Objective To provide clinical evidence for the relationship between epithelial-mesenchymal transition (EMT) and colorectal cancer(CRC) by detecting the expression of TGF-β, Smad-3 and Smad-7 in serum from CRC patients. Methods We detected the expression levels of TGF-β1 , Smad-3 and Smad-7 by ELISA method. Results The expression levels of TGF-β1 , Smad-3 were significantly higher and the Smad-7 was significantly lower in CRC patients than those in control group(P<0. 01). The expression levels of TGF-β1 and Smad-3 were signifi-cantly higher in Dukes D than those in Dukes B,C. The Smad-7 was significantly lower in Dukes D than that in Dukes B,C(P<0. 01). Conclusion TGF-β1 and Smad-3 and Smad-7 may play an important role in the develop-ment of CRC.

Objective To evaluate the clinical efficacy of interventional therapy by direct puncture for the treatment of avascular necrosis of the femoral head. Methods 24 experimental rabbits were undertaken and divided randomly into 3 groups, B, C groups as the models of hormone induced necrosis of femoral head, and A group as the control one. B group was followed up later with interventional therapy by direct puncture. Thromolytic agents, drugs improving microcirculation, and repairing bone tissue were infused directly into the femoral head; and then microscopic observations were conducted to study the pathology. Results The results showed: in group B, the number of vacant bony lacunae in femoral head, the average diameter of fat cells in marrow cavity and the number of blood vessel in subchondral areas showed significant improvement, approaching group A. Conclusions Treatment of femoral head necrosis through interventional method of direct puncture has commenced a solid foundation for clinical therapy.

Objective To obtain the recombinant IgE-dependent histamine-releasing factors of Schistosoma japonicum and Clonorchis sinensis (rSjHRF and rCsHRF) and to study the effect of recombinant HRFs to induce histamine release from sensitized rat mast cells. Methods The complete coding regions of SjHRF and CsHRF were cloned separately, and the recombinant plasmids were respectively transformed and expressed in BL21 cells. The soluble recom-binant rSjHRF and rCsHRF were purified. Aliquots of the mast cells obtained from the lungs of OVA-immunized rats were separately incubated with rSjHRF and rCsHRF and the released histamine was measured by the OPT spectrofluorometric procedure. The dose-dependent curves and the kinetics of histamine release induced by rSjHRF and rCsHRF were prepared. Results The recombinant plasmids pET-30-rSjHRF and pET-30-rCsHRF were constructed successfully and the purified soluble recombinant proteins rSjHRF and rCsHRF were obtained by affinity chromatography. rSjHRF and rCsHRF induced histamine release from sensitized mast cells in a dose-dependent manner. At the concentration of 150 mg/L, the average rate of histamine release from sensitized mast cells induced by rSjHRF and rCsHRF were 49.78% and 32.63%, respectively. Histamine release increased with prolonged reaction time and the maximal release occurred at 35min. Conclusion The recombinant parasite-originated IgE-dependent HRFs show an effect of inducing histamine release from sensitized mast cells, suggesting that this protein would play a role in type I hypersensitivity in hosts with parasitic infections.

BACKGROUND:It is accepted that the best method for spermatogonial stem cells separation is using artificial cryptorchism model combined with surface makers.OBJECTIVE:To explore the feasibility of separation spermatogonial stem cells with ?6-integrin and c-kit as specific surface markers.DESIGN,TIME AND SETTING:The randomized control experiment was performed at the Renmin Hospital of Wuhan University from May to December 2006.MATERIALS:Forty adult,white Kunming mice with 6 weeks old were randomly divided into cryptorchidism and control groups,with 20 animals in each group.METHODS:Artificial cryptorchidism model was prepared by made an incision at the median of abdomen,and testis was pulled into abdominal cavity,which was fixed at the each side of lateral abdominal wall.There was no treatment in the control group.The single cell suspension of seminiferous epithelium was obtained by traditional two step enzyme digestion at 2-3 months after operation.FITC-conjugated anti-?6-intergrin antibody and PE-conjugated anti-c-kit antibodies were added.Then the cells with low side scatter light-scattering properties were sorted and positively stained for ?6-intergrin and negative c-kit expression.Meanwhile,the viability of the isolated cells was assessed by trypan blue staining.MAIN OUTCOME MEASURES:The morphological changes of cryptorchidism,and the sorting results of spermatogonial stem cells.RESULTS:Cell distribution in seminiferous tubule was disorder with reduced numbers.The layer and lumens were disappeared,and cell division phase could be seen in the center of tubules.Compared to the control group,the testicular cells in the cryptorchidism group were increased in the side scatterlow,Forward scatterhi areas,with figure left-upward displacement.The distribution of ?6-integrin+ and c-kit cells were deviated each other,it named that most ?6-integrin+ cell were not spermatogonial stem cells,so do the c-kit-cells.Only 2.8% of testicular cells exhibited side scatterlow,?6-integrin+,and c-kit-,which were spermatogonial stem cells in the cryptorchidism group.And trypan blue staining showed that over 95% of them were viable.CONCLUSION:Using the two surface markers to sort spermatogonial stem cells can advance the purity of the spermatogonial stem cells in cell suspension,but the specificity is insufficient.

Melanin is a kind of biopolymer, composed of a complex quinine/indole-quinone derived mixture which is produced in melanocytes from tyrosine. More than 100 genes related to melanin deposition have been found in mouse. Slc24a5 (solute carrier family 24, member 5) is a novel gene first cloned in zebrafish. And it has been confirmed that the Slc24a5 is involved in controlling the melanin deposition in zebrafish. Here the full length of the chicken slc24a5 mRNA sequence, its expression profile and a discussion of its relationship to melanin deposition in chicken were reported. Chicken Slc254a5 has a CDS of 1 269bp, predicting a protein of 423 amino acids which is about 80 amino acids shorter than in human and mouse. It has 9 exons and 8 introns and spans more than 1lkb of genome sequence. The quantitative real-time PCR confirmed that the chicken Slc24a5 was highly expressed in the eye of White Leghorn and Beijing Fatty Chickens, and in the eye, skin and muscle of Silky. The relative expression in Silky eye is more than two times that of White Leghorn. The relative expression in Silky skin is about 70 times that of White Leghorn and about 15 times that of Beijing Fatty Chickens. The relative expression in Silky muscle is about 15 times that of White Leghorn and about 3 times that of Beijing Fatty Chickens. And the expression result is in accordance with the melanocyte and melanin deposition in these tissues.

BACKGROUND: Phenylketonuria is caused by gene mutation of phenylalanine hydroxylasel (PAH), which is mainly induced by permutation, short segments and insertion of base.OBJECTIVE: To evaluate the gene mutation of phenylalanine hydroxylasel in phenylketonuria in Hui nationality.DESIGN: Open study.SETTING: Urumqi General Hospital of Lanzhou Military Area Command of Chinese PLA; Capital Pediatrics Institute.PARTICIPANTS: A boy of Hui nationality in China and aged 3.1 years was selected in this study. The boy had intellect hysteresis in his one year and received medical treatment in his three years, while he was diagnosed as cerebral paralysis. After repeatedly inefficient treatment, he was hospitalized in our hospital on December 13, 2004. Iron sesquichloride in urine was strongly positive and concentration of serum phenylalanine was 1 680 μmol/L; therefore, he was diagnosed as the typical phenylketonuria.METHODS: 5 mL venous blood was selected from the boy and his parents, respectively, and anticoagulated with EDTA-Na2. DNA in gene group was extracted by using typical phenol/chloroform method. In addition, polymerase chain reaction (PCR) primer sequence of extron 7, 6, 11, 3, 12 and 5 of PAH gene was designed based on references. And then, PCR products were detected with 2% agarose gel electrophoresis. 5 μL PCR products were mixed with the same volume of degenerated buffer solution, degenerated at 97 ℃ for 5 minutes, put in iced bath and performed with 80 g/Lnon-degenerated polyacrylamide gel electrophoresis. After that, the products were dealt with sliver staining routinely, and single strand DNA banding patterns were analyzed and recorded. ABI377 automatic sequenator (PE Company) was used to detect PCR sequence and purify PCR product in Shanghai Boya Biotechnology Company.MAIN OUTCOME MEASURES: Iron sesquichloride in urine, concentration of serum phenylalanine and mutant gene types of phenylalanine hydroxylase.RESULTS: Extron 7, 6, 11, 3, 12 and 5 of PAH gene were analyzed in the boy and his parents. The results demonstrated that SSCP electrophoresis in extron 6 was different from that in the normal control group. Site of electrophoresis strip of his father was coincident with that of his mother, but different from that of the boy. Sequencing results indicated that point mutation (cytosine replaced by thymine), which was a R176X mutant heterozygote, occurred at the 526th site of cDNA of phenylalanine hydroxylase gene in his parents; however, two chromosomes of the boy had mutation at the same site, which was R176X mutant homozygote.CONCLUSION: Mutation of R176X homozygote of phenylketonurea is firstly reported in Hui nationality in China.

Objective To investigate whether chemokine CC motif 2 (CCL2) is involved in the high residual platelet response,and the mechanism of CCL2 being involved in the regulation of platelets.Methods Forty patients with ST elevation myocardial infarction (STEMI) were admitted.P2Y12 reaction unit (PRU) was detected by VerifyNow.Forty patients were divided into high platelet reactivity group (high reactivity group,n=24) and normal platelet reactivity group (normal reactivity group,n=16) according to the results of PRU detection.Plasma CCL2 concentration of the STEMI patients was examined by ELISA.The expressions of CCL2 and CCR2 in the platelets were detected by Western blotting.After CCL2 stimulation,the kinases of which phosphorylation was changed in the platelets were screened by ARY003B protein chips.The phosphorylation of p38MAPK and HSP27 in the platelets was tested by Western blotting after CCL2 stimulation in the presence or absence of CCR2 antagonist (RS 102895) or p38MAPK signal pathway inhibitor (SB 203580).Results The plasma CCL2 concentration of high reactivity group was markedly higher than that of normal reactivity group.Moreover,compared with normal reactivity group,the expressions of CCL2 and CCR2 in the platelets of high reactivity group significantly increased.After the platelets were stimulated by CCL2,the phosphorylation of p38α and HSP27 enhanced in the platelets by protein chips screening.When RS 102895 or SB 203580 was treated before CCL2 stimulation,the phosphorylation of p38MAPK and HSP27 decreased.Conclusions CCL2 participates in high residual platelet response in an autocrine/paracrine way.CCL2/CCR2 might affect the function ofplatelets through p38MAPKHSP27 signal pathway.

Objective To investigate expression of 4-1BB and lymphocyte subsets in peripheral blood of children with acute infectious lymphocytosis. Methods Flow cytometry (FCM) was applied to detect the expression of 4-1BB and lymphocyte subsets in peripheral blood of 15 cases of acute infectious lymphocytosis and 20 cases of acute upper respiratory infection, and 20 healthy children. Results The expression of 4-1BB and CD3+, CD4+and CD8+lymphocytes were higher in acute infectious lymphocytosis group than those in acute upper respiratory infection group and healthy control group (P<0.05). There was no sig-niifcant difference of CD19+CD23+lymphocytes among three groups (P>0.05). A positive correlation was found between 4-1BB expression and CD3+ lymphocytes expression in acute infectious lymphocytosis group (r=0.73, P<0.05). Conclusions The abnormal expression of 4-1BB may play a pathological role in the development of acute infectious lymphocytosis. T cells in chil-dren with acute infectious lymphocytosis may not function to activate B cells.

OBJECTIVE To study the mechanism of drug-resistance of multi-resistant Pseudomonas aeruginosa,and provide the guideline for treatment and control of P.aeruginosa infection in hospital.METHODS Fifty strains of multi-resistant P.aeruginosa were selected with K-B susceptibility method.The three-dimensional method was taken to differentiate the various beta-lactamases.The relative drug-resistance gene was detected by polymerase chain reaction(PCR).RESULTS Among 50 strains of multi-resistant P.aeruginosa,there were 2 strains(4%)producing ESBLs,20 strains(40%)producing AmpC beta-lactamases,and 11 strains(22%) producing ESBLs and AmpC beta-lactamases at the same time.There were 8 positive genes in the detected drug-resistance gene,the most common sources of gene were CTX(56%),OprD(60%) and aac(6′)-Ⅱ(60%),respectively.CONCLUSIONS The main beta-lactamases are AmpC beta-lactamases and the main genotype is CTX in the multi-resistant P.aeruginosa cultured in our area.The main course of imipenem-resistance was deletion of outer membrane proteins,and the aminoglycoside modifying enzyme gene and disinfectant-resistance gene in multi-resistant P.aeruginosa are acquired.In order to reduce the drug-resistance strains and control the infection of P.aeruginosa,antibiotics should be used reasonably according to drug susceptibility testing clinically.

In this study ECG signal of unstrained rat was recorded by telemetry device, and heart rate variability (HRV) was analyzed in order to evaluate 24h autonomic nervous activity. The results demonstrated an obvious circadian rhythm in the autonomic nervous activity: sympathetic activity being dominant during wake phase, and parasympathetic activity, dominant during sleep phase. The ratio of the low frequency to high frequency components in HRV power spectrum (LF/HF) fluctuates with the change in the sleep stages. It is concluded that 24h HRV analyses may reveal plentiful information about the behavior of autonomic nervous system and thus facilitate the investigation of its regulating role in physiological and pathological processes.
Animals ; Autonomic Nervous System ; physiology ; Circadian Rhythm ; Electrocardiography ; Heart Rate ; physiology ; Male ; Rats ; Rats, Sprague-Dawley ; Telemetry