Objective: To study the expression and significance of VEGF-C, D2-40, and E-Cad in Human breast carcinoma. Methods: S-P immunohistochemical method was used to assess the expression of VEGF-C in 88 cases of infiltrating ductal carcinoma of the breast (with intact clinicopathologic and follow-up in-formation) and 54 cases of different intraductal hyperplastic lesions. The relationship of VEGF-C expression with D2-40 and E-Cadherin in infiltrating ductal carcinoma of the breast was analyzed. Results: VEGF-C ex-pression was higher in the invasive group than that in the dysplasia group and benign lesions. VEGF-C ex-pression in the metastatic group was significantly higher than that in the group without lymph node metasta-sis, indicating that VEGF-C plays an important role in the growth and metastasis of breast cancer. VEGF ex-pression in breast cancer group was higher than that in the dysplasia group and benign lesions. LVD count in the group with lymph node metastasis was significantly higher than that in the group without lymph node me-tastasis. The positive rate of E-Cad expression in the invasive group was significantly lower than that in the dysplasia group and benign lesions. VEGF-C expression was positively correlated with D2-40 expression. VEGF-C expression was negatively correlated with E-Cad expression. Conclusion: VEGF-C and D2-40 could be used for the detection of early lymphatic metastasis of breast cancer. VEGF-C and E-Cad can be used as important indices for the evaluation of the prognosis of breast cancer.

Objective The purpose of this article was to investigate the internal tunnel position during anterior cruciate ligament (ACL) reconstruction with single-bundle ACL. Methods MRI were performed in 10 knees form 10 volunteers at full extension and at 30°, 60°, 90°, and 120° flexion position. All the images obtained were exported into Mimics 10.01. Three-dimensional models were established with Mimics in a computer. All the mark points were confirmed on femur and tibia. The distance between the femoral mark point and tibial mark point was measured. The isometric point was determined as the change in the distance was shorter than 3mm during knee flexion-extension. Results Ten three-dimensional models were established successfully and the isometric points of A0-X, A15-X, A30-X, A45-X, B0-Y, B15-Y, B30-Y, B45-Y, C0-Z, C15-Z, C30-Z, C45-Z, and C60-Z were identified. Conclusion There was no absolute anatomical isometric point, whereas the physiological isometric point did exist. Therefore, determination of tibial point should be considered synthetically. B45-Y was recommended for tunnel position.

Objective To express mouse IFN-λ2 stably and study its biological activity. Methods Full-length of mIFN-λ2 cDNA was obtained by using RT-PCR from cells of mouse spleen stimulated by ve-sicular stomatitis virus(VSV) and then subcloned to eukaryotic expressing vector PCAGG-EGFP. The recom-binant was transfected into CHO cells. VSV * GFP-B16 system was used to measure the antivirus activity. The constructed cell line MDBK-Mxp-Luc was used to study the character of Mx1 protein induced by the mIFN-λ2. Results The recombinant pMD18-T-mIFN-λ2 was digested by two kinds of enzyme, Sac I and Xho I, to produce the fragment was of 582 bp, and of which the sequence analysis of sequence shows it was entirely consistent with the nucleotide sequences reported in GenBank. PCAGG-EGFP-mIFN-λ2 eukaryotic expressing vector was constructed successfully and expressed stably in CHO cells, and the mRNA of mIFN-λ2 was verified expressing in CHO-PCAGG-EGFP-mIFN-λ2 cell line by RT-PCR. The antivirus activity of in the supernatant secreted by the CHO-PCAGG-EGFP-mIFN-λ2 cell line was 10~4 AU/ml. The mIFN-λ2 pro-tein can could induce the expression of the antivirus protein Mx1, and the expression of Mx1 protein induced by mIFN-λ2 enhanced with time going, 9 to 12 hours achieved the peak, 24 hours vanished. Conclusion Gene cloning of mIFN-λ2 was successful. The eukaryotic expressing vector of mIFN-λ2 was constructed suc-cessfully and expressed stably in CHO cells, and its product has obvious antivirus activity in vitro. And the antivirus activity of the product was closely correlated with inducing expression of antivirus protein Mx1.

OBJECTIVE: To investigate the effects of human cerebrospinal fluid (CSF) during development phase on migration and differentiation of fetal brain neural stem cells (NSCs).METHODS: Fetal brain cells of gestational age of 16 weeks that were frozen in liquid nitrogen were obtained, resuscitated and incubated in DMEM/F12 medium containing epithium growth factor (EGF), basic fibroblast growth factor (bFGF), B27 and N2. The neurospheres cultured for 14 days were obtained. CSF was absorbed from the subarachnoid cavity and brain ventricle in the embryonic group. CSF was collected by lumbar puncture or ventricular puncture in the child group. The neurospheres cultured for 14 days were transplanted into the pure CSF in an incubator containing 5% CO_2 at 37 ℃. Cellular migration and growth of neurospheres in CSF were observed. Effects of CSF on neural cell differentiation were identified by immunofluorescence. RESULTS: Neural stem cells in the form of neurospheres derived from fetal brain were inoculated into the pure CSF, and cell migration were commonly observed besides few of neurospheres in child CSF culture at 6 hours following culture. Surrounding cells of neurospheres extended processes, forming cell cord that became cell webs after extension. Compared with the embryonic group, positive rate of glial fibrillary acidic protein was significantly increased in the children group (P < 0.01), but positive rates of nerve fiber and nestin were significantly decreased (P < 0.01). In addition, galactocerebroside-positive cells were only found in 3 baby CSF cultures. CONCLUSION: There existed significant affections on both migration and differentiation of human neural stem cells when cultured in pure CSF with different developmental phase, suggesting that CSF is one of major niche factors for central neural system development.

Objective To isolate and identify the potential binding partners of LRRK2,a gene linked to both dominant familial form and sporadic form of Parkinson's disease,thus to further our knowledge of its function.Methods We used a sequence containing full-length of COR domain and part of ROC and MAPKKK domain as bait.The bait amplified by polymerase chain reaction(PCR) was then cloned into a yeast expression plasmid pGBKT7.After being sequenced and analyzed,pGBKT7-bait was transformed into the yeast strain AH109.Western blot was performed to confirm the expression of pGBKT7-bait in AH109 yeast strain.Then human fetal brain cDNA library was trarnsformed into that yeast strain.which could express pGBKT7-bait fusion protein.The yeast strain which contained pGBKT7-bait and human fetal brain cDNA library was plated on quadruple dropout medium (SD/-Trp/-Leu/-His/-Ade)containing X-a-gal.We retested these positive colonies using 2 independent yeast strains AH109 contained pGBKT7-bait or pGBKT7,respectively.At last,these plasmids isolated from these true positive colonies were analyzed by bioinformatics.Results We obtained 9 true positive colonies,these colonies were sequenced, and we performed sequence Blast in GenBank.Three colonies of the 9 positive colonies were not in open reading-frames.Among other 6 colonies,there were known proteins including spermatid perinuclear RNA-binding protein(STRBP)and BCL2-associated athanogene 5 isoform b(BAG5),as well as unknown proteins including tyrosine phosphatase non-receptor type(PTPN23),1(3)mbt-like 3 isoform b(L3 MBTL3),RALY RNA binding protein-like isoform 1(RALYL),and Homo sapiens mRNA for KIAA1783 protein,partial cds(KIAA 1783).Conclusion True positive colonies of LRRK2 are successfully obtained by the yeast 2-hybrid.Our screened proteins may provide a new research clue for revealing biological functions of LRRK2,pathogenesis of Parkinson's disease,and other neurodegerations.

OBJECTIVETo learn about the photosynthetic characteristics of Sarcandra glabra and provide the theoretic references for its better planting.

METHODThe photosynthetic parameters of twenty different provenances of Sarcandra glabra were determined by Li-6400 portable photosynthesis system, and the data was analyzed by Excel and SAS software.

RESULTThe results showed that the light saturation point of different Provenances of S. glabra were almost about 800 micromol x m(-2) x s(-1), while the light compensation point of them were from 14.70 micromol x m(-2) x s(-1) to 48.68 micromol x m(-2) x s(-1). The curve of net photosynthetic rate had two peaks on sunny day, the first one was in the morning and the other one was in the afternoon. The photosynthetic "noon- break" of S. glabra appeared between 11:00-13:00, when the net photosynthetic rate goes down sharply. Intercellular CO2 concentration (C(i)), CO2 concentration (CO2S) and transpiration rate (T(r)) all have effect on the diurnal change of net photosynthetic rate (P(n)) of S. glabra, and the average correlation coefficient between P(n) and the parameters above were orderly as -0.89 (P < 0.01), -0.75 (P < 0.05) and 0.69 (P < 0.05);

CONCLUSIONS. glabra was a plant with characteristics of shade-tolerance, and through the way of covering, sprinkling for decreasing the surrounding temperature would be effective to reduce its "noon-break" time and increas its efficiency of photosynthesis.

Magnoliopsida ; classification ; physiology ; radiation effects ; Photosynthesis ; radiation effects ; Phylogeny ; Sunlight

OBJECTIVETo compare the pain thresholds of acupoints in the meridians related to irritable bowel syndrome(IBS) between IBS patients and healthy people.

METHODSThirty-four healthy adults were collected into a normal group,and 60 patients with IBS were arranged into an IBS group. Pain thresholds were detected on the acupoints of large intestinal,small intestinal,stomach,spleen,gallbladder meridians and some commone use acupoints for IBS by pain measuring apparatus three times. Bilateral-well,-spring,-stream,-river,-sea,front-alarm,lower-sea,-primary,-connecting and back-transport points of each meridian were selected as the main acupoints. And pain thresholds of acupoints common used in clinic were compared between the two groups.

RESULTSThe thresholds of five-transport points and-connecting point of the large intestinal meridian in the IBS group were apparently lower than those in the normal group(all<0.05),with Hegu(LI 4),Tianshu(ST 25),Shangjuxu(ST 37) and Dachangshu(BL 25) more decreasing(all<0.01). The thresholds of five-transport points,front-alarm point and-primary point of the small intestinal meridian in the IBS group were obviously lower than those in the normal group(all<0.05),with Zhizheng(SI 7),Xiajuxu(ST 39) and Xiaochangshu(BL 27) more decreasing(all<0.01). The thresholds of Zusanli(ST 36),Weishu(BL 21),Yanglingquan(GB 34),Danshu(BL 19),Pishu(BL 20),Neiguan(PC 6),Taichong(LR 3),Taixi(KI 3),Sanyinjiao(SP 6) and Qihai(CV 6) in the IBS group were markedly lower than those in the normal group(all<0.05).

CONCLUSIONSThe pathological changes of IBS often appeared in the stomach,large intestinal,small intestinal,bladder meridians,and some acupoints in the liver,spleen and kidney meridians. When the functions of viscera are abnormal,the pain thresholds of related acupoints tend to decrease,and meridians and acupoints become sensitized from quiet state.

Antibodies, Neutralizing ; Antibodies, Viral/isolation & purification* ; B-Lymphocytes/virology* ; COVID-19/virology* ; Humans ; Immunity/genetics* ; RNA, Viral/immunology* ; SARS-CoV-2/pathogenicity* ; Single-Cell Analysis