Objective To make molecular diagnosis for puerile spinal muscular atrophy(SMA).Methods Genomic DNA was extracted directly from the blood of both the case group(10 children with SMA) and the control group(including 19 parents of SMA patients and 20 healthy individuals).Two methods,polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) and allele-specific PCR,were used to analyze exon 7 of SMN gene from genomic DNA,and consequent electrophoresis of PCR products on agarose gel was performed.Results Genotyping results obtained by both methods were in complete agreement for all of the samples analyzed.In conventional PCR-RFLP,part of the PCR products(189bp) from genomic DNA of all 39 members in the control group remained intact after digestion with Dra I,while the PCR products from genomic DNA of all 10 SMA children in the case group was completely digested by Dra I.In allele-specific PCR,exon 7 of both SMN1 and SMN2 could be seen when genomic DNA of all 39 members in the control group was used,while only SMN2's exon 7 could be seen when genomic DNA of all 10 SMA children in the case group was used.Conclusion Homozygous deletion of SMN1 was present in all 10 SMA children in the case group,while homozygous deletion of SMN1 was not detected in all 39 members in the control group.The combination of PCR-RFLP and allele-specific PCR,both their results can be references for each other,offers efficient and accurate methodology for molecular diagnosis of SMA.

Hydrophidae, one of the precious traditional Chinese medicines, is generally drily preserved to prevent corruption, but it is hard to identify the species of Hydrophidae through the appearance because of the change due to the drying process. The identification through analysis on gene barcode, a new technique in species identification, can avoid the problem. The gene barcodes of the 6 species of Hydrophidae like Lapemis hardwickii were aquired through DNA extraction and gene sequencing. These barcodes were then in sequence alignment and test the identification efficency by BLAST. Our results revealed that the barcode sequences performed high identification efficiency, and had obvious difference between intra- and inter-species. These all indicated that Cyt b DNA barcoding can confirm the Hydrophidae identification.
Animals ; Base Sequence ; China ; Cytochromes b ; genetics ; DNA Barcoding, Taxonomic ; Elapidae ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Reptilian Proteins ; genetics ; Sequence Analysis, DNA

The effect of triptolide (TP) on the expression of receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) was explored in rat adjuvant induced arthritis (AA). AA was induced in Wistar rats. Arthritis rats were treated with TP and methotrexate (MTX) at the onset (day 9) of arthritis. On the peak of arthritis (day 24), the expression of RANKL and OPG protein in the joints and RANKL mRNA in peripheral blood mononuclear cells (PBMC) was detected. TNF-alpha and IL-1beta levels in peripheral blood were determined. Bone erosion scores were also evaluated. The results showed that bone erosion scores in TP and MTX groups were lower than in AA group (P < 0.01); The expression levels of RANKL in the synovium (P < 0.01) and bone (P < 0.05), and OPG level in synovium (P < 0.05) were lower in TP group than in AA group (P < 0.05). In TP group, the expression levels of RANKL mRNA and TNF-alpha, IL-1beta in PBMC were lower than in AA group (all P < 0.01). It was concluded that TP could inhibit rat adjuvant arthritis bone erosion by suppressing the expression of RANKL.

BACKGROUND: Common threewingnut root has the functions of anti-inflammation and immune inhibition, etc., and it has been used at present to treat various autoimmune diseases including rheumatoid arthritis.. Common threewingnut root has complex components, and triptolide is acknowledged as one of the important effective components of common threewingnut root.OBJECTIVE: To establish rat models of type Ⅱ collagen induced arthritis, and observe the effects of triptolide on the contents of interleukin-6,interleukin-10 and tumor necrosis factor alpha (TNF-α) in peripheral serum and synovial fluid.DESIGN: A randomized control animal experiment.SETTING: Department of Integrated Traditional and Western Medicine,Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiments were carried out in the Tongji Hospital from November 2004 to July 2005. Fifty healthy male Wistar rats of clean degree were purchased from the experimental animal center of Tongji Medical College, Huazhong University of Science and Technology [qualification number of animal [scxk(E)2004-2007]. Triptolide (nobatch number because of temporary production) was bought from Fujian Institute of Medical Sciences, and the purity was above 98.5%.METHODS: ① Ten of the 50 rats were randomly selected as the normal controls, and the others were made into models. Type Ⅱ collagen emulsion was injected intradermally at five points along the back and tail of the rats,0.05 mL for each point, and injected intradermally at two points after 15 days. The rats in the normal control group were treated with saline in the same way. The effects of the model establishment were evaluated according to the scoring standards of arthritis index at 30 days after the first immunity, and the rats scored 6 points or above were taken as successful models and enrolled in the experiments. Twenty successful rat models were randomly divided into arthritis model group (n=10) and triptolide treated group (n=10). ② Triptolide (100 mg/L)was dispensed into parenteral solution with propylene glycol (0.05 in volume fraction), and then intramuscularly injected into hindlimb of rats in the triptolide treated group (0.04 mL/100 g), once every three days for 30 days. The rats in the normal control group were given isovolume saline, and those in the arthritis model group were treated with isovolume propylene glycol (0.05 in volume fraction). ③ The materials were removed at 30 days after administration. The contents of interleukin-6, interleukin-10 and TNF-α in peripheral serum and synovial fluid were detected with enzyme-linked immunosorbant assay(ELISA).MAIN OUTCOME MEASURES: The effects of triptolide on contents of TNF-α, interleukin-6 and interleukin-10 in peripheral serum and synovial fluid were observed.RESULTS: Fifty male Wistar rats of clean degree were selected, 10 were used as normal controls, and 20 of the other 40 rats were successfully made isto models and enrolled in the analysis of results. ① The TNF-α contents in peripheral serum and synovial fluid were the highest in the arthritis model group, and obviously decreased after treatment of triptolide [(35.09±8.82), (15.35±3.56) ng/L; (44.17±8.94), (22.54±4.76) ng/L; P＜ 0.01], which were similar to those in the normal control group (P ＞ 0.05).② The contents of interleukin-6 in peripheral serum and synovial fluid were the highest in the arthritis model group, and were obviously decreased after treatment of triptolide [(76.58 ±6.81), (42.45 ±5.72) rig/L;(88.69±10.56), (48.67±5.97) ng/L; P ＜ 0.01], but did not recover to the levels in the normal control group (P ＜ 0.05). ③ The contents of interleukin-10 in peripheral serum and synovial fluid were the lowest in the arthritis model group, and obviously increased after treatment of triptolide[(17.53±2.07), (21.23±2.91) ng/L; (10.59±2.96), (14.74±1.85) ng/L; P＜ 0.01], which were similar to those in the normal control group (P ＞ 0.05).CONCLUSION: Triptolide can treat arthritis by modulating the contents of cytokines.

The effect of nifedipine on antipyrine ( AP ) disposition was evaluated in 10 patients with essential hypertension. The disposition were measured with high perfomance liquid chromatography before and after 5d nifedipine treatment. The results showed that APt_1/2 and CL_AP were 14.88?3.1h and 31.29?5.9 ml/min respectively before treatment; APt_1/2 and CL_AP were 14.37?3.1 h and 32?6 ml/min after treatment. There were no significant differences for both parameters between the pre-treatment and after-treatment. The results showed there was no effect of nifedipine on AP disposition in patients with essential hyper ension in vivo. Namely, activation of hepatic microsomal enzymes for oxidative metabolism was not affected by nifedipine.

Pressure ulcer is a kind of cutaneous and subcutaneous exelcosis and necrosis as suffered long-term pressure. Pressure ulcer mainly related with age, long-term in bed, chronic diseases, lower resistibility, vasculitis and less activities, as well as long-term pressing, poor blood circulation and untidy body environment. Pressure ulcer can be classified into 4 stages with severity. The risk to pressure ulcer should be evaluated, mainly with Braden score. Prevention of pressure ulcer included keeping clean, avoiding long-term pressure and find-ing potential pressure ulcer as soon as possible, based on improving the system situation and dealing with original diseases. For the patients suffered from pressure ulcer, it was important to decompress, debride the wound, keep humidity, and apply some new dressing with external drugs and vacuum sealing draining (VSD) and good psychological nurse and communication.

Objective To evaluate the specificity,sensitivity and clinical significances of direct antiglobulin test of bone marrow monouclear cells (BMMNC-Coombs test) and Flow Cytometry (FACS).Methods Autoantibodies on bone marrow mononuclear cells of 270 patients with immune and 471cases with primary or secondary anaemia and 19 normal controls were measured by BMMNC-Coombs test,and a part of IRP patients with negative BMMNC-Coombs test were retested by FACS.Results In 270 immune hemocytopenia patients,there were 163 (60.37%) with positive BMMNC -Coombs test.The positive rate of patients with immunorelated pancytopenia (IRP) was upmost and accounted for 67.2%.AIHA 63.6%,ITP 57.1%,Evans syndrome 43.2%.Both 471 patients with primary or secondary anaemia and 19 normal persons were all negative.In BMMNC-Coombs positive cases,there were mainly three types including IgG,IgG+C_3,IgG+IgM+IgA+C_3,they accounted for 30.4%,26.4%,20.8%.61 1RP with negative BMMNC-Coombs test were teased again for their BMMNC autoantibodies by FACS.83.6% of them were found to be positive with autoantibodies on BMMNC.The positive rate of autoantibodies on hematopoietic stem cells of IRP patients was 94.1%.it was higher than positive rate on any other types of bone marrow hematopoietic cells.Conclusion the sensitivity and specificity of BMMNC-Coombs test for diagnosis of autoimmune hemocytopenia was high,but the sensi- tivity of FACS assay was higher than that of BMMNC-Coombs test for detecting autoantibodies.Also had IRP,AIHA, ITP and Evans syndrome patients autoantibodies on bone marrow hematopoietic cells,The difference between IRP and other immune hemocytopenias was that IRP patients did have autoantibodies only on BMMNC.

The producing condition and properties of chitinase secreted by Nomuraea rileyi strain CQ031021 were studied.The optimal conditions for the strain to produce chitinase are 6 days of 28℃ with the initial pH 6.0,and the liquid medium containing 2.0% glucose as its carbon source and 0.6% peptone plus 0.6% beef extract as nitrogen source after inoculating dosage 2mL suspension of conidia(1?107/mL).The optimal temperature and pH for enzyme activity is 50℃ and 6.0,respectively,while the activity can be enhanced by Tween-80 and inhibited by SDS.The enzyme activity is stable under 40℃ and in pH range of 5.5～6.5.

Objective To evaluate the long?term effect of the comprehensive control strategy for schistosomiasis with empha?sis on infectious source control in marshland?type endemic areas of Poyang Lake. Methods Three heavy endemic villages with schistosomiasis in the lake areas were selected as pilots for the comprehensive control strategy which included replacing bovine with tractors and forbidding grazing on marshlands. The data of schistosome infections of residents and Oncomelania hupensis snails were collected and analyzed statistically from 2009 to 2014. Results There were no patients with schistosomiasis through feces examinations in the 3 pilot villages. The average density of live snails in Niuzhou marshland showed a decline trend and the density was stable in Liulingwei marshland. There were no infected snails in the 2 marshlands and no schistosome adult worms were found in the sentinel rats Conclusion The implantation of the comprehensive control strategy for schistosomiasis with em?phasis on infectious source control can control the prevalence of schistosomiasis during a long period.

Objective To comprehensively evaluate the clinical and radiological features of patients with Klebsiella pneumonia liver abscess (KLA) in order to improve treatment. Methods Data of 197 patients with KLA (n=106) or non-KLA (n=56) between March 2001 and January 2009 were collected and retrospectively analyzed on the aspects of underlying diseases, clinical manifestations, laboratory tests, pathogenic and imaging examinations. Results In comparison with non-KLA patients, the underlying diseases such as diabetes (53. 77% vs 25.00%, P = 0. 001) and hepatic adipose infiltration (16. 04% vs 5, 36%, P<0.05) were more common in KLA patients.Whereas the clinical presentations including abdominal pain (57. 14% vs 40. 57%, P<0. 05),hypodynamia (46.43% vs 19.81%, P=0. 001) and hepatomegaly (14.29% vs 4.72%, P<0. 05)were more severe in non-KLA patients than in KLA patients, however, the fasting blood glucose was higher in KLA patients than in non-KLA patients [(7.84±0.36) mmol/L vs (5.76±0.30) mmol/L,P=0. 001] on the admission. In addition, the abscess of KLA often appeared alone in the right lobe of the liver and was liable to generate air cavity (32. 88% vs 13.51%, P<0.05), un-smooth rim (71.23% vs 40.54% ,P<0.05) and dynamic septum enhancement (41.10% vs 16.22% ,P<0.01) in comparison with non-KLA. Conclusions Klebsiella pneumonia has emerged as the main pathogen of pyogenic liver abscess. The patients with KLA are often complicated with diabetes and fatty liver, as well as high prevalence of air cavity. The CT findings may be helpful for prompt treatment of Klebsiella pneumonia liver abscess.