Objective To clone and express the cDNA encoding Schistosoma japonicum tropomyosin. Methods The cDNA was amplified by reverse transcription polymerase chain reaction (RT PCR). The PCR products were ligated with pGEM T vectors and then for transformations. After characterization of white clones by agarose gel electrophoresis, endonucleases digestion and PCR, some recombinant plasmids with inserts were used for sequencing. Then the gene was subcloned into prokaryotic expression vector pQE30 and expression was induced by IPTG. Results The PCR products was 823 bp judged by agarose gel electrophoresis and sequencing. A cDNA encoding S japonicum tropomyosin, except for 14 amino acids at the amino terminus and 2 at the carboxyl terminus, has been constructed and cloned successfully. The colony, designated pGSjcTM12, was sequenced and shown to be 91 1% identical at the nuclei acid level and 98.1% identical in deduced amino acid sequence to that of S mansoni tropomyosin. The gene was subcloned into pQE30 and an expressed protein of about 32 kDa was obtained.Conclusion The cloning and expression of the gene encoding S japonicum tropomyosin had been successfully made.

Objective To investigate the difference of differentiation from rat bone marrow mesenchymal stem cells(MSCs) from normal group and hepatic fibrosis model group into hepatocyte-like cells.Methods Wistar rats were randomly divided into two groups: normal group and hepatic fibrosis model group.The liver fibrosis was induced by CCL4.MSCs were isolated by combining gradient density centrifugation with plastic adherence.Pure MSCs were obtained by cultivation and passage.The cells then treated with HGF and FGF-4.Levels of AFP and albumin from supernatant were determined on day 15,21 and 27.On day 27,cells of induced and non-induced were collected,glycogen store of hepatocytes and the expressions of CK-18 and CK-19 were detected.Results The level of AFP in induced MSCs was higher on day 15,21,27,and reached the peak on day 21;there was no significant difference between induced and non-induced MSCs in albumin levels on day 15,but on day 21,27,compared with the non-induced MSCs,the albumin level in the induced MSCs was higher and reaches its peak on day 27;glycogen storage of induced MSCs was measured on day 27 as compared with non-induced MSCs;the induced MSCs expressed CK-18 and CK-19 while the non-induced MSCs did not.Compared by the levels of AFP and albumin,there was no significant difference in differentiation effect of MSCs between the normal group and hepatic fibrosis model group.Conclusion Rat MSCs of hepatic fibrosis model group could differentiate into hepatocyte-like cells with hepatic phenotype and biological function in the presence HGF and FGF-4.

BACKGROUND: The implantation technique of bone marrow mesenchymal stem cells (MSCs) is of important significance for repair of brain injury. However, its action pathway still needs to be investigated.OBJECTIVE: To observe the changes of Bcl-2 and Bax expressions in the injured regions of cerebral cortex and hippocampus of rats with cerebral ischemia in which MSCs were implanted, and to analyze the action mechanism of intracranial implantation of MSCs inhibiting neuronal apoptosis.DESIGN: A randomized controlled experiment.SETTING: Institute for Cerebrovascular Disease, the Affiliated Hospital of Qingdao University Medical College.MATERIALS: Twenty-four male adult Wistar rats, weighing 180 to 240 g, were provided by the Experimental Animal Center of Shandong University. The involved rats were randomized into 3 groups with 8 in each: control group, injury group and implantation group. 5-bromodeoxyuridine (BrdU, Sigma Company), TUNEL kit, Bcl-2, Bax antibody kit were purchased from Beijing Zhongshan Bioengineering Company.METHODS: This study was carried out in the Institute for Cerebrovascular Disease, the Affiliated Hospital of Qingdao University Medical College between April 2005 and September 2006. Rats in the injury group and implantation group were developed into rat models of cerebral ischemia by suture of external carotid artery. Seven days later, the successful rat models in the implantation group were injected in the cerebral cortex and striatum with 2×1012 L-1 MSCs suspension primarily cultured in vitro. The processing of the experimental animals corresponded to the requests of Animal Ethics.MAIN OUTCOME MEASURES: Neuronal apoptosis and Bcl-2 and Bax expressions in the injured regions of cerebral cortex and hippocampus were detected by TUNEL staining and immunohistochemical method on the 7 and 14 days after successful modeling, separately.RESULTS: ①Neuronal apoptosis: On the 7th day after successful modeling, apoptotic cells were not found in the control group, and apoptotic cells in the implantation group were significantly fewer than those in the injury group (P ＜0.01). ② Bcl-2 and Bax expressions: On the 14th day after successful modeling, Bcl-2-positive neuronal expression in the cerebral cortex and hippocampus of rats in the injury group was significantly weaker than that in the control group and implantation group (P ＜ 0.01). Bax-positive expression in the cerebral cortex and hippocampus of rats in the injury group was significantly stronger than that in the control group and implantation group (P ＜ 0.01).CONCLUSION: MSCs can promote Bcl-2 expression and inhibit Bax expression of rats with cerebral ischemia injury,and accordingly neuronal apoptosis will be reduced.

Objective To study the polymorphism of human platelet antigen HPA-1 to HPA-5,and HPA-15 system in Qingdao Han population.Methods A total of 918 samples from regular voluntary platelet donors in Qingdao were genotyped for HPA-1 to-5 and HPA-15 by PCR-SSP.Results The gene frequencies of HPA-1a,-1b;HPA-2a,-2b;HPA-3a,-3b;HPA-4a,-4b;HPA-5a,-5b;HPA-15a,-15b were 0.9940,0.0060;0.9319,0.0681;0.5822,0.4178;0.9897,0.0104;0.9804,0.0196;0.4913,0.5087,respectively.Both a and b alleles were found in each of the 6 HPA systems,and a/a homozygosity was more common in HPA-1,-2,-4 and-5 systems.The HPA genotype frequencies followed Hardy-Weinberg principle.HPA-1 frequency of Qingdao people was significantly different from that of North China(P

ObjectiveTo construct and identify a DNA vaccine of Schistosoma japonicum (Chinese strain) thioredoxin. MethodsAccording to a cDNA sequence of S.japonicum (Philippine strain) thioredoxin, a couple of primers was designed with the BamH I restriction endonuclease site introduced in forward primer with ATG as start condon and EcoR I in reverse primer with TCA as termination codon. The gene encoding S.japonicum (Chinese strain) thioredoxin (SjcTrx) was amplified by RT-PCR using total RNA of schistosome as the template. The PCR products and the pcDNA3 plasmids were digested by restriction endonucleases BamH I and EcoR I. The target DNA fragment were purified and cloned properly into the eukaryotic expression vector of pcDNA3. The recombinant plasmids were then transformed into competent E.coli JM109 and identified by endonucleases digestion, PCR, agarose gel electrophoresis and sequencing. ResultsThe RT-PCR product was around 334 bp judged by agarose gel electrophoresis. The same fragments were obtained by restriction enzyme digestion from the recombinant plasmid and PCR with the plasmid DNA as a template. The recombinant plasmid, designated pcDNA3-SjcTrx, was sequenced and shown to be 97% and 43% identical in deduced amino acid sequences to that of S.japonicum (Philippine strain) and S.mansoni thioredoxin, respectively. ConclusionThe S.japonicum thioredoxin DNA vaccine had been constructed successfully, and further studies will be made in different animal models for its immunogenicity.

National traditional Chinese medicine (TCM) master professor Ban Xiuwen had rich experiences on treatment of gynecological diseases with unique insights on diagnosis and treatment for disorders of menstruation, leukorrhea, pregnancy and delivery. FemaleYin-Wei, which was a gynecological disorder, was recorded in the book ofHuang-Di Nei-Jing. However, due to different reasons, TCM study on femaleYin-Wei was far less than the study on male impotence. ProfessorBan Xiuwen had specific considerations for this disease on the book of Treatment of Miscellaneous Gynecological Diseases. In his understanding, femaleYin-Wei was due to four aspects, which were the innate deficiency, yang deficiency and coldness of the uterus; seven emotion damage, liver-qi soothing disorder; spleen-stomach deficiency,qi-blood deficiency; and phlegm-dampness stasis,qi stagnation. Through the analysis and study on case records of femaleYin-Wei treatment by national TCM master professorBan Xiuwen, this article briefly elaborated professorBan Xiuwen’s understandings on femaleYin-Wei treatment from etiology, pathogenesis, treatment according to syndrome differentiation, and clinical experiences. Two femaleYin-Wei cases treated by professorBan Xiuwen were given as examples in order to analyze the etiology, pathogenesis, treatment according to syndrome differentiation and treatment experiences in details. The therapeutic effects of both cases were obvious, which were also important study documents.

The iodine concentration, MIT/DIT and T3/T4 ratios and peroxidase activity in thyroid of the rats fed on endemic goiter region diet of county Chefong in the Inner Mongolia Autonomous Region and control rats injected with 30% iodine oil 0.2 ml besides the same diet were investigated.The result demonstrated that the iodine concentration of thyroid was decreased with increasing of thyroid weight, and the MIT/DIT ,T3/T4 ratios in thyroid and thyroid peroxidase activity of the rats fed on endemic region diet were higher than that of control.These metabolic patterns showed that the rats fed on endemic region diet were in iodine deficient state and consistent with those rats fed on artificial low iodine diet.

Objective To understand and identify the molecules related to the natural resistance to Schistosoma japonicum infection in Mirotus fortis. Methods Sera from Mirotus fortis without schistosome infection were collected. The S.japonicum adult worm cDNA library was immunologically screened with the sera. The positive recombinants were identified, cloned, sequenced and analysed with software and internet. Results Seven genes encoding antigens relevant to sera antibodies in Mirotus fortis were cloned and sequenced. These antigens included glyceraldehyde 3-phosphate dehydrogenase (GAPDH), serine protease inhibitors(SERPIN), 70 kDa heat shock protein(HSP70), 22\^6 kDa membrane-associated antigen, paramyosin (Sj97), cytochrome C and cathepsin B. Conclusion Many protein molecules might have been involved in natural resistance to \{S.japonicum\} infection in Mirotus fortis. The above 7 kinds of molecules may be identified as new candidates of vaccine against \{S.japonicum\} infection.

Objective To investigate the therapeutic effect of ambroxol in bleomycin-induced pulmonary fibrosis of rats.Methods Sixty Wistar rats were divided into three groups:The normal control group(group N),the model group treated with bleomycinA5(group B)and the group treated with ambroxol(group A).Pulmonary fibrosis was induced by intratracheal instillation of bleomycin.Then the rats received daily ambroxol 35mg/kg(group A),or normal saline(group B).Five rats in each group were killed on 3.7.14 and 28 days after intratracheal instillation.Histological changes of the lungs were evaluated by HE stain and Masson's trichrome stain.The levels of tumor necrosis factor-?(TNF-?)in lung homogenates were measured by radioimmunoassay.Results Pulmonary fibrosis of Group A was significantly improved as compared with that of Group B(P

Objective To summarize the clinical, electrophysiological, pathological characteristics of chronic inflammatory demyelinating polyradiculoneuropathy (CIDP). Methods From Jan 1990 to Dec 2003, 32 patients of CIDP were admitted by our hospital, 14 male and 18 female. The age range of onset of illness was 13 to 74 years old (averaging 42.56) with the prime period of 40 to 50. Before hospitalization, the course was from 2 months to 5 years. Among all patients, there were 8 cases of a relapsing course and 24 cases of a chronic progressive course. Twenty-four patients were treated with corticosteroids, while 7 received immuglobin (IG) and corticosteroids. The clinical data before and after the treatment was studied retrospectively. Results Most of the patients had subacute or chronic onsets. The common initial symptoms are numbness, paresthesia, and extremity weakness. Diplopia, decreased visual acuity, dysarthria, and dysphagia could also be found initially. It was usually a symmetric sensorimotor neuropathy with either a relapsing course or a chronic progressive course. It could be accompanied with autonomic dysfunction and cranial nerve involvement. Electromyogram demonstrated that the motive and sensory nerve conduction velocities were slow. The sural nerve biopsy showed demyelination and remyelination. `IG and corticosteroids were both effective. Conclusion CIDP might result in widespread peripheral nerve damages, in which autonomic dysfunction and cranial nerve involvement were common. The dominant electrophysiological changes showed peripheral nerve demyelination accompanied by axon degeneration. The sural nerve biopsy played an important role of diagnosis. The treatment with IG and corticosteroids was a most effective way for CIDP nowadays.