AlM:To construct recombination eukaryotic expression plasmid of human thyrotropin receptor extracellular domain encapsulated with cationic liposomes.
METHODS:We amplified the target gene of shuttle vector PHMCMVTSHR289, conjugated the target gene and eukaryotic expression plasmid pcDNA3. 1 +, and accredited whether pcDNA3. 1+/TSHR289 was connected or not by enzymatic digestion and sequencing. Cationic liposomes encapsulated the recombination plasmid pcDNA3. 1+/TSHR289.
RESULTS: Recombination plasmid pcDNA3. 1+/TSHR289 digested with enzyme Hindlll and the fragment through 0. 8% gel electrophoresis showed 512bp strip. Recombination plasmid pcDNA3. 1+/TSHR289 were found synonymous mutation through forward ( AAC to AAT ) and reverse sequencing ( GCG to GCT) . The volume ratio of cationic liposomes and recombinant plasmid was 3:1.
CONCLUSlON: lt is successful to construct the recombination plasmid pcDNA3. 1+/TSHR289 by accredit it through enzymatic digestion and sequencing.

Background As one of the most common microvascular complication of diabetes in eyes,diabetic retinopathy (DR) is one of the most important cause of blindness.Nuclear factor-kappa B (NF-κB) is involved in the occurrence and development of the disease through the activation of a series of inflammatory cytokines.Objective The present study was to investigate the effects of peroxisome proliferator-activated receptor-gamma (PPAR-γ) excitomotor,rosiglitazone,on NF-κB expression and apoptosis of the retinal ganglion cells (RGCs) in the retina with diabetes mellitus. Methods Ninety SPF male Wistar rats were randomized into normal control group,diabetic control group and rosiglitazone group.Diabetes mellitus was induced by intraperitoneal injection of 50 mg/kg streptozotocin(STZ).Then 3 mg/kg rosiglitazone was intragastricly administered once per day in the rosiglitazonegroup,and the same volume of saline solution was used at the same way in the normal control group and diabetic control group from 3 days after modeling.The rats were sacrificed and the eye cups specimen was made at 4,8 and 12 weeks after usage of drugs.Retinal histopathological examination was performed by hematine-eosin staining,and expression of NF-κB p65 protein in retina and apoptotic index(AI) of RGCs were detected by immunohistochemistry and TUNEL assay,respectively in different time points mentioned above.The use of the animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State and Technology Commission.Results The blood glucose level was significantly elevated at various time points in the diabetic control group and rosiglitazone group compared with normal control group (P＜0.01 ),and that of the rosiglitazone group was significantly declined in comparison to the diabetic control group (q =0.81,0.82,1.23,P＞ 0.05 ).Normal retinal structure was seen in the normal control group,and edema retinal cell and disorder of retinal layers were exhibited in the diabetic control group.Retinal structure was almost normal in the rosiglitazone group.The NF-κB p65 was expressed weakly in the retina of normal control group,but the expression of NF-κB p65 was significantly elevated in the diabetic control group and rosiglitazone group compared with the normal control group(P＜0.01 ).However,the expression of NF-κB p65(A value)was significantly decreased in the rosiglitazone group compared with diabetic control group at 8 weeks and 12 weeks( q=17.77,15.30,P＜0.01 ).There were a few apoptotic cells in rat retina of the normal control group.Compared with the normal control group,the AI of the diabetic control group and rosiglitazone group was significantly reduced(P＜0.01 ).However,the AI of RGCs in the rosiglitazone group was significantly lower than that of diabetic control group in various time points (q =19.28,27.39,49.92,P＜0.01 ). Conclusions As one of the PPAR-γexcitomotors,rosiglitazone can inhibit apoptosis of RGCs through downregulating the expression of NF-κB in rat retina with diabetes mellitus,indicating a protective effect of rosiglitazone on retina in diabetic rat.

AlM:To observe the changes of binocular vision in V-pattern exotropia children before and after surgical correction, and the effect of training in reconstructing the binocular vision after surgical corrections.METHODS: Sixty V-pattern exotropia children were enrolled in this study and were divided into three groups according to their age:group A (4~6 years old), group B (7~9 years old), and group C (10~12 years old), 20 cases for each group. Patients received routine refraction and ophthalmic examinations. Distance and near deviation were measured by prism-covering method and synoptophore. The simultaneous perception and fusion were examined with a synoptophore, and the stereacuity was measured with stereograms ( Titmus) . The children who didn’t reconstruct binocular vision function 1wk after surgery received binocular vision training. The data were recorded before and 1 , 2, 4, and 8wk after surgery. RESULTS: Binocular vision significantly improved among the children after surgery in group A and B ( P<0. 05 ) . Significantly divergence showed between group C and the other groups 1wk after surgery ( P < 0. 05 ). Binocular vision of the three groups all significantly improved 8wk after surgery, with no significant differences (P>0. 05). CONCLUSlON: V - pattern exotropia children can benefit from early surgical correction and training after surgery in reconstruct binocular vision.

Objective To investigate the expression of 6-phosphofructo-2-kinase /fructose-2,6-bisphosphatase-3 (PFKFB3) and sphingosine 1-phosphate receptor 2 (S1 P2) in the retina of rats with type 2 diabetes mellitus (DM) and to explore the correlation of tertiary butylhydroquinone (tBHQ) with PFKFB3 and S1P2.Methods Together 60 male SD rats were divided into normal control group (NC group),diabetes mellitus group (DM group) and tBHQ group.Type 2 DM model was induced in the latter two groups.The rats in tBHQ group were given 10 g · L-1 tBHQ in high-fat and highsugar diet 1 week after successful modeling,while DM rats were fed with high-fat and high-sugar diet continuously.At 4 weeks and 12 weeks after tBHQ intervention,blood samples were taken from the hearts of rats in each group,and serum contents of fasting plasma glucose (FPG) and fasting serum insulin (FINs) were measured.Immunohistochemistry and qRT-PCR were taken to detect the distribution and expression of PFKFB3,S1 P2 and vascular endothehal growth factor (VEGF) mRNA and protein in the retina,and TUNEL methods were used to detect apoptosis index of retinal ganglion cells of rats in each group.Results There were significant difference in the FPG and FINs levels of the three groups at 4 weeks and 12 weeks (all P =0.000).Light microscopy test showed that positive expressions of PFKFB3,S1 P2 and VEGF protein were found in all groups at 4 and 12 weeks,which were mainly located in the retinal ganglion cell layer and the inner nuclear layer.Immunohistochemistry and qRT-PCR showed that the difference in relative expressions of PFKFB3,S1P2,VEGF protein and mRNA in the retina at different times after modeling were statistically significant (all P ＜ 0.05).At 4 and 12 weeks,the expression levels of PFKFB3,S1P2 and VEGF in DM group were higher than those in NC group,but their expressions in tBHQ group were significantly downregulated when compared with DM group,and all differences were statistically significant (all P ＜ 0.05).Compared with 4 weeks,PFKFB3,S1 P2 and VEGF mRNA and protein in DM group were overexpressed at 12 weeks,and the expression level of S1 P2 in tBHQ group at 12 weeks was lower than that at 4 weeks,and the differences were statistically significant (all P ＜ 0.05).TUNEL assays showed that there was significant difference in the apoptotic index (AI) of retinal ganglion cells of rats in each group at different times after modeling (all P ＜ 0.05).DM group had higher AI than NC group (P ＜ 0.05),and tBHQ group was lower than DM group (P ＜ 0.05);Compared with 4 weeks,DM group had increased AI at 12 weeks while tBHQ group had decreased AI (both P ＜ 0.05).Conclusion PFKFB3,S1P2 and VEGF are involved in the pathological process of the retina of type 2 DM rats,which may play a role through the PFKFB3 / VEGF / S1P2 signaling pathway and may be related to the apoptosis of retinal ganglion cells.And it is indicated that tBHQ has a protective effect on the retina of type 2 DM rats.

Pharmaceutical preparations, particularly as a "secret recipe" of traditional Chinese medicine in medical institutions, are the product of China's medical and health industry, and they are also an important means of competing of different medical institutions. Although pharmaceutical preparations have advantages and characteristics than institutes for drug and pharmaceutical companies, the quality standards of pharmaceutical preparations in medical institutions has not reached the desired level over the years. As we all know, the quality of pharmaceutical preparations is important to ensure the efficacy, especially under the environment of people pay more sttention on drug safety and effectiveness and contry increase emphasis on the stste of pharmaceutical preparations. In view of this, we will improve the grade, stability, and clinical efficacy of pharmaceutical preparations by the advanced equipment, testing instruments and the process dynamic quality control technology. Finally, we hope we can provide new ideas for the quality control of pharmaceutical preparations.
Drug Compounding ; standards ; Medicine, Chinese Traditional ; standards ; Quality Control

AIM: To establish an experimental model of high intraocular pressure in mice by laser photocoagulation and to prepare for future research.
METHODS: Experimental model of high intraocular pressure was induced unilaterally in 44 C57BL/6 mice. The fellow eye served as a control. TONO-PEN AVIA Tonomter was used to measure intraocular pressure (IOP) to guarantee IOP value at 1, 2, 4, 8wk. Slit-lamp biomicroscopy was performed throughout the period and the structural changes were assessed histologically. And then, their eyes were enucleated, postfixed, cryoprotected, and embedded in optimal cutting temperature medium. After hematoxylin and eosin stain ( HE stain ) , cryosections of the retina were observed under light microscope. TdT-mediated biotin-dUTP nick end labeling ( TUNEL ) was performed on the retinal sections to determine apoptosis rate.
RESULTS: IOP of laser-treated eyes was significantly higher than that of control eyes from 1-8wk (P<0. 05). The highest IOP was 31mmHg, but only one eye. The IOP was mainly around 20mmHg. In laser-treated eyes, the angle of anterior chamber were narrow. Number of cells in the inner nuclear layer and retial gangllion cell layer was slightly lower than that in control eyes at 2wk, but by 4 and 8wk the number of cells was significantly lower than that in the control contralateral eyes.
CONCLUSION: The laser photocoagulation of limbus causes chronic elevation of IOP and this method may be a promising experimental model for the investigation of biological mechanisms of glaucomatous retinal ganglion cell damage.

Objective To observe the effect of human immunoglobulin on elderly head and facial post herpetic neuralgia (PHN) and peripheral blood TNF-α.Methods One hundred and twenty-two inpatients with PHN aged ≥65 years old were selected and divided into the observation group (52 cases) and control group (70 cases) by the systematic sampling method.The control group was given the early conventional combined therapy,while on this basis the observation group was intravenously dripped by human immunoglobulin.The incidence rate of PHN and pain visual analog scale(VAS) score at 1,2,3 months after recovery discharge from hospital were recorded in the two groups.Other 20 healthy elderly people were selected as the healthy control group.The TNF-α level was determined in the two patients groups before and after treatment and in the healthy control group.Results The PHN occurrence rate at 1,2,3 months after discharge in the observation group was significantly lower than that in the control group(P＜0.05);the VAS score after treatment and at 1,2,3 months after discharge in the observation group was significantly lower than that in the control group (P＜0.05);serum TNF-α level after treatment in the observation group was significantly lower than that in the control group (P＜0.01),moreover which was close to the level in the healthy control group(P＞0.5).Conclusion Human immunoglobulin can reduce the PHN occurrence in the old people with head and facial herpes zoster and reduce the peripheral blood TNF-α level.

OBJECTIVETo investigate the association between perineural invasion(PNI) and clinicopathological factors and the effect of PNI on overall survival in patients with gastric carcinoma.

METHODSA total of 178 patients with gastric carcinoma from January 2004 to May 2008 were analyzed retrospectively. Paraffin sections of surgical specimens from all the patients who underwent gastric resection were stained with laminin. PNI-positive was defined as infiltration of carcinoma cells into the perineurium or neural fascicles. The association of PNI with clinicopathologic features and prognosis of gastric carcinoma was studied.

RESULTSPNI was positive in 78 of 178 patients(43.8%). The proportions of T stage, lymph node metastasis and TNM stage were significantly higher in PNI-positive group than those in PNI-negative group(all P<0.01). The PNI positive rate was correlated with the depth of gastric mural invasion and clinical stage. The overall survival in PNI-positive group was significantly lower than that in PNI-negative group by univariate analysis(P<0.01). The mean survival of PNI-positive patients(28.6 months) was significantly shorter than that of PNI-negative patients (44.3 months, P<0.01), which was also influenced by pN stage, pT stage, and clinical stage(P<0.01). By multivariable Cox proportional hazards model of overall survival, the positivity of PNI appeared to be an independent prognostic factor (hazards ratio=2.257, 95% CI:1.268-4.019, P=0.006).

CONCLUSIONSPNI is associated with the degree of malignancy in gastric cancer. PNI can be a candidate of new prognostic factor.

Aged ; Carcinoma ; pathology ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Peripheral Nerves ; pathology ; Prognosis ; Retrospective Studies ; Stomach ; pathology ; Stomach Neoplasms ; pathology

OBJECTIVETo investigate the relationship between the extracellular matrix (ECM) and neoplastic progression in hamster with tongue cancer.

METHODSForty-eight specimens of hamster tongue cancer were divided into control group (n=6) and experimental group (n=42). The pathological grade of the specimens was assessed (including 3 stages, namely atypical hyperplasia, carcinoma in situ and early invasive carcinoma). The sections of the tongue were stained with Masson and aldehyde-fuchsin (AF) staining for microscopic observation of the elastic fiber and collagen fiber changes.

RESULTSWithin the connective tissue cores (CTC) of the papillae in the control group was a framework of numerous and fine Gomrori's aldehyde fuchsin-positive elastic fibers. But in the stages of dysplasia and carcinoma in situ, these elastic fibers decreased and further diminished in the CTC in early invasive carcinoma. In dysplasia and carcinoma in situ stages, most of the elastic fibers collapsed with scattered elastic fibers, and the elastic fibers decreased significantly in early invasive carcinoma. The control group showed a significantly greater number of elastic fibers in the experimental group. The collagen fiber was obviously increased and irregularly arranged in dysplasia and carcinoma in situ stage; in early invasive carcinoma, the collagen fibers became thicker with deposition in the lamina propria.

CONCLUSIONAn excessive deposition of collagen fiber and reduction of the elastic fibers is an important factor contributing to the development of tongue carcinoma in hamsters.

Animals ; Carcinoma ; pathology ; Collagen ; metabolism ; Connective Tissue ; pathology ; Cricetinae ; Elastic Tissue ; pathology ; Extracellular Matrix ; pathology ; Neoplasms, Experimental ; pathology ; Tongue Neoplasms ; pathology

OBJECTIVETo study the efficacy and safety of Naoxintong capsule treatment of stroke recovery with Qi-deficiency and blood-stasis syndrome (cerebral infarction), and to compared the non-inferiority analysis with the positive drug Tongxinluo capsule.

METHODTaking Tongxinluo capsules as control, randomized, double-blind, controlled, multi-center clinical experiments were studied. The evaluating indexes included the decrease of integral value of stroke patients, changes in traditional Chinese medicine, the improvement of the patient viability status (disability level), Chinese stroke scale (CSS), activities of daily living (DAL) scale and barthel index (BI ) points.

RESULTThe total effect of the two groups, Chinese and other symptoms, showed no significant statistical significance.

CONCLUSIONNaoxintong capsule stroke recovery, with Qi-deficiency and blood-stasis syndrome (cerebral infarction) has a therapeutic effect, and more secure.

Activities of Daily Living ; Adult ; Aged ; Capsules ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Qi ; Stroke ; drug therapy ; Treatment Outcome