NS2 and NS3 are two post-transcriptional gene silencing suppressors that are encoded by Rice stripe virus. Gene silencing suppressors are always related to the pathogenicity of viruses. In this study, the cDNA of NS2 and NS3 were recombined by overlapping PCR assays, ligated to the RNAi vector, and inserted into the PXQ expression vector using Pst I; the expressed vector was transferred into calluses induced from seeds of the japonica rice cultivar, 'Nipponbare', using an Agrobacterium-mediated method. Thirty-one T0 transgenic plants were selected by G418 screening. PCR and southern blot analyses confirmed that the target gene was transformed into transgenic rice successfully, and different transgenic plants contained various copies of the gene. The disease resistance assay revealed that T0 transgenic rice had a delayed onset of RSV for approximately 10-20 d, and the accumulation of virus in the transgenic plants was reduced by 30%-50%. This was related to the delayed onset of disease.
Disease Resistance ; Oryza ; genetics ; immunology ; virology ; Plant Diseases ; genetics ; immunology ; virology ; Plants, Genetically Modified ; genetics ; immunology ; virology ; RNA Interference ; Tenuivirus ; genetics ; immunology ; Viral Nonstructural Proteins ; genetics ; immunology

Objective To observe the clinical efficacy of scalp cluster needling in treating essential tremor.Method Sixty patients with essential tremor were randomized into two groups, 30 cases each. The treatment group was intervened by scalp cluster needling plus acupuncture at the bilateral dancing tremor control zone; the control group was intervened by acupuncture at the dancing tremor control zone alone. The total effective rate and tremor score were observed after 30-day treatment, and the relapse rate was investigated 3 months after the intervention via telephone. Result The total effective rate was 96.7% in the treatment group, significantly higher than 73.3% in the control group (P<0.05); the tremor score of the treatment group was significantly lower than that of the control group at the end of the intervention (P<0.01); the 3-month telephone follow-up study showed that the relapse rate of the treatment group was significantly lower than that of the control group (P<0.05).Conclusion Scalp cluster needling plus acupuncture at bilateral dancing tremor control zone can produce a more notable efficacy in treating essential tremor compared to acupuncture at the dancing tremor control zone alone.

ABSTRACT The taboos of man-spirit (RenShen) and bottom-spirit (KaoShen) are two kinds of time-related contraindications that are more commonly seen in the literature of acupuncture and moxibustion. The meaning of man-spirit is often extended to qi-blood or mind, or else directly equals to heaven-spirit (TianShen). It is highly possible that the bottom-spirit is subordinate to man-spirit, and both Taoist philosophy and the SanCai theory declare the importance of bottom in human body. The violation of contraindications could lead to diseases in the neighborhood where man-spirit travels. However, the results are generally recorded as ulcer, lingering disease and death in most medical texts. From Ming dynasty, more and more doctors have expressed disapproval of man-spirit and bottom-spirit.
Acupuncture Therapy ; psychology ; Humans ; Moxibustion ; psychology

Objective To investigate the expression of Mta-1 in bladder transitional cell carcinoma (BTCC)and to analyze its correlation with the clinical staging,pathologic grading,metastasis and recur- rence,and to explore the possible molecular mechanisms.Methods Samples of 42 cases of BTCC and 12 normal bladder mueosa tissues were examined with immunohistochemical analysis for the expression of Mta- 1,ER,u-PA and PAI-1.Endothelial cells were stained by anti-CD34,and microvascular density(MVD)of carcinoma tissue was calculated.The correlation of Mta-1 expression with the invasion,metastasis,angiogene- sis and recurrence of BTCCs was analyzed;and the correlation of Mta-1 expression with ER,u-PA,PAI-1,and MVD was also analyzed.Results The positive rate of Mta-1 expression in BTCCs was 73.8%(31/42) , while it was 0.0% in normal bladder mucosa tissues(P＜0.01).The expression level of Mta-l increased with the higher clinical stages and pathologic grades of BTCCs;it was higher in recurrence group(100.0% , 15/15)than in non-recurrence group(59.3%,16/27),and high in metastasis group(100.0%,14/14) than in non-metastasis group(60.7%,17/28)(P＜0.05).The expression level of ER increased with the lower clinical stages and pathologic grades of BTCCs;the positive rate of ER expression was 0.0% in 14 ca- ses with metastasis and was 53.6% in 13 of 28 cases without metastasis(P＜0.05);and the rate was 6.7% in 1 of 15 cases with recurrence and 44.4% in 12 of 27 cases without recurrence(P＜0.05).Negative cor- relation was found between Mta-1 and ER expression(r=-0.739,P＜0.01).The positive rate of u-PA ex- pression(59.5%,25/42)was significantly higher in BTCCs than that in normal bladder mucosa tissues (16.7%,2/12)(P＜0.05).Positive correlation was found between u-PA and Mta-1 expression(r= 0.875),while negative correlation was found between u-PA and PAI-1 expression(r=-0.535).The posi- tive rate of PAI-1 expression in normal bladder mucosa tissues(50.0%,6/12)was significantly higher than that in BTCCs(19.0%,8/42)(P＜0.05).In addition,negative correlation was found between PAI-1 and Mta-1 expression(r=-0.706).And positive correlation was found between MVD in BTCCs marked by an- ti-CD34 and Mta-1 expression(r=0.683).Conclusions Mta-1 is highly expressed in BTCCs,and it correlates closely with tumor pathologic grades,clinical stages,recurrence and metastasis.Mta-1 up-regulates the expression of u-PA and down-regulates that of PAI-1,which is associated with invasion and metastasis and acts as an angiogenic mediator in BTCCs.A negative correlation is found between Mta-1 and ER in inva- sion and metastasis of BTCCs.

Objective To explore the impact factors for post-thaw embryo survival rate and clinical pregnancy rate in frozen-thawed embryo transfer program. Methods The clinical data of 573 cycles of frozen-thawed embryo transfers were retrospectively analysed. Groups were divided according to the pre-freeze embryo quality, pre-freeze embryonic developmental stage, frozen-thawed embryo quality and cryopreservation technique, respectively, and post-thaw embryo survival rates and/or clinical pregnancy rates were compared among groups. Results The clinical pregnancy rate of high quality pre-freeze embryo was significantly higher than that of low quality pre-freeze embryo (31.8% vs 20.0%) (P< 0.05). There was no significant difference in the post-thaw survival rates and clinical pregnancy rates between embryos frozen at day 2 of ferrtilization and those frozen at day 3 of ferrtilization(79. 1% vs 82.9% and 25.5% vs 31.2%, respectively) (P>0.05). The clinical pregnancy rates of the transfer cycles only with fully intact embryos and with mixed embryos were significantly higher than that only with partially damaged embryos(36.7% vs 24.1% and 29.2% vs 24.1%, respectively)(P<0.05). The post-thaw survival rate and post-thaw high-quality embryo rate were significantly higher in those processed with modified cryopreservation technique than in those processed with original cryopreservation technique (82.0% vs 66.3% and 50.0% vs 27.5%, respectively)(P<0.05). Conclusion Pre-freeze embryo quality, post-thaw embryo survival rate and post-thaw embryo quality have a positive correlation to subsequent clinical pregnancy rate. Favorable cryopreservation technique may ensure the success of post-thaw embryo recovery and transfer.

OBJECTIVETo search for a new method of screening for molecular targets for androgen-dependent prostate cancer.

METHODSWe collected tissue samples and paired serum samples from 3 cases of androgen-dependent prostate cancer (ADPC) treated by surgical resection, and included another 3 samples of benign prostatic hyperplasia (BPH) tissue and normal human serum in the control group. The total proteins extracted were separated and transmembrane by two-dimensional gel electrophoresis, followed by hybridization with the sera of the patients with ADPC and those with hormone-independent prostate cancer (HIPC) as the primary antibodies. The differentially expressed proteins were compared by Western blot, analyzed by MALDI-TOF-MS mass spectrography, and verified by RT-PCR and Western blot following bioinformatic identification.

RESULTSThis modified method exhibited a significantly better effect in displaying differentially expressed proteins, by which 12 differentially expressed protein spots were identified, including Beclin1, glutathione S-transferase P (GSTP1-1), ZBTB7, dihydrodiol dehydrogenase 2 (DDH), enolase (ENO1), glucose-dependent insulin-releasing peptide receptor (GIPR), Mn-superoxide dismutase (MnSOD), phosphoglycerate mutase 1 (PGAM1), amino-peptidyl-prolyl cistrons isomerase (PPIA), and phospholipid-PE-binding protein (PEBP). The mRNA and protein expressions of Beclin1 were significantly down-regulated in androgen-dependent prostate cancer tissues.

CONCLUSIONThis modified serum-guided immunoblotting technique has provided a new method for clarifying the molecular mechanisms of the occurrence and progression of HIPC, in which Beclin1-mediated autophagy may play a key role.

Biomarkers, Tumor ; blood ; Blotting, Western ; Humans ; Immunoblotting ; methods ; Male ; Mass Spectrometry ; Prostatic Neoplasms ; genetics ; metabolism ; Proteomics

OBJECTIVETo establish immortalized cell line from the urothelium of the urinary bladder and identify the characteristics of the cell line.

METHODSHuman papillomavirus 16 (HPV-16) plasmid was used to transfect urothelium of infant urinary bladder in vitro with the help of Fugene-6, and this plasmid contained E6 and E7 genes of HPV-16. We also identified the existence of HPV-16 E6 and E7 genes and the biological characteristics of the cell line by PCR, immunohistochemistry, and the biology identification.

RESULTSBLTR-4 cell line, produced from the transfection of HPV-16K plasmid, was a cell line from urothelium with the expression of HPV-16 E6 and E7 genes. It had been cultured more than 70 passages, and the characteristics of growth was similar to the immortalized cell line as reported.

CONCLUSIONSBLTR-4 cell line is an immortalized cell line from urothelium of the urinary bladder, which contains HPV-16 E6 and E7 genes. BLTR-4 cell line is a good experimental model to investigate the relationship of the infection of high risk HPV and transitional cell carcinoma (TCC) in vitro.

Cell Line, Transformed ; Humans ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; Papillomavirus E7 Proteins ; Papillomavirus Infections ; virology ; Plasmids ; genetics ; Repressor Proteins ; genetics ; Transcription, Genetic ; Transfection ; Tumor Virus Infections ; virology ; Urinary Bladder ; cytology ; Urinary Bladder Neoplasms ; virology

Objective To observe the clinical efficacy of scalp cluster needling plus acupuncture at Conception Vessel points in treating generalized anxiety disorder.Method Sixty eligible subjects were divided into a treatment group and a control group by the random number table, 30 cases each. The treatment group was intervened by scalp cluster needling plus acupuncture at Conception Vessel points including Tiantu (CV22), Danzhong (CV17), Zhongwan (CV12) and Xiawan (CV10); in the control group, Shenmen (HT7), Daling (PC7), Neiguan (PC6), Qimen (LR14), Xinshu (BL15), Hegu (LI4) and Taichong (LR3) were selected. The treatments were given once a day, 6 sessions followed by 1-day interval as a treatment course, for successive 8 courses in total. The Hamilton Anxiety Scale (HAMA) scores of the two groups were compared before and after the treatment and in the follow-up study; the clinical efficacies were also compared.Result The treatment protocols in the two groups were both effective in treating generalized anxiety disorder, while the markedly effective rate was significantly higher in the treatment group than in the control group (P<0.05). The HAMA scores after treatment and in the 1-month follow-up study were significantly different from the score before the treatment in both groups (P<0.01). There were significant differences in comparing the HAMA score between the two groups respectively after treatment and in the 1-month follow-up study (P<0.05). Conclusion Scalp cluster needling plus acupuncture at Conception Vessel points shows significant advantage in treating generalized anxiety disorder compared to the ordinary acupuncture method.

Objective To investigate the number and distribution of un-repaired cleft lip/palate patients in Gansu province. Methods A census was conducted by staff members of the health system and Population and Family Planning Committee (PFPC) in Gansu province. Standardized -Questionnaires were administered to collect demographic, domestic and diagnosis-related information. Results In 2008, the total number of un-repaired cleft lip/palate patients was 4675,with a detection rate of 1.84/10 000 in Gansu province. Rates of detection were higher in males (2.11/10 000), young age group(4.86/10 000), rural areas(2.23/10 000),poor counties(2.19/10 000)than in females(1.43/10 000), medium (0,97/10 000)/old age group(0.68/10 000), township areas (0.62/10 000), or richer counties (1.35/10 000). Among all the cities and prefectures of Gansu,Baiyin city (2.7/10 000) had the highest while Jinchang city (0.7/10 000) had the lowest prevalence rates. Conclusion Un-repaired cleft lip/palate had been a disease burden to Gansu province,especially in the rural area and poorer counties.

OBJECTIVE: To evaluate the therapeutic effect and safety of holmium:YAG laser lithotripsy for treating ureteral calculi combined with acute renal failure. METHODS: Ureteroscopic holmium: YAG laser lithotripsy was used in 13 cases of ureteral calculi. RESULTS: After the operation the serum Bun and Cr levels in the patients decreased to different degrees or returned to normal and the stone free rate was up to 92.3% (12/13). CONCLUSION The ureteroscopic holmium:YAG laser lithotripsy can be the first choice for the upper urinary tract obstruction associated with acute renal failure,due to its safety and efficiency. It can also deal with the double sites of ureteral diseases.
Acute Kidney Injury ; etiology ; therapy ; Adult ; Aged ; Female ; Holmium ; Humans ; Lithotripsy, Laser ; methods ; Male ; Middle Aged ; Ureteral Calculi ; complications ; Ureteroscopy