Objective To establish the murine animal model for acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation (HSCT). Methods T cell-depleted (TCD) BM cells from allogeneic C57BL/6 mice were harvested and prepared from the marrow of the femurs and tibias. Age of 4 to 6 weeks’ BALB/c mice received 900 cGy total body irradiation ( TBI) from a 137Cs source. Two to four hours later,the mice were injected intravenously ( i. v. ) 5 × 106 TCD-BM cells with 1. 25 × 106 or 2. 5 × 106 or 5 × 106 splenic cells from allogeneic C57BL/6 mice,and respectively divided them into experimental group 1,experimental group 2 and experimental group 3. TCD-BM only was used as the negative control group of GVHD. Ten mice were used for each group. The establishment of aGVHD model was evaluated with a clinical GVHD scoring system,survival rate and target-organ damage. Results On 7 to 14 days after transplanta-tion,the typical clinical GVHD manifestation of severe diarrhea,hogback,activity reduced and ruffling were observed in experimental group 2. Furthermore,the aGVHD target organs of colon,lung and liver were harvested and made histological paraffin sections,then the obviously path-ological tissue damages of GVHD were detected under microscope. And the survival rate was lowed down to 0 on 45 days after transplantation in experimental group 2. On the contrary,no obviously clinical manifestation of aGVHD were observed in the control group,experimental group 1 and group 3. On 60 days later, the survival rate was 80% in experimental group 1 and 100% in control group. However,no mice was sur-vived on 10 days later in group 3. Conclusion BALB/c mice aGVHD model after allogeneic HSCT is successfully established by injecting i. v. 5 × 106 TCD-BM cells with 2. 5 × 106 splenic cells from allogeneic C57BL/6 mice.

OBJECTIVETo observe the effect of Polygonatum sibiricum on Yin deficiency model rats induced by long-term overload swimming.

METHODExcept for the normal group, all of the remaining rats performed the long-term overload swimming for eight weeks, with five days every week and once every day, to establish the Yin deficiency model. The daily swimming time increased from 10 min to 180 min at the end of the 7th week, with the water depth of 60 cm and the water temperature at 30 degrees C. After the success of the modeling, the rats were orally administered with different doses of aqueous extracts from P. sibiricum (2.5, 10 g x kg(-1)) for eight weeks. After the final administration, their blood were collected from orbits to measure immunoglobulin A, G and M (IgA, IgG, IgM), interleukin 2 and 6 (IL-2, IL-6) and cAMP, cGMP contents in plasma General behavioral indicators (weight, facial temperature, pain threshold and holding power) of rats were observed during the drug administration.

RESULTCompared with the model control group, aqueous extracts from P. sibiricum was given for eight weeks to significantly increase the rat weight and holding power of Yin deficiency model rats, decrease the facial temperature and the sensitivity of pain threshold, and increase IgA, IgG, IgM and IL-6 content and IgG content in serum, but without statistical difference. Aqueous extracts from P. sibiricum (10 g x kg(-1)) could also increase IL-2 content in serum, and decrease cAMP content and cAMP/cGMP ratio.

CONCLUSIONP. sibiricum could improve the general behavioral indicators (weight, holding power, pain threshold and facial temperature), immunologic functions (IgA, IgG, IgM) and cyclic nucleotide (cAMP, cAMP/cGMP), so as to ameliorate such Yin deficiency symptoms as dysphoria in chestpalms-soles, weight loss, soreness and weakness of waist and knees, immunologic dysfunction and cyclic nucleotide system disorders.

Animals ; Body Weight ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Polygonatum ; chemistry ; Rats ; Rats, Sprague-Dawley ; Swimming ; Yin Deficiency ; drug therapy ; physiopathology

Objective To establish RT-PCR-RFLP method for studying the genotype of wild mea-sles virus strains isolated from Tianjin area from 2002 to 2008. Methods Isolations of measles virus were carried out by tissue culture method from urine and throat swab specimens collected from suspected cases. RNA were extracted from the virus specimens. The 594 bp fragment of C terminal of the N (nucleoprotein) gene was amplified by one-step RT-PCR, then the PCR products were digested with Bcn I , separated on agarose gel electrophoresis and then analyzed by the method of RFLP (restriction fragment length polymor-phism). In addition, above results were compared with DNA sequencing. Phylogenetic tree was plotted based on the results for the genetic relationship and distance analysis. Results Sixty-nine measles virus strains were isolated from 189 specimens from 2002 to 2008, of which the C terminals of N gene were all de-tected positive. Among the 69 strains of measles virus isolates, 98.55% (68/69) belonged to Hla sub-geno-type which was the predominant sub-genotype, and only one strain (1.45%) belonged to H1b sub-genotype by RFLP analysis which was in accordance with the results by DNA sequencing method. Phylogenetic tree analysis indicated the H1a sub-genotype measles virus strains should be further divided into 2 clades, and the variation fluctuated between 0.2% and 3.8%. There were transmission chains caused by different virus strains co-cireulation. Conclusion A genotype, H1a and H1b sub-genotype can be identified by RT-PCR-RFLP assay specically based on the restriction enzyme Bcn I .The RT-PCR-RFLP assay can be a rapid, simple, accurate and efficient method for large-scale surveillance of measles virus strains in China.

Objective To study whether the organs besides digestive system of musca domestica Ⅲ instar larvae have the capability of produceing musca β-glucosidase.Methods Tissues of malpighian tubules,trachea,epiploon and body wall of musca domestica Ⅲ instar lar-vae were dissected under anatomic microscope,and the expression of β-glucosidase gene in these dissected tissues were detected by reverse transcription PCR.And the tissue localization of β-glucosidase mRNA was further identified by in situ hybridization.Moreover,anti-cellulase was used to determinate the tissue distribution with immunohistochemical staining.The relative mRNA expression levels of musca domesticaβ-glucosidase gene in these organs were tested by real-time quantitative PCR.Results The reverse transcription PCR showed that the ampli-fication products of β-glucosidase gene were observed in tissues of malpighian tubules,trachea and body wall.β-glucosidase mRNA was shown in the epithelium cells of malpighian tubules,trachea and body wall by in situ hybridization,and it was almost the same in the results of im-munohistochemical staining.The real-time quantitative PCR showed that the relative expression quantity of β-glucosidase gene in malpighian tubules and body wall were higher than that in foregut,while it was lower in itrachea than that in foregut.And it was of statistical difference in gene expression level of β-glucosidase among these organs (P <0.05).Conclusion Malpighian tubules,trachea and body wall of musca domestica Ⅲ instar larvae have the function of secreting β-glucosidase.Combining with the characteristics of secreting β-glucosidase in most organs of digestive system,it may provide a new biological method for the prevention and treatment of human diseases transmitted by musca domestica with the use of taget gene β-glucosidase.

Objective To study the diagnostic value of pencilliosis marneffei (PM)in a non-HIV-infected child with the com-bined detection of aspergillosis galactomannan,fungus Glucan(1-3)-β-D and boold culuture.Methods The venous blood specimen from the child was collected for the quantified detection of aspergillosis galactomannan,fungus Glucan(1-3)-β-D. The growth and colonial morphology of fungus was inspected with the positive blood culture and the characteristics of fun-gus smear were observed under microscope.Results The result of aspergillosis galactomannan was 14.45 μg/L and fungus Glucan (1-3)-β-D 77.14 pg/ml.Penicillium marnrffei was identified using blood culture.It was mycelia form under 25℃ and the salouraud medium produced water soluble claret-red pigment produced.It was mycelia form under 35℃ and the colony was gyri creases,the characteristic broom-like hypha and separation hypha could be found under microscope.Conclusion It is effective for the early diagnosis and therapy of PM with the combination detection of aspergillosis galactomannan,fungus Glucan (1-3)-β-D and boold culuture and have better clinical diagnosis value.

Objective To investgate the effect of nervous growth factor(NGF) on the proliferation of the limbal stem cells(LSCs) in vitro,and the relationship bewteen expression of its receptors and cell proliferation.Methods After primary cultured,LSCs were divided into the control group and the NGF group.Selected cells cultured of 1 d,3 d and 5 d in the two groups and examined the expression of p63,TrkA,p75 with immunohistochemistry.Results The average gray scale values of expression of p63,TrkA and p75 at 1 d,3 d and 5 d in NGF group were significant decreased compared with the corresponding data in the control group(P<0.05).Pearson's correlations analysis showed that the average gray scale values of expression of TrkA and p63 were of statistically significant differences(P<0.05).Conclusion These results highlight that NGF could maintain the stem cell properties of LSCs.LSCs could exepress the NGF receptors of TrkA and p75,and the expression of TrkA showed a correlation with LSCs proliferation.

Objective To determine the expression of HPV16/18,31/33 DNA and p53,p21~(WAF1) and MDM2 proteins in invasive squamous cell carcinoma of cervix (ISCC)and to indicate the significance of them in the occurrence and development of ISCC.Methods Using tissue microarray,in situ hybridization (ISH) and immunohistochemical method,we detected the expression of HPV16/18,31/33 and investigate the expres- sion of p53,p21~(WAF1),MDM2 and proteins in ISCC,CIN and NCE.Results was analysed by SPSS vision 12.5. Results The positive expression rate of HPV16/18,p53,p21~(WAF1),MDM2 in ISCC was markedly higher than in CIN and NCE.We found the difference between HPV31/33 and lymph node transfer.Significant relation- ship was observed between p53 protein expression and histological grade and lymph node metastasis of the cancer.There was positive correlation between the expression of p21~(WAF1) protein and the depth of invasion(P

Objective To examine how stroke affects muscle coordination and whether muscle function will be improved after rehabilitation.Methods Ten stroke patients with mild hemiparesis and six age-and sex-matched controls were investigated at baseline.Velocity-encoded phase-contrast magnetic resonance imaging (VE-PC MRI) and surface eleetromyography (sEMG) were performed to evaluate muscle coordination of thigh muscles during knee extension and flexion and the effect of rehabilitation. Results Using VE-PC MRI,we found that the peak velocity of rectus femoris was lower in the affected limb (P

Objective To find the effective therapeutic arrangement through investigating the clinical characteristics of chronic cardiac insufficiency with anaemia. Methods A total of 46 cases of anemia from 315patients who had been admitted to department of cardiology, the First Affiliated Hospital, Harbin Medical University for chronic cardiac insufficiency with anaemia were selected. They were divided into two groups. There were 22 patients in the first group who only accepted treatment to improve cardiac function (normal cardiac, diuretic,vasodilator therapy, etc.), and 24 patients in the second group who accepted treatment to improve cardiac function while receiving anti-anemia therapy treatment, oral ferrous sulfate tablets(0.3 g/tablet), 1 tablet each time, 3 times a day and(or) 2 times per week subcutaneous erythropoietin 3000 U. The hemoglobin(Hb), red blood cell(RBC) ,hematocrit (HCT), left ventricular ejection fraction (LVEF), fractional shortening (FS), stroke volume (SV) , cardiac output(CO) and E peak and A peak ratio(E/A) were observed before and after treatment. By logistic regression, grade grade Ⅱ , Ⅲ , Ⅳ, the incidence of anaemia were 7.9% (10/126), 19.2% (23/120) and 24.6% (17/69),respectively. Grade Ⅱ compared with grades Ⅲ, Ⅳ, the difference was statistically significant (x2 = 4.08, 3.12, all (3.49 ± 0.17) × 1012/L, (0.36 ± 0.08)%, (48.9 ± 3.11)%, (15.6 ± 1.8)%, (38.9 ± 3.7)%, (4.4 ± 1.6)% and (130.7 ±5.75)g/L, (4.12 ± 0.25) × 1012/L, (0.43 ± 0.02)%, (58.5 ± 2.65)%, (18.0 ± 2.5)%, (49.1 ± 7.7)%, (5.1 ± 1.2)%in the first and second groups, respectively. The difference between the two groups was statistically significant(t =value of Hb, RBC, HCT, LVEF, FS,SV, CO were (102.7 ± 6.93)g/L, (3.41 ± 0.12) × 1012/L, (0.35 ± 0.07)%,(47.5 ± 2.86)%, (16.0 ± 2.4)%, (38.2 ± 7.9)%, (3.7 ± 1.4)%, respectively. Compared with those after treatment,the difference was statistically significant (t = 15.632, 13.325, 5.569, 17.182, 3.186, 2.999, 3.074, all P ＜ 0.05);Ⅳ-level relative risk were 1.62, 3.14(P ＜ 0.05 or ＜ 0.01) . Conclusions Based on the standard treatment with treatment of anemia, cardiac contractile function can be improved.

ObjectiveTo study tumor necrosis factor-related apoptosis-inducing ligand(TRAIL) 1525 locus polymorphism in patients with nodular thyroid disease and investigate the relation between individual gene polymorphism and thyroid diseases.Methods A total of 125 patients were diagnosed with nodular thyroid disease at the Department of Endocrinology,the First Affiliated Hospital of Inner Mongolia Medical College.Among these patients,67 cases were nodular thyroid goiter and 58 cases were nodular thyroid adenoma,54 males,71 females,and average age was 41.05 ± 14.42. Patients with nodular thyroid goiter were grouped into toxic and non-toxic and thyroid adenoma were grouped into high-functioning or non-high-functioning.A total of 100 healthy subjects.47 males,53 females,average age 42.35 ± 16.52 were as control group.According to the principle of informed consent,venous blood was collected,polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for TRAIL gene 1525 locus polymorphism determination was performed to calculate genotype of the TRAIL gene (homozygous GG,heterozygous GA,mutationhomozygous AA) and the gene frequency (G,A),and relative degree of risk(odd ratio,OR) was compared.ResultsNodular goiter group TRAIL gene 1525 locus genotype frequencies(GG:40.3％,AG:44.8％,AA:14.9％),allele frequencies(G:62.7％,A:37.3％) were compared with that of the control group(GG:17.0％,AG:65.0％,AA:18.0％; G:49.5％,A:50.5％),the differences were statistically significant(x2 =11.376,5.633,P ＜ 0.01 or ＜ 0.05 ).Adenomas group 1525 locus genotype frequencies of the TRAIL gene(GG:44.8％,AG:38.0％,AA:17.2％),allele frequencies(G:63.8％,A:36.2％) were compared with that of the control group,the differences were statistically significant(x2 =15.342,6.054,P ＜ 0.01 or ＜ 0.05).Allele frequencies of thyroid goiter group and denoma group were compared with that of the control group,OR values were 1.714 and 1.797(all P ＜ 0.05) and 95％ confidence intervals were 1.097 - 2.679 and 1.124 - 2.874.The difference of 1525 locus genotype or allele frequency distribution in toxic and non-toxic nodular group,high functioning and non-high-functioning adenomas group were not statistically significant (x2 =3.714,2.792; 1.103,2.020; all P ＞ 0.05).ConclusionTRAIL gene 1525 locus polymorphism is significantly associated with nodular thyroid disease.