A bacterium that contains a chitin deacetylase has been isolated from ocean soil. It has been identified Baciluus .The best suitable condition of producing this enzyme is that pH is 4.0,35℃,and the incubate time is 80 hours in the present of Ca 2+ as well as having no chitosan. The best temperature of this enzyme when it acts is 40℃～50℃ and the best pH range is 4.5～5.0.

Objective To investigate the effect and mechanisms of recipient-derived immature dendritic cells(imDC) transfected with IKK2dn and loaded with donor antigen on renal allografts survival in the rats.Methods DC were cultured from recipient rats'(Lewis) bone marrow,transfected with IKK2dn and loaded with donor antigen.The expression of CD86 and MHC Ⅱ was detected,and the ability of DC stimulating lymphocyte proliferation in vitro was measured.Male Brown Norwav rats and Lewis rats were used as donors and recipients respectively.Four groups were set up(DC group,empty transfection group,transfection group and control group),receiving 1×10~7 DC,Adv-0-DC,Adv-IKK2dn-DC loaded with BN antigen,and equal volume of normal saline,respectivelv 7 davs before transplantation.In the third party donor-group,Wistar rats as donors were treated the same as DC;group before transplantation.After transplantation,the T lymphocyte proliferation in reciPients was measured and the expression of serum IL-2 and IFN-γ was detected.The survival time of recipients and the acute reiection were observed.Pathological changes were examined tO identify the grade of rejection.Results DC assessment in vivo revealed that the transfected DC could still express CD86 and MHC Ⅱ in a low level as compared with those not transfected with IKK2dn. After DC were loaded with donor's antigen,the expression of CD86 and MHC Ⅱ was up-regulated.After DC were transfected with IKK2dn before loaded with donor's antigen, the expression of CD86 and MHC Ⅱ had no significant change. When DC were loaded with donor's antigen, its allostimulatory activity of T lymphocyte proliferation was enhanced (P<0. 05). When DC were transfected with IKK2dn before loaded with donor's antigen, its allostimulatory activity of T lymphocyte proliferation was not enhanced. Compared with control groups, IKK2dn-transfected DC pulsed with BN splenocyte lysate markedly prolonged the survival of renal allografts (26. 8±1.76d, P<0.01), and elicited markedly lower proliferative responses and reduced IL-2 and IFN-γ production. The pathological grade of rejection was low in the transfection group. Conclusion Recipient-derived imDC transfected with IKK2dn and loaded with donor splenocyte lysate could prolong the renal allograft survival in rats probably by down-regulating the expression of DC costimulatory molecules and inhibiting the T_H 1 cytokine production.

Objective To explore the value of deoxyribonucleic acid(DNA) content on clinical pathological characteristics of nephroblastoma in children.Methods Twenty-four cases of pediatric nephroblastoma from Jan.1993 to Dec.2007 were proved by pathology.The distribution of the 24 cases according to National Wilms Tumor Study group(NWTSG) criteria of USA was stage Ⅰ(12 cases),stage Ⅱ(7 cases),stage Ⅲ(5 cases).The pathology was highly differentiated in 10 cases,medium differentiated in 8 cases and low differentiated in 6 cases according to the level of differentiation.There were 9 cases of lymphatic metastasis and 15 cases of non-lymphatic metastasis in accordance with lymphatic metastasis.The DNA contents of 24 cases were determined by flow cytometry.Chi-square test was used to analyze the relationship among DNA ploidy and clinical stage,level of differentiation,lymphatic metastasis of children with nephroblastoma.Results Aneuploidy mainly occurred in clinical stage Ⅱ or stage Ⅲ(57.14%,100.0%),there was significant difference compared with that of stageⅠ(25.0%)(P

Objective: By observing the differences in Western Ontario and McMaster Universities osteoarthritis index (WOMAC) and visual analog scale (VAS) scores in moxibustion treatment for moderate-to-severe primary knee osteoarthritis (KOA) with moxa of different storage years (3-year moxa and 1-year moxa from Qichun, Huanggang City, Hubei Province, China) through a randomized clinical trial, to objectively evaluate the differences in therapeutic efficacy of moxibustion with moxa of different storage years. Methods: A total of 63 patients with moderate-to-severe KOA who met the inclusion criteria were randomized into moxibustion group 1 and moxibustion group 2 by central randomization method, with 32 cases in moxibustion group 1 and 31 cases in moxibustion group 2. Moxibustion group 1 was treated with moxa stored for 3 years, and moxibustion group 2 was treated with moxa stored for 1 year. Dubi (ST 35), Neixiyan (EX-LE 4) and Heding (EX-LE 2) were selected in both groups, and the treatment lasted 20 min per time, 3 times a week. The immediate efficacy was compared after 6 times of treatment, and long-term efficacy was compared at follow-up 4 weeks after the end of treatment. Results: During the treatment, there were 2 dropouts in moxibustion group 1, and 1 dropout in moxibustion group 2. The total effective rate in the two groups was 83.3％ and 60.0％, respectively. Followed up at 4 weeks after the end of treatment, the total effective rate in the two groups was 80.0％ and 66.7％, respectively. There were no statistical differences between the two groups (both P>0.05). After treatment and 4 weeks after the end of treatment, the WOMAC and VAS scores in both groups decreased significantly compared with those before treatment (all P<0.01); the scores of stiffness item of WOMAC in moxibustion group 1 were lower than those in moxibustion group 2 (both P<0.05); there were no statistical differences in the scores of pain item and dysfunction item of WOMAC, and VAS scores between the two groups (all P>0.05). Conclusion: Moxibustion with moxa of different storage years (stored for 3 years and 1 year) both can improve the pain, stiffness and motor function in patients with moderate-to-severe KOA. While moxa stored for 3 years has a better therapeutic efficacy in improving stiffness of the knee joint than that stored for 1 year.

OBJECTIVETo investigate the relationship between p53 gene codon 72 polymorphism and genetic predisposition to keloid in Chinese population.

METHODSPCR-based restriction fragment length polymorphism (PCR-RFLP) analysis was used to detect p53 gene codon 72 genotypes of 60 keloid samples and 102 whole blood samples from healthy controls in China.

RESULTSThere was no significant difference in the distribution of p53 gene codon 72 polymorphism between the keloid patients and the healthy controls (X2 = 2.910, P = 0.233), nor did the frequencies for Pro and Arg alleles (X2 = 0.882, P = 0.348), and there was no significant difference in the distribution of p53 gene codon 72 polymorphism in keloid patients and normal controls from China and Japan respectively (X2 = 3.942, P = 0.139; X2 = 3.260, P = 0.196). But the Arg/Arg genotype was significantly higher than the Pro/Pro genotype among the patients with keloid in shoulder and back (P < 0.01).

CONCLUSIONSThere was no significant association between the distribution of p53 gene codon 72 polymorphism and keloid in Chinese population, but Arg/Arg genotype may affect the formation of keloids in shoulder and back compared to others. Further research should be done to investigate the relationship between p53 gene codon 72 polymorphism and keloids in different sites.

Asian Continental Ancestry Group ; genetics ; Codon ; genetics ; Female ; Humans ; Keloid ; genetics ; Male ; Polymorphism, Restriction Fragment Length ; Tumor Suppressor Protein p53 ; genetics

AIM:To explore the application of 10g/L cyclopentolate chloride eye drops in children, and to compare the different effectiveness of cycloplegia between 10g/L cyclopentolate chloride and atropine in Chinese children.METHODS:A total of 236 eyes of 118 children aged 3~12 years old were enrolled in this study including 80 eyes of 40 children with myopia, 156 eyes of 78 children with hyperopia and 146 eyes of 73 children combined with astigmatism. 10g/L cyclopentolate chloride eye drops were used once per 5min for 3 times and refractive diopter was obtained 1h after the last drop of cyclopentolate. Three days after that, 10g/L atropine was then used 1 time per night for 1wk and optometry was performed again. The children were divided into 3 groups ( myopia, hyperopia and astigmatism group ) according to the refractive status, in which astigmatism was independent of the degree of separation of cylinder statistics. The results of retinoscope refraction were then compared between 10g/L cyclopentolate and 10g/L atropine.
RESULTS:The refractive diopter was -2. 25±1. 31D after 10g/L cyclopentolate eye drops and -2. 23±1. 32D after 10g/L atropine in myopic group. The refractive diopter was 1. 35±1. 19D and 1. 38±2. 00D in astigmastic group. No significant difference was found in myopic group and astigmastic group (P= 0. 109, P= 0. 374). While in the hyperopic group, the refractive diopter was 3. 76±2. 4D after 10g/L cyclopentolate eye drops, which was lower than that after 10g/L atropine 4. 39±2. 6D (P=0. 000).
CONCLUSION: The results of this study suggest that 10g/L cyclopentolate chloride eye drops can be used in myopia and astigmatism children, and 10g/L atropine should be used in hyperopia children.

Objective To explore the relationship of proteinuria,immunopathogenesis and humoral immunity in children with Henoch-schonlein purpura nephritis(HSPN).Methods Serum IgG,IgA,IgM,C3 and C4 levels were detected with complete-automatic biochemistry analyzer in 47 children with HSPN and 18 heathy children,and the differences in the findings were compared among groups.The levels of IgG,IgA,IgM,C3 and C4 were compared among groups according to the count of proteinuria.The renal tissues were adopteded with immunity in the children with HSPN were taken for biospy examination,and the differences in immunopathalogy were compared among groups with immunof-luorescence assay.Results The IgG,IgA levels in children with HSPN were higher than those of heathy children,on the contrary,the IgM,C3 and C4 levels were lower than those of heathy children(Pa0.05),but the IgM level of the nephritic syndrome group were higher than that of other groups(Pa0.05).Conclusions There is severe disturbance of immunologic function in children with HSPN.There is no connections among serum IgG,IgA,IgM,C3 and C4 levels,proteinuria and renal immunopathlogy.

OBJECTIVETo study the effect of ursolic acid (UA), apentacyclic triterpene acid, on MCF-7 cell apoptosis, and probable mechanism involved by detecting the expressions of caspase-3 and poly ADP-ribose polymerase(PARP) at protein level.

METHODMCF-7 cells were cultured with different concentrations of UA. Growth inhibition of UA on MCF-7 cells was evaluated by MTT assay. Cell cycle and sub-G1 peak were performed by FCM. Morphologic changes of UA-treated cells were observed by light microscope. Apoptotic cells with condensed or fragmented nuclei were visualized by Ho 33258 staining by a fluorescence microscope (EX: U. V.). The protein expression of caspase-3 and PARP was analyzed by immunofluorescence cell staining (SABC-Cy3).

RESULT24 hours after UA treatment, inhibition of MCF-7 cell growth was concentration-dependent. The IC50 value for UA was (22.6 +/- 3.0) micromo x L(-1). Cell cycle anaysis by FCM showed that 50 micromol x L(-1) of UA arrested MCF-7 cell cycle at G0 - G1 phase. Morphological changes of MCF-7 Cells exhibited many of the hallmark features of apoptosis, including chromatin clumps and aggregation and DNA fragmentation. UA increased caspase-3 protein expression.

CONCLUSIONThe results suggest that UA evokes MCF-7 cell apoptosis is correlation with the up-regulation of caspase-3. Our study indicated that UA might be a potential Chinese medical component for breast neoplasm.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Breast Neoplasms ; enzymology ; pathology ; Caspase 3 ; Caspases ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Female ; Humans ; Poly(ADP-ribose) Polymerases ; metabolism ; Triterpenes ; pharmacology

Choosing the right solvent and timely use is the basis of rational drug use and the most direct and efficient way to improve the safety of traditional Chinese medicine injections. This study aimed to evaluate the influence of solvent and drug preparation time on Shuanghuanglian injection inducing pseudo-allergic reactions with mouse mode. The two tests were carried out: (1) Comparative experiment between different solvent: Shuanghuanglian injection preparation to the appropriate concentration with 0.9% sodium chloride injection and 5% dextrose injection, mixed with Evans blue, at one time intravenous injected into mice, 30 minutes later, the mouse ears vascular permeability were observed and compared. (2) Comparative experiment among different preparation time: placed 10 min, 2.5 h, 6 h and 24 h after Shuanghuanglian injection were prepared and then to detect the pseudo-allergic reactions in mice using the same methods as in (1). The results showed that there was no significant difference in the pseudo-allergic reactions in mice which induced by the same dose of Shuanghuanglian injection, respectively with 0.9% sodium chloride injection and 5% dextrose injection preparation, and with the extension of preparation time, the degree of pseudo-allergic reactions of Shuanghuanglian injection was gradually severe.
Animals ; Drug Compounding ; Drug Hypersensitivity ; etiology ; Drugs, Chinese Herbal ; adverse effects ; Injections ; Male ; Mice ; Mice, Inbred ICR ; Solvents ; Time Factors

To evaluated the pseudo-allergic reactions of cordate houttuynia, pulse-activating and Qingkailing injection in mice, the ICR mouse were divided into different test groups, then were intravenously injected with three traditional Chinese medicine injections, positive control compound 48/80 and physiological saline as normal control. All test substances were mixed with 0.4% Evans blue. The reaction and vascular permeability of the ears were observed and measured 30 min after injection. At the same time, the mechanisms of the traditional Chinese injections' pseudo-allergic reactions was studyed. ICR mice were injected with the test substances as above without EB, blood in a part of mice were collected after 5 min, and the level of histamine in the plasma were measured. Blood in the other part of mice were collected after 30 min, and the level of VEGF, TNF-α and IL-10 in the serum were measured. The reasults showed that except the cordate houttuynia injection, pulse-activating injection in 1. 5 times clinical concentration and Qingkailing injection in 3.3 times clinical concentration caused mild pseudo-allergic reactions mainly for vascular permeability, no pseudo-allergic reactions occurred when the concentration of the two injections was below the concentration metioned above; 5 minutes after intravenous injection of the three TCM injections into ICR mice with the highest dose, the levels of histamine in plasma of pulse-activating injection and Qingkailing injection groups were increased significantly, 30 minutes later, the levels of VEGF, TNF-α and IL-10 in the serum of all groups were no significant change compared to normal group. The mouse of pulse-activating and Qingkailing injection groups showed inflammatory changes in ear and lung tissues. It can be conluded that when the dose or concentration increased to a certain extent, pulse-activating and Qingkailing injection could induce pseudo-allergic reactions on ICR mice, the increased histamine realease maybe the main reason for pseudo-allergic reactions of the two traditional Chinese medicine injections. In addition the author preliminary thought that inflammatory mechanisms leading to hyperpermeabilities were the main reason of the traditional Chinese medicine injection's pseudo-allergic reaction.
Animals ; Drug Hypersensitivity ; etiology ; Humans ; Injections ; Interleukin-10 ; blood ; Male ; Medicine, Chinese Traditional ; adverse effects ; Mice ; Mice, Inbred ICR