AIM: To seek a small interfering RNA ( siRNA ) sequence targeting rat inhibitor of nuclear factor kappa Bα ( IκBα) that can specifically and effectively suppress IκBα mRNA expression of rat ciliary muscles in vivo.METHODS:Three IκBα specific double stranded siRNAs were designed and synthesized. They were transfected into rat A7r5 cells which express IκBα gene. Flow cytometry was used to assess transfected efficiency. The mRNA and protein levels of IκBα were examined by Real Time quantitative polymerase chain reaction ( Real Time-PCR ) and western blot to screen a candidate valid sequence with the highest inhibitory rate. The Cy3 labeled non-specific control siRNA or the candidate valid siRNA was then injected into rat anterior chamber. Distribution of Cy3- siRNA in rat ciliary muscles was viewed by fluorescence microscopy, and the inhibitory effect in vivo of the valid siRNA was identified via Real Time-PCR and immunofluorescence. RESULTS: The suppression effect of the siRNA targeting the CTACGATGACTGTGTGTTT of IκBα gene was most obvious by vitro screening. By anterior chamber injection, this valid siRNA could reach rat ciliary muscles and effectively suppress IκBα gene expression with the highest inhibitory rate of 59. 0% on mRNA level at 24h after RNAi, and 52. 3% on protein level at 72h after RNAi (P<0. 01).CONCLUSION: It proves that the siRNA targeting the CTACGATGACTGTGTGTTT of IκBα gene is the valid sequence to suppress rat IκBα expression of ciliary muscles by RNAi in vivo.

The type of basic media and the contents of plant growth substances were investigated by orthogonal design experiment,and also the effects of different culture conditions on the growth of suspension cells and the accumulation of total flavonoids in Eucommia ulmoides were studied.The results showed that B5 medium supplemented with 0.5mg/L NAA,0.6mg/L 6-BA and 30g/L sucrose,at initial pH 5.0～5.5,20g(FW)/L inoculation quantity and 110 r/min of rotation speed was a preferable culture conditions for E.ulmoides suspension cells growth and flavonoids synthesis.The results of metabolic kinetics analysis for E.ulmoides cell suspension culture showed that the logistic and Luedeking-Piret equations can be used for describing the kinetics of cell growth,sucrose consumption and flavonoids production during the process.The maximum specific growth rate(?m),the actual growth yield based on sucrose(YG) and maintenance coefficient(m) were 0.417/d,0.619g/g and 0.0206g/(g?d-1) respectively.All these outcomes could give a basis for establishing the suspension cell culture of E.ulmoides and production of the natural active components in large-scale.

Adult male Kunming mice were divided into normal control group,propylthiouracil(PTU) treated group and PTU+T_4 treated group.Neurogranin(Ng)protein expression in the hippocampus after 8 weeks was assayed by Western blotting and immunohistochemical method.Results showed that Ng protein was decreased in the hippocampus of male mice with adult-onset hypothyroidism,suggesting that decreased Ng,which is correctable by T_4,is involved in cognitive dysfunction in adult-onset hypothyroid male mice.

Objective To discuss the value and experience of the percutaneous vertebroplasty (PVP)in the treatment of vertebral body compression fracture(VCF)in aged osteoperosis.Methods PVP was performed in 44 cases with VCF including 28 with single vertebral compressed fracture,12 with double compressed fracture and four with triple compressed fracture,with 67 vertebrae,for clinical and radiologieal evaluation.Results The mean follow-up was 15 months(4-23 months).There could be seen immediate relief of pain in 40 cases,out-of-bed activities at operation day in 19 and out-of-bed activ- ities at second day after operation in 25.Postoperative X-ray showed uniformly distributed bone cement in the vertebral,without leakage.Conclusion PVP is a recommendable method for VCF,for it has ad- vantages of pain relief,vertebrae stabilization,minimal invasion and minor complications.

Objective To analyze the factors related to the occurrence of extensive cerebral infarction following surgeries for severe head injury and analyze the clinical outcomes of the patients. Methods Twenty patients with extensive cerebral infarction following surgeries for severe head injury were retrospectively analyzed for preoperative Glasgow Coma Scale (GCS) scores, hemorrhage volume, skull base fracture, pupil size, presence and duration of cerebral hernia. All the patients were treated with standard large bone flap decompression and/or routine comprehensive treatments. Results Patients with preoperative GCS score less than 5, intraeranial hemorrhage over 60 mL, and skull base factures complicated by prolonged cerebral hernia had significantly increased incidence of cerebral infarction. The 20 patients were followed up for 12 months and their clinical outcomes were evaluated with Glasgow Outcome Scale (GOS), which showed good recovery in 8 cases, moderate disability in 3 cases, severe disability in 2 cases, and vegetative survival in 3 cases. Death occurred in 4 cases. Conclusion Multiple factors may contribute to extensive cerebral infarction following surgery for severe head injury, among which low preoperative GCS score, massive intracranial hemorrhage, and prolonged skull base fracture with cerebral hernia are highly risk factors. Early detection of the infarction and timely management with decompression, dehydration, intracranial pressure control, promoting brain circulation, prevention of cerebral vasospasm, and mild hypothermia treatment may help lower the disability and mortality rates of the patients.

OBJECTIVETo study the clinicopathologic and immunohistochemical features of cystic neoplasms of the pancreas.

METHODSNinety-two cases of cystic neoplasm of pancreas were retrieved from the Department archival file during the period from 1999 to 2005. Histologic features were studied and the tumors were typed according to WHO classification. Immunohistochemistry was also carried out using paraffin-embedded tissues.

RESULTSThe age of patients ranged from 16 to 80 years. The patients included 33 males and 59 females. The tumors varied from 2 cm to 21 cm in diameter. They consisted of intraductal papillary mucinous neoplasm (36/92), serous cystic neoplasm (18/92), solid pseudopapillary tumor (18/92), mucinous cystic neoplasm (14/92), cystic pancreatic ductal adenocarcinoma (4/92) and cystic pancreatic endocrine neoplasm (2/92). Immunohistochemical study revealed variable staining patterns, with frequent overlaps between different tumor types. In general, serous cystic neoplasm expressed MUC1, while mucinous cystic neoplasm was positive for MUC-5AC, intraductal papillary mucinous neoplasm for MUC-2 and cystic pancreatic ductal adenocarcinoma for MUC-1. On the other hand, solid pseudopapillary tumor expressed alpha-antitrypsin, alpha-antichymotrypsin, vimentin and progesterone receptor.

CONCLUSIONSAccurate diagnosis of pancreatic cystic neoplasms requires correlation of clinical findings, radiologic examination, histologic features and immunostaining results. Pathologic distinction is important because of different prognostic significance. Two-thirds of pancreatic cystic neoplasms are premalignant or malignant and warrant surgical resection, whereas the remaining one-third (including pseudocyst and serous cystadenoma) are benign and can be treated conservatively.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Papillary ; metabolism ; pathology ; Cystadenocarcinoma, Mucinous ; metabolism ; pathology ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Cystadenoma, Mucinous ; metabolism ; pathology ; Cystadenoma, Serous ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Humans ; Male ; Middle Aged ; Mucin 5AC ; metabolism ; Mucin-1 ; metabolism ; Neoplasms, Cystic, Mucinous, and Serous ; metabolism ; pathology ; Pancreatic Neoplasms ; metabolism ; pathology ; Young Adult

OBJECTIVETo study the correlation between the expression of Egr-1 and NF-kappaB and the up-regulation of TNF-alpha and TGF-beta1 in macrophages after stimulation by silica in-vitro.

METHODSMacrophages were treated with antibodies against Egr-1 and NF-kappaB and antisense oligonucleotides. The level of TNF-alpha protein in the cell supernatant was then measured using enzyme-linked immunoadsorbent assay (ELISA). The expression of TGF-beta1 protein was detected by immunocytochemistry. The expression of TNF-alpha and TGF-beta1 mRNAs was also monitored by reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSCompared with silica-stimulated macrophages untreated with antibodies, the cells treated with 10 micro g/ml of Egr-1 or NF-kappaB antibodies were associated with reduced expression of TNF-alpha and TGF-beta1 proteins and mRNAs (P < 0.05). Compared with silica-stimulated untransfected group, the antisense group was associated with obvious reduction in the expression of TNF-alpha and TGF-beta1 proteins and mRNAs (P < 0.05).

CONCLUSIONThe expression of TNF-alpha and TGF-beta1 mRNAs and proteins are associated with activation of Egr-1 and NF-kappaB in macrophages, after stimulation by silica. It is possible that the corresponding antibodies and antisense oligonucleotides may become a potential therapeutic tool in the management of silicosis in the future.

Animals ; Antibodies ; immunology ; Cells, Cultured ; DNA-Binding Proteins ; genetics ; immunology ; Early Growth Response Protein 1 ; Immediate-Early Proteins ; genetics ; immunology ; Macrophages ; cytology ; metabolism ; Mice ; NF-kappa B ; genetics ; immunology ; Oligonucleotides, Antisense ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Silicon Dioxide ; pharmacology ; Silicosis ; etiology ; Transcription Factors ; genetics ; immunology ; Transforming Growth Factor beta ; biosynthesis ; genetics ; Transforming Growth Factor beta1 ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

AIM To investigate the distribution of volatile components and inorganic elements from Vernonia amygdalina Del..METHODS Volatile components and inorganic elements in different parts of V.amygdalina were analyzed and measured by HS-SPME-GC-MS and ICP-OES techniques.RESULTS Forty-four,sixty-seven,fifty-seven chemical compounds were identified from the root,stem and leaves of V.amygdalina,accounting for 83.9％,92.0％,83.9％ of the volatile components,respectively.Nineteen inorganic elements in total were detected,and the contents of As,Be,Bi,Co were too low to detect;The three inorganic elements with the highest content in root,stem and leaves were Mg,Al and Fe.CONCLUSION There are abundant volatile components and inorganic elements in V.amygdalina,with varying distribution in its different parts.

Objective To investigate effects of angelica polysaccharide on learning and memory abilities, Ach, ChAT, AChE, SOD, MDA in serum, APP and Aβ1-42 in hippocampus in model rats with Alzheimer disease (AD); To explore the mechanism of angelica polysaccharide for the treatment of AD. Methods Seventy SPF Wistar rats were selected for learning and memory ability by water maze. 10 rats were randomly selected (half female and half male) as sham-operation group, and the others were injected with Aβ25-35 by stereotatic techniques, copying AD model rats. 50 rats for learning and memory ability by water maze were successfully divided into model group, positive group, angelica polysaccharide low-, medium-, and high-dose groups, with 10 rats in each group. Rats in model group and sham-operation group were given normal saline for gavage, while rats in medication groups were given relevant medicine for gavage, 2 mL/(100 g?d), for 28 d. The learning and memory ability of rats in each group was tested by Morris water maze during 25-28 days, and the contents of Ach, ChAT, AChE, SOD, MDA in serum and APP and Aβ1-42 in hippocampus were determined. Results Compared with the sham-operation group, the escape latent period of model group was significantly prolonged in place navigation experiment; the target quadrant time was shortened; the latent time for the first time to reach the original escape platform was longer in spatial probe test; the residence time of crossing the original platform position and the target quadrant was shorter; the levels of Ach, the activity of ChAT and SOD in serum decreased; the levels of MDA, the activity of AChE in serum increased; the levels of APP and Aβ1-42 in hippocampus increased, with statistical significance (P<0.05, P<0.01). Compared with model group, the escape latent period of each medication group was shortened in different degrees after the intervention treatment; the residence time of target quadrant was prolonged; the latent time for the first time to reach the original escape platform was shortened; the number of cross platform increased; the levels of Ach, the activity of ChAT and SOD in serum increased; the levels of MDA and the activity of AChE in serum decreased; the levels of APP and Aβ1-42 in hippocampus significantly decreased, with statistical significance (P<0.05, P<0.01). Conclusion Angelica polysaccharide may effectively improve the learning and memory of ability of AD model rats to improve anti-free radical oxidation and promote Aβ metabolism and promote learning and memory ability of AD model rats, which have some preventive and therapeutic effects on AD.