Objective To clone and express a membrane protein(Tetraspanin 2) gene of Schistosoma japonicum (SjTsp2). Methods A pair of primers was designed to amplify the SjTsp2 gene which was subcloned into prokaryotic plasmid pET28a(+). The recombinant plasmid was transformed into E. coli BL21(D3) and followed by expression of the protein induced by IPTG. The protein was purified by affinity chromatography and used to immunize BALB/c mice. Dilution of antibody against SjTsp2 was determined by ELISA. The protein was also identified by Western blotting. Results Big loop of SjTsp2-A, 228 bp, was amplified in vitro by PCR. Its deduced amino sequence shared 52% similarity with SmTsp2. The soluble recombinant SjTsp2-A was expressed in the experiment and high dilution antibody against the recombinant (1 ∶ 32 000 in maximum) was produced in immunized mice. SjTsp2-A reacted positively with sera of acute and chronic schistosomiasis patients but not with sera from healthy persons by Western blotting. Conclusion SjTsp2 has been expressed and shows certain antigenicity.

Tetraspanin 2-A(SjTsp2-A) gene was amplified by PCR.pcDNA3.1(+)/SjTsp2-A recombinant plasmids were constructed and transformed into E.coli DH5?.Twenty four BALB/c mice were randomly divided into pcDNA3.1(+) /SjTsp2-A group(A),pcDNA3.1(+)/SjGST group(B) and pcDNA3.1(+) group(C).Each mouse was injected through musculus quadriceps femoris by three times(two weeks interval) respectively with 100 ?g pcDNA3.1(+)/SjTsp2-A,pcDNA3.1(+)/SjGST,or pcDNA3.1(+).At two weeks after the final inoculation,mice were each challenged by 40?2 cercariae of S.japonicum.Forty-five days after infection,all mice were sacrificed,the number of worms collected and eggs in liver tisssue was counted.Anti-pcDNA3.1(+)/SjTsp2-A antibody was detected by ELISA and protein expression in quadriceps muscle by immunohistochemical staining.The worm reduction rate(44.4%) and egg reduction rate(28.4%) of group A was higher than those of group B and C(P

Objective By observing the effect of Xiaoyuhuatan decoction on insulin resistance in rar with nonalchoholic fatty liver,the author made a reasearch into this herb'S machanism on cambating on nonalchoholic fatty liver.Methods The insulin resistance rat model was fed by high fat forage and Dongbaogantai control group was set up for the sake of comparion.We determined the contents of total cholesterol(TC),triglyceride(TG),free fatty acid(FFA),fasting plasma glucose(FBG),fasting serum insulin(INS)in serum and the contents of TC,TG of liver tissue homogenate of each group,and the degree of hepatocytic steatosis.Results Xiaoyuhuatan decoction could obviously decrease the index of insulin resistance and improve insulin insensitive index.Lipid in blood serum and hepatic tissues were significantly decreased,liver fat cell denaturation was improved.Conclusion The thrapentic eftect of Xiaoyuhuatan Decoction on fatty liver is probably contribute tO this herb's mechanism of improving insulin resistance and increasing insulin insensitive.

AIM: To observe the effect of Xiaoyu Huatan Decoction on the nonalchoholic fatty liver tissue peroxisome preliferator-activated receptor?(PPAR?) expression. METHODS: The nonalchoholic fatty liver disease rat model was fed by high fat forage and the rats were divided into five groups: normal control group,model control group,high-dose of Xiaoyu Huatan Decoction group,low-dose of Xiaoyu Huatan Decoction group,Dongbao Gantai control group.Total RNA of liver was extracted,and the expression of PPAR?mRNA was analyzed by semi-quantitative RT-PCR method.We determined the contents of total cholesterol(TC),triglyceride(TG),free ratty acid(FFA) in serum and the TC,TG in liver tissue homogenate of each group,and the degree of hepatocytic steatosis. RESULTS: Expression of liver tissue PPAR? mRNA in the model group decreased significantly,lipid in blood serum and hepatic tissues increased significantly,liver fat cell greatly denaturalized.After the intervention of medicine,expression of liver tissue PPAR? mRNA of each treatment group increased significantly,lipid in blood serum and hepatic tissues decreased significantly,liver fat cell denaturation was improved. CONCLUSION: Xiaoyu Huatan Decoction can increase the expression of liver tissue PPAR?mRNA of rats,It is likely to be one of the important mechanism for treating fatty liver.

Objective To observe the effect of Xiaoyu Huatan Decoction (XHD) on hepatic apoptosis and apoptosis- related gene Bcl-2,Bax expression in nonalcoholic fatty liver disease(NAFLD)rats,and to explore its therapeutic ef- fect for NAFLD.Methods The rat model of NAFLD was induced by feeding high fat forage.Dongbao Gantai was used as the positive control drug.Flow cytometry(FCM)was used to detect the hepatic apoptosis and expression of Bcl-2 and Bax,and optical microscope was used to observe the degree of hepatic steatosis.Results Moderate or serious steatosis of liver cells was tound under optical microscope,and there existed a few hepatic necrosis.The apoptotic rate and FI expres- sion of Bax in the model group were significantly increased (P

Objective:To analyze the clinical features, imaging findings and gene test of patients with type Ⅱ Alexander disease.Methods:All the clinical data of three cases with type Ⅱ Alexander disease from August 2018 to June 2020 in the Department of Neurology, Qilu Hospital of Shandong University (Qingdao) and Qilu Hospital of Shandong University were collected, and their clinical and imaging findings were analyzed retrospectively.Results:All the three patients were middle aged and old men with a chronic progressive course, beginning with weakness of one or both lower limbs, followed by dizziness, dysarthria, dysphagia, sphincteral disturbances, constipation and orthostatic hypotension. Three patients all experienced misdiagnosis (hydrarthrosis, cerebral vascular disease, alcoholism, respectively) at early stage of the disease. Cranial magnetic resonance imaging (MRI) showed mild supratentorial periventricular leukodystrophy, which was not specific. Sagittal cranial MRI demonstrated medulla oblongata and upper cervical cord atrophy called “tadpole atrophy”, which had high suggestive value. The results of gene analysis showed heterozygous mutation of glial fibrillary acidic protein gene, which had been reported as pathogenic gene; c.1091C>T (p.A364V) in exon 6, c.722C>T (p.R258C) in exon 4 and c.197G>A (p.R66Q) in exon 1, respectively.Conclusions:Type Ⅱ Alexander disease is an autosomal dominant disease, most with point mutations, rarely with deletion mutations. Type Ⅱ Alexander disease should be suspected when a patient had signs of lower brainstem involvement such as dizziness, ataxia, pyramidal sign, autonomic dysfunction, especially when cranial MRI showed mild supratentorial leukodystrophy, and medulla oblongata and upper cervical cord atrophy.

OBJECTIVETo identify the endophytic fungal strain PR35 separated from Paeonia delavayi and study chemical constituents of its secondary metabolites.

METHODThe fungal strain PR35 was identified by morphological observation and ITS rDNA sequence analysis. Various chromatographic methods were adopted to separate and purify its secondary metabolites, and their structures were identified by physiochemical properties and spectral data

RESULTThe fungal strain PR35 was identified as Trichoderma longibrachiatum. Five compounds were separated from fermentation products of fungal strain PR35 and identified as 1-(2,6-dihydroxyphenyl)-3-hydroxybutan-1-one (1), 1-(2,6-dihydroxypheny) propan-1-one (2), 1-(2,4,6-trihydroxyphenyl) butan-1-one (3), 4-methoxy-1-naphthol (4), and cerevisterol (5). Among them, compounds 1-3 showed notable antifungal activities against Botrytis cinerea, Fusarium avenaceum and Hormodendrum compactum.

CONCLUSIONThe endophytic fungus T. longibrachiatum was separated from the plant P. delavayi for the first time. Five compounds were first separated from endophytic fungus of P. delavayi. Among them, compound 4 was separated from microbial fermentation products for the first time.

DNA, Fungal ; genetics ; DNA, Intergenic ; genetics ; Endophytes ; classification ; genetics ; isolation & purification ; metabolism ; Paeonia ; microbiology ; Phylogeny ; Trichoderma ; classification ; genetics ; isolation & purification ; metabolism

Objective:To study the characteristics of clinical, laboratory, imaging, genetic and differential diagnosis of McLeod syndrome.Methods:The clinical characteristics of 2 cases of McLeod syndrome confirmed by gene detection in Qilu Hospital (Qingdao) on June 27, 2018 and in Qilu Hospital of Shandong University on September 11, 2019 were analyzed retrospectively. And the characteristics of patients of McLeod syndrome reported in China were analyzed in combination with literature review.Results:Both of the 2 patients were adult male, aged 57 and 61 years, respectively, with a slowly progressive course, beginning with gradually involuntary movement of trunk and extremities, involving involuntary biting of the tongue and dysphagia. Two patients had mild cognitive impairment; one patient had emotional agitation. Imaging study showed atrophy of caput nuclei caudate. Neuroelectrophysiological examination of case 1 showed sensory axon neuropathy in both upper limbs with severe damage to the left ulnar nerve. Creatine kinase (CK) was mildly elevated in 2 patients. The peripheral blood smear of 1 patient showed increased acanthocytes, accounting for 13%, the other patient showed no increased acanthocyte. McLeod syndrome related gene was tested in the 2 patients, case 1 with deletion mutation of exon 2 of XK gene, and case 2 with hemizygotic mutation of XK gene c.898delC p.L300 *. Conclusions:The clinical manifestations of McLeod syndrome are various and the differential diagnosis is crucial. For elderly male with cephalic facial chorea, elevated CK level and neuromuscular diseases, the possibility of McLeod syndrome should be screened.

OBJECTIVETo investigate the effect of the lentiviral vector mediated CXCR4 overexpressed mesenchymal stem cell (MSCs) on graft-versus-host disease (GVHD).

METHODSLentiviral vector containing CXCR4 was constructed. CXCR4 overexpressed MSC by lentiviral vector mediated were assessed. A major histocompatibility complex (MHC)-mismatched mouse model of bone marrow transplantation (BMT) from C57BL/6 donors to BALB/c recipients was constructed. Mice were divided into five groups: total body irradiation (TBI) group, mice received irradiation only; BMT group, mice were transplanted with bone marrow (BM) after TBI; GVHD group, mice were transplanted with BM and splencytes after TBI; CXCR4-MSC group, mice were transplanted with CXCR4-MSC, BM and splencytes after TBI; EGFP-MSC group, mice were transplanted with EGFP-MSC, BM and splencytes after TBI. The survival, body weight and clinical score of GVHD in transplanted mice were monitored. Liver, intestine and skin from mice in each group were obtained for histological examination. Plasma concentrations of inflammation factors such as interleukin (IL)-2, IFN-γ and TNF-α were also determined using a cytometric bead array cytokine kit.

RESULTSAll mice in TBI group died within 14 days, while all of BMT group survived. The mean survival times for GVHD, EGFP-MSC and CXCR4-MSC groups were (17.0 ± 2.3) d, (21.7 ± 4.8) d and (30.1 ± 9.1) d, respectively. Treatment with CXCR4 over-expressing MSCs could decrease the mortality rate. All mice in each group developed clinical signs such as hunched posture, dull fur, diarrhea and weight loss. Meanwhile, histopathological findings in target organs were confirmed the presence of GVHD. While, clinical GVHD scores and histopathological scores in CXCR4-MSC group were significantly lower than that of GVHD group. Moreover, compared with control groups, the plasma IL-2, IFN-γ and TNF-α level in recipients infused with CXCR4-MSC were significantly decreased (P<0.05).

CONCLUSIONThe results revealed that CXCR4- transduced MSCs could effectively control the occurrence of mouse GVHD following allogeneic BM transplantation.

Animals ; Bone Marrow Transplantation ; Cytokines ; Genetic Vectors ; genetics ; Graft vs Host Disease ; Lentivirus ; genetics ; Mesenchymal Stromal Cells ; immunology ; Mice ; Receptors, CXCR4 ; genetics ; immunology ; Transplantation, Homologous