in certain range. rhEPO can inhibit this effect of rhIL-6.
Antimicrobial Cationic Peptides ; genetics ; metabolism ; Erythropoietin ; pharmacology ; Hep G2 Cells ; Hepatocytes ; drug effects ; metabolism ; Hepcidins ; Humans ; Interleukin-6 ; pharmacology ; RNA, Messenger ; genetics ; Recombinant Proteins ; pharmacology

Aged ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; immunology ; Prognosis ; T-Lymphocyte Subsets ; immunology

OBJECTIVETo evaluate the clinical value of the comprehensive therapy of acupuncture, Chinese herbal medicine and rehabilitation in the treatment of post-stroke flaccid limb dysfunction.

METHODSThe four-center, single-blind, randomized and controlled research method was adopted, 240 qualified subjects were randomized into a comprehensive therapy group, an acupuncture group, a rehabilitation group and a Chinese herbal medicine group, 60 cases in each one, at the ratio of 1 1. In the comprehensive therapy group, the comprehensive therapy of acupuncture, Chinese herbal medicine and rehabilitation was applied. The acupuncture therapy included the scale acupuncture at middle line of vertex, lateral line 1 of vertex, lateral line 2 of vertex, etc. with the single reinforcing and reducing technique by the speed of needle insertion and withdrawal, and the body acupuncture therapy at the acupoints on the antagonistic muscles with the reinforcing and reducing technique by the needle rotation. The Chinese herbal medicine therapy included No. 1 stroke formula for the cases of liver and kidney yin deficiency and the upward disturbance of wind yang, No. 2 stroke formula for qi deficiency and blood stagnation, and the stagnation in meridians and No. 3 stroke formula for the interaction of phlegm and stasis and blockage of meridians according to the pattern/syndrome differentiation. The rehabilitation therapy focused on the promotion technique by putting the healthy limb. The simple acupuncture, rehabilitation and Chinese herbal medicine therapies as the comprehensive therapy group were applied in the acupuncture group, rehabilitation group and Chinese herbal medicine group separately. The Chinese medicine symptom, the limb motor function, the daily life activity, fainting needle reaction, allergic reaction and the others were taken as indices to evaluate the efficacy and safety of the treatment.

RESULTS(1) The results of the four indices named the Chinese medicine symptom, the limb motor function, the limb balance function, the daily life activity were all improved significantly after treatment as compared with those before treatment in four groups (all P < 0.01). (2) Concerning to the improvement degrees, the improvements of the above four indices in the comprehensive therapy group were more significant than those in the other three groups (P < 0.01, P < 0.05). The improvement in Chinese medicine symptom in the acupuncture group and the Chinese herbal medicine group were more significant than that in the rehabilitation group (both P < 0.05). The improvement of the upper limb motor function in the acupuncture group was more significant than that in the rehabilitation group and the Chinese herbal medicine group separately (both P < 0.05).

CONCLUSIONThe comprehensive therapeutic program of acupuncture, Chinese herbal medicine and rehabilitation is safe and effective in the treatment of post-stroke flaccid limb dysfunction. It is more advantageous in efficacy as compared with any simple therapy.

Activities of Daily Living ; Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Muscle Hypotonia ; drug therapy ; etiology ; rehabilitation ; therapy ; Stroke ; complications

To investigate the effect of EPO on hepcidin mRNA expression in normal mice, acute inflammatory mice and ACD mice, the acute phase model and ACD model of mouse was produced by subcutaneous injection of turpentine oil. Hepatic hepcidin mRNA expression levels of normal mice, acute inflammatory mice and ACD mice were detected by semi-quantitative retro-transcriptase polymerase chain reaction (RT-PCR) after intra-peritoneal injection of EPO. The results showed that prolonged chronic inflammation did not significantly increase mouse hepatic hepcidin mRNA expression, but a single injection of turpentine oil increased mouse hepatic hepcidin mRNA expression transiently. Intra-peritoneal injection of EPO down-regulated hepatic hepcidin mRNA expression in normal mice, ACD mice and acute inflammatory mice. Hb levels had a negative correlation with hepatic hepcidin mRNA expression levels in ACD mice treated with EPO. It is concluded that the ACD model can be produced by repeated subcutaneous injection of turpentine oil. Hepatic hepcidin mRNA expression increased during the early period of ACD process and then fall to normal level. EPO down-regulate hepcidin mRNA expression under the normal, acute inflammation and chronic inflammation conditions.
Anemia ; chemically induced ; metabolism ; Animals ; Antimicrobial Cationic Peptides ; biosynthesis ; genetics ; Erythropoietin ; pharmacology ; Hepcidins ; Inflammation ; chemically induced ; metabolism ; Iron ; metabolism ; Liver ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation

Objective To explore the distribution of water with high level arsenic and prevalence of arsenism along Huai'he River and the surrounding area of Hong'ze lake in Huai'an of Jiangsu. Methods Wate rsamples were collected and tested in 2008 from 18 villages of 6 towns according to history data in 3 counties like Xuyi,Jinhu and Hongze. Samples having arsenic level higher than 0.05 mg/L were investigated by epidemiological method and the patients were diagnosed by Standard of Diagnosis for Endemic Arsenism. Results All 5199 water samples were determined,and 260 water samples were exceeding the national drinking water quality level (0.05 mg/L) in 3 counties,the rates of exceeding diagnosis were 5.6%(247/4454),0.7%(4/597),6.0%(9/148) respectively. Total detected rate of endemic arsenic disease was 5.94%(128/2155). The detected rates of age group of 0 ～ ,20 ～,30 ～ ,40 ～ ,50 ～ ,60 ～ ,70 ～ ,80 ～ were 2.86%(1/35),2.11%(2/95),1.26%(3/239),3.10%(16/516),5.53% (32/579),10.07%(41/407),11.84%(27/228),10.71%(6/56) respectively. The detected rate of male (9.10%,78/857) was higher than that of female(3.85%,50/1298,χ~2 = 25.46,P < 0.01). Conclusions Huai'he River and the surrounding areas of Hong'ze lake like Xuyi,Jinhu and Hongze are identified existing endemic arsenic disease area. The prevention of arsenism should be strengthened in these areas.

OBJECTIVETo evaluate the efficacy and safety of high-dose dexamethasone-based regiments in newly diagnosed multiple myeloma patients with renal impairment.

METHODSThe clinical data of 22 patients with newly diagnosed multiple myeloma patients with renal impairment who received high-dose dexamethasone-based regiments from August 2006 to August 2008 in Peking Union Medical College Hospital were retrospectively reviewed.

RESULTSAfter receiving a median 4 cycles of high-dose dexamethasone-based regiments, renal impairment was reversed in 7 patients (31.8%) with a median time to reversal of 31 days. Sixteen patients (72.7%) achieved overall response, including 7 patients (31.8%) had complete remission / near complete remission. The grade 3 or 4 adverse events included neutropenia (13.6%), infections (22.7%), peripheral neuropathy (9.1%), and ileus (4.5%).

CONCLUSIONThe high-dose dexamethasone-based regiments are safe and effective for newly diagnosed multiple myeloma patients with renal impairment.

Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Dexamethasone ; administration & dosage ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; complications ; drug therapy ; Prognosis ; Renal Insufficiency ; complications ; Retrospective Studies ; Treatment Outcome

Objective To investigate the feasibility,efficacy and safety of high dose chemotherapy (HDC)and autologous peripheral blood stem cell transplantation(PBSCT)with CD34~+ cell selection in patients with severe primary Sjgren's syndrome(pSS).Methods Three patients with persistent and severe pSS de- spite of glucocorticoid and immunosuppressive treatment were enrolled from 1999.All patients underwent high dose chemotherapy and peripheral blood stem cell infusion with CD34~+ cell selection.Autologous hematopoietic stem cells were mobilized with 2～3 g/m~2 CTX(total dosage,infused in two days)and 5?g/kg granulocyte colony stimulating factor(G-CSF),enriched with CD34~+ cell selection by CliniMACS,and reinfused after con- ditioning with 200 mg/kg CTX and 90 mg/kg swine antithymocyte globulin or 200 mg/kg CTX and total body irradiation of 4 Gy.Results One patient got complete remission after 2 times of mobilization,so no condi- tioning and transplantation were given.Other 2 patients completed the mobilization and leukapheresis proce dures successfully,and proceeded to receive conditioning and transplantation.All patients had rapid hematopoi- etic reconstitution.Three patients were followed up for 48 months,60 months and 18 months,respectively.All patients were free from abnormal activity of B lymphocytes.The titer of antibody decreased and anti-SSB anti- bodies of 2 patients turned to negative.Aggregation of focal lymphocytes in labial gland disappeared in 1 pa- tient after PBSCT.Improvement of pulmonary dysfunction and reversibility of interstitial pulmonary fibrosis fol- lowing hematopoietic stem cell transplantation were observed in 2 patients.Conclusion The abnormal activity of B lymphocytes in pSS patients can be controlled successfully with PBSCT.High close chemotherapy followed by peripheral blood stem cell transplantation with CD34~+ cell selection is feasible and safe.

To establish a new high resolution melting analytical method for identification of Lonicera japonica germplasm, the screening of 7 pairs of SSR (simple sequence repeats) primers, determining the suitable diagnostic primers by the differences of peak pattern and Tm was conducted. Then into the DNA template concentration, annealing temperature and the suitable range of cycle number were investigated. Combined with SIMCA-P software for data processing analysis, the results show that three main germplasm honeysuckle could be divided by four sets of primers. It provides methodology for improving L. japonica germplasm identification.