Objective　To study the indication, contraindication and short-term clinical effects of stereotactic radiotherapy (X-knife) in lung cancer. Methods　A total of 56 cases (Treatment group) of lung cancer were treated with X-knife composing of the modern 3-D treatment planning system and the stereotactic body frame, and SL-75 accelerator. And another 53 cases only treated with routine external radiation served as the control. In treatment group, 42 cases received 50 Gy radiation, at 2 Gy per day, 5 d per week in the upper mediastinum, hilus of lung, below the primary focus first and then underwent X-knife for the primary focuses, and other 14 recurrent and metastatic cases received irradiation of 8-12 Gy per day, 5 d by X-knife alone. Results　The actually reexamination rate for 1, 3, 6, and 12 months after X-knife treatment in 78 focuses from 56 cases was 79.5% (62/78), 89.8% (70/78), 87.2% (68/78) and 78.2% (61/78) respectively. The focuses reappeared in 3 months after the treatment and the growth rates of focus in 3, 6, and 12 month after the treatment were 2.6% (2/78), 5.1% and 9% (7/78) respectively. There were 2 focuses received a second treatment with X-knife because no change after the first one. Both the 1-and 2-year survival rates of 36 cases of the primary lung cancer in the treatment group were higher than that of control group (89.1%, 53.9% and 77.4%, 39.6%). Conclusion　The stereotactic radiotherapy is of applicable and prospective in the treatment of lung cancer. The indications include: ①As a boost dose in case when primary focus is less than 5 cm after the first external radiotherapy may reduce the exposure of lung tissue to x-ray and the occurrence of radiation pneumonia; ②Treatment for the recurrence after radiotherapy and operation; ③Radical treatment for the metastatic focus less than 5 cm. Attention must be paid to when multiple treatment is carried on the volume of the focus, the Karnofsky scores and the general condition when stereotactic radiotherapy combined with X-knife for lung cancer.

To establish a MacELISA method for the detection of IgM antibodies against Chikungunya virus (CHIKV), we prepared virus like particle (VLP) antigens of CHIKV using the whole structural protein C-E3-E2-6K-E1 encoding gene with a baculovirus expression system in Sf9 insect cells. The VLPs were purified and used to immunize Kunming mice. Then, polyclonal antibodies were purified from the samples of ascites with a protein G HiTrap SP column and labeled with horseradish peroxidase. A MacELISA method for the detection of IgM antibodies against CHIKV was assembled with goat anti-human IgM antibody, VLP antigens and an enzyme-labeled polyclonal antibody. The results were evaluated with a serum panel containing serum samples from laboratory-confirmed CHIK, HFRS patients, healthy donors, and commercially available CHIKV IgM as a quality control. It was shown that the MacELISA had a specificity of 99% (99/100), the coefficients of variation (CoV) within a plate were <10%, and the CoV of different ELISA plates in terms of the plate variation coefficient was <15%. A comparative analysis was performed to compare the current method against a commercial CHIKV IgM antibody detection kit for IIFA-IgM. The detection limit of MacELISA was significantly lower than that of the IIFA-IgM commercial kit (P< 0.0001). Here, we demonstrate that the VLP-based MacELISA is a promising tool for the early diagnosis and epidemiological investigation of CHIKV infection, with a high level of sensitivity and specificity for the detection of IgM antibodies against CHIKV.
Animals ; Antibodies, Viral ; blood ; Chikungunya Fever ; blood ; diagnosis ; virology ; Chikungunya virus ; immunology ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunoglobulin M ; blood ; Mice

The effects of different physical and chemical factors on hairy root growth and flavonoids production were studied in suspension culture of Saussurea medusa hairy root in 1/2 MS medium. The results showed that the following culture conditions, nitrogen concentratiaon (involved NH4+ and NO3-), 30 mmol/L; the ratio of ammonium to nitrate, 5:25; the combination of 2% sucrose and 3% glucose; 0.5 mg/L GA3; 0.5 mg/L IBA; initial pH 5.8; light cycle, 18 h/d (3500lx); temperature, 24 degrees C; shaker revolutions per minute, 100 r/min, were favourable to hairy root growth and flavonoids production. Under the above culture conditions, up to 12.8 g/L (DW) of hairy root and 1922 mg/L of flavonoids were obtained after 21 days of culture. The content of total flavonoids in hairy root was 15%, which was about 25 times as that in the wild plantlet.
Culture Media ; Flavonoids ; biosynthesis ; Plant Roots ; growth & development ; metabolism ; Saussurea ; growth & development ; metabolism ; Temperature ; Tissue Culture Techniques

OBJECTIVETo evaluate the feasibility and utility of an ex vivo sentinel lymph node (SLN) identification and ultrastaging for colorectal cancer (CRC).

METHODSCRC patients undergoing resection of a primary colorectal cancer were considered for inclusion. Following resection, SLN identification was performed. The SLN was dissected from the mesentery and submitted separately for pathologic analysis. All lymph nodes were stained with HE. Blue lymph nodes, when negative by routine HE staining, were further analyzed.

RESULTSA total of 62 tumors from 60 patients with colorectal cancer were studied. 95.2% (59/62) specimens was successfully identified. In these 59 specimens, a total of 1114 (18.9 per specimens) lymph nodes were examined; of these, 157 (14.9%) were designated as SLNs. The number of blue-stained lymph nodes removed ranged from 1 to 9, with a mean of 2.7 blue nodes identified. The sensitivity of a blue-stained lymph node identifying metastatic disease was 39.1%. The false-negative was 23.7%. In 4 specimens micrometastases were detected only by immunohistochemistry with cytokeratin.

CONCLUSIONSEx vivo sentinel lymph nodes mapping in colorectal cancer is feasible and can identify the SLNs with a very high success rate. Ex vivo SLN mapping improves pathologic staging of patients with CRC. The SLN evaluation should not replace attempts to harvest large number of nodes for standard processing. SLN mapping can help improving the number of nodes for pathological examination.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; pathology ; Female ; Humans ; Immunohistochemistry ; Keratins ; analysis ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Rosaniline Dyes ; Sentinel Lymph Node Biopsy ; methods

Animals ; Bone Marrow Cells ; cytology ; Brain ; cytology ; physiopathology ; Brain Injuries ; physiopathology ; therapy ; Cell Movement ; Cells, Cultured ; Cisterna Magna ; Immunohistochemistry ; Rats ; Rats, Sprague-Dawley ; Stromal Cells ; transplantation

Objective To detect the mutations of ATP2C1 gene in patients with Hailey-Hailey dis- ease (HHD).Methods PCR and direct sequencing were performed in 17 patients and 120 healthy controls to screen the mutations in the exons of ATP2C1 gene.Results Eight mutations were identified in nine probands, including three deletion mutations (nt1464-1487 del/nt1462-1485del,1523delAT,2375delTTGT),three splice site mutations (360—2A→G,1415—2A→T,2243+2T→C) and two missence mutations (C920T and G1942T).None of the above mutations was found in the controls.Conclusion Eight specific novel mutations were identified in nine probands of HHD,which could be causative factors of the disease.

OBJECTIVETo study the immunohistochemical expression of OCT4, CD117 and CD30 in germ cell tumors and to assess their diagnostic value.

METHODSImmunohistochemical study for OCT4 was performed on formalin-fixed, paraffin-embedded tissues of 63 cases of germ cell tumors, including seminoma (21), dysgerminoma (7), germinoma (8), embryonal carcinoma (8), yolk sac tumor (6), mature teratoma (10) and immature teratoma (3), as well as 25 cases of non-germ cell tumors, including granulosa cell tumor (8), clear cell adenocarcinoma (4), Leydig's cell tumor (5), diffuse large B-cell lymphoma (4) and malignant melanoma (4). Besides, the expression of CD117 and CD30 in all germ cell tumors was studied.

RESULTSAll cases of seminoma and germinoma, 6/7 cases of dysgerminoma and 7/8 cases of embryonal carcinoma were positive for OCT4, with strong nuclear staining. All other germ cell tumors and non-germ cell tumors were negative for OCT4, except for 1 case of yolk sac tumor and 1 case of clear cell adenocarcinoma which showed weak staining. Positive membranous expression of CD117 was demonstrated in 19/21(90.5%) seminoma, 5/7 dysgerminoma and 7/8 germinoma. Focal weak membranous staining was also noted in 1 case of yolk sac tumor. The melanocytes in teratoma were also positive for CD117. All cases of embryonal carcinoma were negative. On the other hand, positive membranous expression of CD30 were demonstrated in 6/8 embryonal carcinoma. One case of germinoma and 1 case of yolk sac tumor showed weak cytoplasmic positivity. All cases of seminoma and dysgerminoma, 7/8 germinoma and all cases of teratoma were negative for CD30.

CONCLUSIONSOCT4 is a sensitive and relatively specific marker for diagnosing seminoma, dysgerminoma, germinoma and embryonal carcinoma. CD117 and CD30 immunostains, when used in combination, represent valuable tools for distinguishing embryonal carcinoma and seminoma, dysgerminoma, germinoma.

Carcinoma, Embryonal ; metabolism ; pathology ; Diagnosis, Differential ; Dysgerminoma ; metabolism ; pathology ; Endodermal Sinus Tumor ; metabolism ; pathology ; Female ; Germinoma ; metabolism ; pathology ; Humans ; Ki-1 Antigen ; metabolism ; Male ; Neoplasms, Germ Cell and Embryonal ; metabolism ; pathology ; Octamer Transcription Factor-3 ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-kit ; metabolism ; Seminoma ; metabolism ; pathology ; Teratoma ; metabolism ; pathology ; Testicular Neoplasms ; metabolism ; pathology

A full-length cDNA encoding a MYB-related regulatory gene was isolated from a cDNA library prepared from mRNAs of the red line callus of S. medusa by TD-PCR. The cDNA, designated SmP, is 969 nucleotides long and has an open reading frame of 771 bp with a deduced amino acid sequence of 256 residues. The putative protein of SmP has two typical conversed R2R3-Myb DNA-binding domains in N-terminal and displays a rather high degree of similarity to OsMYB from rice and LBMI from tobacco, showing 73% and 70% identity within the DNA-binding domains. However, the C-terminal domain of the SmP protein does not show obvious similarity to any other known protein sequence. It is rich in hydrophilic amino acids, especially in serine residues (18.38%), partly organized in homopolymeric stretches, a feature often found in activation domain of transcription factors.
Amino Acid Sequence ; Cloning, Molecular ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; classification ; genetics ; metabolism ; Polymerase Chain Reaction ; Saussurea ; classification ; genetics ; metabolism ; Sequence Homology, Amino Acid ; Transcription Factors ; chemistry ; genetics ; metabolism

OBJECTIVETo investigate whether mammalian target of rapamycin (mTOR) kinase was abnormally activated in maldeveloped balloon cells and dysmorphic neurons of focal cortical dysplasia (FCD) with refractory epilepsy.

METHODSA total of 12 archival cases of FCD typeIIwith medically intractable epilepsy treated between 2008 and 2010 were retrieved. Perilesional brain tissue was used as control specimens (n = 8). The expression of phosphorylated p-AKT (Ser473), p-mTOR (Ser2448) and p-P70S6K (Thr389) was investigated by imunocytochemistry.

RESULTSThe expression of p-AKT (Ser473), p-mTOR (Ser2448) and p-P70S6K (Thr389) was found in meldeveloped balloon cells and dysmorphic neurons of FCD. A weak stain in a small amount of pyramid neurons was also found in the control group.

CONCLUSIONAbnormal activation of mTOR in maldeveloped balloon cells and dysmorphic neurons of FCD may be a key molecular mechanism underlying the histological changes and repeated seizures.

Adolescent ; Adult ; Brain Diseases ; metabolism ; pathology ; Child, Preschool ; Epilepsy ; metabolism ; pathology ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Immunohistochemistry ; Male ; Malformations of Cortical Development ; metabolism ; pathology ; Malformations of Cortical Development, Group I ; Nestin ; metabolism ; Neurons ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Ribosomal Protein S6 Kinases, 70-kDa ; metabolism ; TOR Serine-Threonine Kinases ; metabolism ; Young Adult

Hairy root clones of Saussurea involucrata transformed with Agrobacterium rhizogenes strains R1601, R1000, and LBA9402 were established to investigate the flavonoid production. Opine synthesis and PCR analysis confirmed the integration of the T-DNA fragment of Ri plasmid from A. rhizogenes strain R1601 into the transformed root genome. The frequency of hairy root formation from root segments, which were pre-cultured 2 days in N6 solid medium without plant growth regulators, amounted to 100% following infection with R1601 strain of A. rhizogenes. The transformed roots were kept in hormone-free N6 liquid medium in the dark at 25 degrees C, 110r/min and routinely subcultured every 20 - 24 days. One hairy root clone, which grew vigorously with lateral branches, was periodically examined for the ability to produce flavonoid. The maximum of biomass and flavonoid yield achieved 66.7 g/L (fresh weight) and 102.3mg/g dry weight after incubation 20 days. The calli were induced from the hairy root culture in the presence of 0.5mg/L IBA and intact plantlets were regenerated from these calli. The regeneration plantlets from hairy roots, in which the flavonoid content were 53% in that of untransformed plants, weren't different in growth and morphology of the untransformed plantlets. Therefore plant regeneration from hairy roots may be also a means for producing transformed S. involucrata plants. Hairy root cultures of S. involucrata clearly showed higher flavonoid contents compared to the wild plant or the regeneration seedlings. As the wild S. involucrata grows only in special regions with peculiar climate, and cultivation of this species in a normal climate has been unsuccessful so far. The success in obtaining a method for high production of flavonoid might very well be one of the solutions for this problem in the future.
Culture Techniques ; Flavonoids ; biosynthesis ; Plant Roots ; growth & development ; Rhizobium ; physiology ; Saussurea ; growth & development