Objective To observe the different effects of iodine excess on inducing two strain mice thyroiditis. Methods NOD and Balb/c mice, each having 14 mice, were divided into NaI and control group. The mice were given 0.05% NaI water for 8 weeks in NaI group. RIA and ELISA were used respectively to detect TT4, TgAb, TPOAb and TSH level in serum. Morphology changes of thyroid and apoptosis of thyrocytes stained by immunohistochemistry were observed under light microscope. Lymphocytic proliferation of cervical lymph node and spleen to responding to Tg were detected by MTr method. Results After intake of iodine water for 8 weeks, NOD and Balb/c mice showed relative quality of thyroid in Nal group[(104.83±14.52), (155.79±20.77)mg/kg]obviously increased compared with control group[(71.80±20.42), (105.15±21.98)mg/kg, t values:-3.293,-4.429, all P< 0.01)], enlarged follicular lumen with colloid accumulation were observed in thyroid. Serum level of TT4 in Nal group [(29.52±4.42), (19.53± 2.35)nmol/L]to control group[33.40±5.38), (23.47±6.22)nmol/L]of NOD and Balb/c mice showed a decreasing tendency(t values: 1.374,1.567, all P > 0.05). TSH of Nal group showed an increasing tendency in Balb/c mice[(4.14±1.71)μg/L, compared with control [(3.55±1.41)μg/L, t values:-0.705, P > 0.05]and obviously increased in NOD mice [(6.98±0.66)μg/L, compared with control[(555±056)μg/L, t values:-3.562, P< 0.01], but no change of TgAb and TPOAb level in Nal group(1281,1364 cpm, 2.50×103, 0.14×103U/L were observed, compared with control(1297,1220 cpm, 3.17×103,0.03×103 U/L; Zvalues:-0.081,-0.703, -0.244,-1.293, all P > 0.05). In NOD mice NaI group, apoptosis of thyrocytes was more intense than Balb/c mice, obvious infiltration of lymphoeytes, disorganization and focus fibrosis was seen in thyroid. The cell amount of NaI group increased in NOD mice lymph node and spleen cells[(1.100±0.014), (1.076±0.033)]were more than that in the control group [(0.993±0.011), (1.005±0.003), t value:-11.672,-4.314, P < 0.01). Conclusions Iodine leads to enlargement of thyroid and malfunction of thyroid in Balb/c mice. Besides, NOD mice have generate inflammatory reaction in thyroid and produced sensitized lymphocytes to Tg. Iodine excess can induce NOD mice to occur autoimmune thyroiditis.

The study is aimed to make the most of available space of Dendrobium officinale cultivation facility, reveal the yield and functional components variation of stereoscopic cultivated D. officinale, and improve quality, yield and efficiency. The agronomic traits and yield variation of stereoscopic cultivated D. officinale were studied by operating field experiment. The content of polysaccharide and extractum were determined by using phenol-sulfuric acid method and 2010 edition of "Chinese Pharmacopoeia" Appendix X A. The results showed that the land utilization of stereoscopic cultivated D. officinale increased 2.74 times, the stems, leaves and their total fresh or dry weight in unit area of stereoscopic cultivated D. officinale were all heavier than those of the ground cultivated ones. There was no significant difference in polysaccharide content between stereoscopic cultivation and ground cultivation. But the extractum content and total content of polysaccharide and extractum were significantly higher than those of the ground cultivated ones. In additional, the polysaccharide content and total content of polysaccharide and extractum from the top two levels of stereoscopic culture matrix were significantly higher than that of the ones from the other levels and ground cultivation. Steroscopic cultivation can effectively improves the utilization of space and yield, while the total content of polysaccharides and extractum were significantly higher than that of the ground cultivated ones. The significant difference in Dendrobium polysaccharides among the plants from different height of stereo- scopic culture matrix may be associated with light factor.
Cell Culture Techniques ; methods ; Dendrobium ; chemistry ; growth & development ; Plant Leaves ; chemistry ; growth & development ; Plant Stems ; chemistry ; growth & development ; Polysaccharides ; analysis

Objective To investigate the efficacy and clinical effectiveness of the steroid therapy with the all body and local application on hyperthyroid Graves ophthalmopathy.Methods A total of 57 patients (106 eyes)suffering from hyperthyroid Graves ophthalmopathy who hospitalized in the Department of Ophthalmology of the Second Affiliated Hospital of Harbin Medical University from Mar 2008 to Oct 2011 were enrolled in the study.Of the 57 patients,there were 21 males and 36 females,aged 18 to 57(average 37.28 ± 11.63).The clinical course was from 1 month to 5 years(average 29.63 ± 15.36 months).Based on the patient's age,sex and condition,the 57 patients were randomly divided into steroid pulse therapy with all body group(group SPT,29 patients,54 eyes)and peribulbar injection of long-term steraid group(group PI,28 patients,52 eyes).The patients in group SPT were treated with 0.5-l.0 g methylprednisolone by intravenous infusion daily for 3 days,and then stopped for 7 days,the treatment was repeated once.But the total application dose of methylprednisolone was not overruned 5.0-20.0 g.After that,the patients were treated with oral prednisone 40 mg/d.The dose of prednisone was reduced gradually to 5 mg/d.The total course of group SPT was 3 months.The patients in the group PI were treated with triamcinolone acetonide in periorbital injection of 20 mg once every two weeks for 3 months.After one month treated,all patients were visited.The patients' body weights of two groups were measured before the treatment and after one month treated.The ocular signs of the patients including visual acuity,diplopia,intraocular pressure,exophthalmos and horizontal visual palpebral fissure width were observed before and after the treatment.Results In the group SPT,the average body weight [(62.83 ± 7.23)kg]after the treatment was higher than that before the treatment[(57.03 ±7.13)kg],and also higher than that of the group PI[(59.89 ± 10.67)kg,t =25.25,2.08,all P＜ 0.05]after the treatment.In the group SPT,the average intraocular pressures before and after the treatment were (13.46 ± 1.78)and (13.69 ± 1.41)mm Hg(1 nm Hg =0.133 kPa),respectively.In the group PI,the average intraocular pressures before and after the treatment were (13.52 ± 2.19) and (13.77 ± 2.03)mm Hg,respectively.There was no significant differences before and after the treatment in both gr0ups(P＞ 0.05).In the group SPT and PI,the average exophthalmos after the treatment[(18.63 ± 2.22) and (14.85 ± 2.61)mm]were lower than those before the treatment[(20.78 ± 2.24) and (20.60 ± 2.81)mm,t =9.97,29.68,all P＜ 0.05],respectively.After the treatment,the average exophthalmos in the group PI was significantly lower than that of the group SPT(t =7.79,P＜ 0.05).In the groups SPT and PI,the average averagepalpebral fissure width after the treatment[(15.00 ± 1.80) and (11.65 ± 2.45)mm]were lower than those before the treatment[(17.22 ± 1.89) and (17.44 ± 2.37)mm,t =22.74,91.24,all P ＜ 0.05].After treatment of the group PI,the average exophthalmos was significantly lower than that of the group SPT(t =8.01,P ＜ 0.05).Conclusion Compared with the steroid pulse therapy with all body,the peribullbar application of long-term hormone for hyperthyroid Graves ophthalmopathy is more effective,with lcss side effects and worthy of promotion.

Nerve growth factor is one of the most important factors playing an important role in regulating the growth, development and survival of the neuron. The purified NGF from human placenta has been reported, the tissue from which can be isolated the NGF is very limited. It is important for basic research and clinic application to expression hNGF by genetic engineering. By polymerase chain reaction,gene fragment encoding the mature part of ?-NGF was amplified using the DNA of human placenta as template. The fragment was sequenced and inserted into expression vector pET-15b, and the recombinant expression vector pET15b-NGF was transformed into E.coli BL21(DE3)pLysS. After inducing with IPTG the NGF was higher expressed up to 25% of the total cell proteins. The expression product was purified with metal chelate affinity chromatography on Ni-IDA agarose under denaturing condition. The purity of rNGF was higher than 90% and yield of rNGF was 4.56mg/L expressing bacteria. SDS-PAGE revealed the NGF expression product had a Mr 16kDa. Western-blot displayed the recombinant product had strong immunological activity with rabbit anti-human ?-NGF polyclonic antibodies. The expression products were dealed with solubilizing inclusion bodies and refolding protein. The test of nerve fiber growth of chicken embryo DRG neurons displayed rNGF had biological activity.

Pseudorabies virus (PRV) is a swine herpesvirus of the Alphaherpesvirinae subfamily and a pathogen of swine resulting in devastating disease and economic losses worldwide. Cre/loxP site-specific system has the character of site specific, time specific, tissue specific and high efficiency in recombination, which makes this system universal in vivo and in vitro recombination of bacteria, fungus, plants, insects and mammals. A recombinant PRV which contain a loxP site in TK locus by using Cre/LoxP recombinant system was construsted. A pair of primers were synthesized according to the pEGFP-C1 sequence published on GenBank, and were used to amplify the EGFP gene expression cassette with two loxP sites flanking each side. This target gene was cloned into pSKLR, the resulting transfer vector pSKLR-GFP-loxP was then cotransfected into 293T cells with PRV SH strain genomic DNA. The recombinant virus rPRV1 was selected and purified in TK-143 cells by choosing fluorescent expressing plaques. Cre expression vector pOG231 was cotransfected into 293T cells with rPRV1 genomic DNA. The second recombinant virus rPRV2 was obtained, which contains only one loxP site in TK locus. Sequencing results of rPRV2 TK gene indicated that 34bp loxP site was inserted into rPRV2 genome and there were 270bp deletion in TK gene. PCR amplifying different generations of rPRV2 TK gene showed that the mutant was stable when passages in RK-13 cells. TCID_ 50 assay indicated that rPRV2 grows well on RK-13 cells. The LD_ 50 test results on BALB/C mice suggested that the virulence of rPRV2 was reduced. As a conclusion, the report gene GFP expression cassette was removed successfully from rPRV1 genome and only one LoxP site was leaved in rPRV2 genome by using Cre/LoxP recombinant system.

Objective To evaluate 99tcm-DTPA nuclear dynamic imaging in distinguishing the renal occupied disease.Methods A total of 164 in-patients with renal occupied disease who underwent surgery were included.According to the pathological diagnosis,119 patients had malignant tumors,and 45 patients had benign diseases.All patients’ imaging was retrospectively analyzed.Application of 99Tcm-DTPA nuclear dynamic imaging in renal occupied disease was compared with ultrasonography (US),computed tomography (CT),magnetic resonance imaging (MRI),intravenous pyelogram (IVP),and positron emission tomography (PET)-CT.Results The accuracy rates of different imaging methods in distinguishing between renal malignant and benign disease were 99Tcm-DTPA (84 ％,45 ％),US (72 ％,64 ％),CT ( 91％,92 ％),MRI (50 ％,67 ％),IVP (50 ％, 17 ％), respectively.The diagnostic accuracy rate of PET-CT for malignant tumors was 67 ％.The accuracy rates of 99Tcm-DTPA in distinguishing different phases of renal cell carcinoma were statistically significant (x 2 =83.4, P ＜ 0.01), while the accuracy rates in distinguishing renal cyst from renal angiomyolipoma were not statistically different.With the greater diameter, the diagnostic accordance rate is higher (x 2 =16.05,P ＜ 0.05).Conclusion 99Tcm-DTPA could be used not only to evaluate the renal function quantificationally,but also be helpful to distinguish renal malignant tumor from benign disease.

Objective: To observe the clinical efficacy of heat-sensitive moxibustion plus Western medicine in treating patients with diabetic peripheral neuropathy (DPN). Methods: A total of 70 patients with DPN were divided into an observation group and a control group by sealed envelope method combined with the random number table method, with 35 cases in each group. The control group was treated with routine medicine, and the observation group was treated with heat-sensitive moxibustion on the basis of the treatment in the control group. After 2 courses of treatment, the scores of Toronto clinical scoring system (TCSS) and vibration perception threshold (VPT) in both groups were observed, and the clinical efficacy was compared. Results: During treatment, 3 cases dropped out in the control group and 4 cases in the observation group. After treatment, the total effective rate in the observation group was higher than that in the control group (P<0.05). The scores of TCSS and VPT in both groups decreased after treatment, and the intra-group comparison showed statistical significance (both P<0.05). The scores of TCSS and VPT in the observation group were lower than those in the control group, and the differences were statistically significant (both P<0.05). Conclusion: Heat-sensitive moxibustion plus Western medicine can improve the symptoms in patients with DPN, and has a better curative effect than the Western medicine alone.

Objective To investigate safety managements of surgical instruments employed for the minimally invasive surgery through link management so as to guarantee surgery safety. Methods All procedure managements for surgical instruments were performed, such as distribution after sterilization, surgical use,takeover after the surgery for 1 910 cases from July to December 2009 in our hospital. All the surgical instruments involved in the surgery included thoracoscopes, laparoscope, uteroscope, ventriculoscope and surgical robots. We organized management training of surgical instruments for medical staff, filed out management procedures,enhanced their consciousness of safety management, controlled the key link for quality control, performed gap management, handled the existing problems and improved management quality. Results The safety procedure management of surgical instruments was performed for the 1 910 cases, which insured that the surgery could go smoothly. Conclusions Safety management measures are established on the basis of the information traced from surgical use, takeover after the surgery, cleaning, sorting, inspection for surgical instruments so that oversight over its management could be effectively avoided, making its clinical effects more obvious.

Objective To investigate the clinical and mammographic features of plasma cell mastitis.Methods Twenty-five patients(28 lesions)with histologically confirmed plasma cell mastitis, aged from 26 to 70 years(mean age 41 years),were examined with X-ray mammography.The clinical manifestations and imaging features were retrospectively reviewed.Results No case was in lactation.The painful irregular masses,ranged from 1.3 to 8cm in size,were found in 22 patients,while 3 patients with acute episode.Recurrent episodes of breast masses were noted in 4 patients.Based on the mammographic appearances,the plasma cell mastitis were classified as the following four types:inflammation-like type (2/28),ductal ectasia type(3/28),focal infiltration type(10/28)and nodular type(13/28).The valuable radiogyaphic signs:(1)An asymmetrically increased density along the lactiferous duct with a flame-like appearance,inhomogeneous low density tubular structures and scattered stick-shape calcifications.(2) Architectural distortion and oil cysts formation in adjacent area,(3)Subareolar ductal ectasia.Conclusions The clinical and mammographic characteristics of plasma cell mastitis are critical to avoiding unnecessary surgery.Histopathological result is needed for the diagnosis in patients highly suspected of malignancy.

OBJECTIVESTo investigate whether antisense Bmi-1 plasmid could inhibit the proliferation of Jurkat cells.

METHODSThe antisense plasmid was constructed by PCR amplification of a 171 bp segment spanning Bmi-1 start codon and zinc finger structure and the PCR product was subsequently inserted reversely to plasmid pLNCX2. The final construct was confirmed through restriction enzyme digestion. G418 was added into the medium after the plasmid was successfully introduced into Jurkat cells by using lipofectin-mediated DNA transfection. The proliferation of Jurkat cells were determined by MTT and colony formation assays. Cell cycle was determined by flow cytometry. The p16 expression of Jurkat cells was studied by immunofluorescent histochemistry.

RESULTSThe growth rate of antisense Bmi-1 transfected Jurkat cells was significantly lower than that of the controls, and the colony forming capacity of the transfected cells decreased significantly (P < 0.01), the colony numbers being (90.7 +/- 9.07)/10(3) cells, (83.3 +/- 6.11)/10(3) cells and (56.0 +/- 5.56)/10(3) cells for control cells, empty plasmid transfected Jurkat cells and antisense Bmi-1 transfected Jurkat cells, respectively. The percentage of G, phase cells was increased and the p16 expression of antisense Bmi-1 transfected cells was significantly upregulated than that of control cells.

CONCLUSIONAntisense Bmi-1 can inhibit the growth and upregulate the expression of p16 of Jurkat cells in vitro.

Cell Cycle ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; Jurkat Cells ; Nuclear Proteins ; genetics ; Oligonucleotides, Antisense ; genetics ; Plasmids ; genetics ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; genetics ; Repressor Proteins ; genetics ; Transfection