To knock out β-lactoglobulin (BLG) gene and insert human lactoferrin (hLF) coding sequence at BLG locus of goat, the transcription activator-like effector nucleases (TALEN) mediated recombination was used to edit the BLG gene of goat fetal fibroblast, then as donor cells for somatic cell nuclear transfer. We designed a pair of specific plasmid TALEN-3-L/R for goat BLG exon III recognition sites, and BLC14-TK vector containing a negative selection gene HSV-TK, was used for the knock in of hLF gene. TALENs plasmids were transfected into the goat fetal fibroblast cells, and the cells were screened three days by 2 μg/mL puromycin. DNA cleavage activities of cells were verified by PCR amplification and DNA production sequencing. Then, targeting vector BLC14-TK and plasmids TALEN-3-L/R were co-transfected into goat fetal fibroblasts, both 700 μg/mL G418 and 2 μg/mL GCV were simultaneously used to screen G418-resistant cells. Detections of integration and recombination were implemented to obtain cells with hLF gene site-specific integration. We chose targeting cells as donor cells for somatic cell nuclear transfer. The mutagenicity of TALEN-3-L/R was between 25% and 30%. A total of 335 reconstructed embryos with 6 BLG-/hLF+ targeting cell lines were transferred into 16 recipient goats. There were 9 pregnancies confirmed by ultrasound on day 30 to 35 (pregnancy rate of 39.1%), and one of 50-day-old fetus with BLG-/hLF+ was achieved. These results provide the basis for hLF gene knock-in at BLG locus of goat and cultivating transgenic goat of low allergens and rich hLF in the milk.
Animals ; Animals, Genetically Modified ; genetics ; Female ; Fibroblasts ; Gene Knock-In Techniques ; Gene Knockout Techniques ; Goats ; genetics ; Humans ; Lactoferrin ; genetics ; Lactoglobulins ; genetics ; Milk ; chemistry ; Nuclear Transfer Techniques ; Plasmids ; Pregnancy ; Transfection

An experiment was conducted using cultivated Glycyrrhiza uralensis in age of one year to study the effects of abscisic acid (ABA) on chemical components content and color of G. uralensis. By using different concentrations of ABA spraying on leaves, the change of the chemical component content was analyzed within 45 d after ABA stimulation, and the effects on quality were studied combined with colorimetric analysis data. It turned out that in some sense the content of glycyrrhizic acid and liquiritin had increased within 45 d, especially for liquiritin. After high concentrations of ABA (3.96 mg · L(-1)) stimulating, the content of glycyrrhizic acid rose 52% while liquiritin up 392% within 30 d. Then they both showed a decline in the content of glycyrrhizic acid and liquiritin on 45 d. Color index values of a* and b* were all significantly higher than that of the control group within 45 d, which meant the color of powders turned toward red and yellow. The conclusion was that ABA (3.96 mg · L(-1)) stimulating could not only improve the quality in the traditional sense through the color of G. uralensis, but also in the modern sense by improving the content of glycyrrhizic acid and liquiritin.
Abscisic Acid ; pharmacology ; Color ; Drugs, Chinese Herbal ; chemistry ; Flavanones ; analysis ; Glucosides ; analysis ; Glycyrrhiza uralensis ; chemistry ; drug effects ; growth & development ; Glycyrrhizic Acid ; analysis ; Plant Growth Regulators ; pharmacology

OBJECTIVETo evaluate the effect of integrative medical treatment (IMT) with serial Chinese recipes on quality of life (QOF) of rehabilitation stage in SARS patients.

METHODSEighty-five SARS patients of rehabilitation stage were enrolled in the clinical study. They were divided into the IMT group (62 patients received serial Chinese recipes and western medicine) and the control group (23 patients received western medicine alone). The serial Chinese recipes were given according to patients' syndrome, one dose per day for oral intake for 3 weeks. QOF scoring in patients was observed.

RESULTSQOF scoring in the IMT group before treatment was not significantly different from that in the control group. After 3 weeks treatment, it improved to some extent in both groups, but the improvement in the IMT group was superior to that in the control group in respect of total score and score of psychologic emotional factors.

CONCLUSIONSerial Chinese recipe could improve QOF of rehabilitation stage in SARS patients.

Adolescent ; Adult ; Aged ; Convalescence ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Ofloxacin ; therapeutic use ; Phytotherapy ; Quality of Life ; Ribavirin ; therapeutic use ; Severe Acute Respiratory Syndrome ; drug therapy ; immunology ; rehabilitation

OBJECTIVETo evaluate the application of shear wave elastography (SWE) combined with transition zone biopsy in the detection of prostate cancer (PCa).

METHODSA total of 489 patients with suspected PCa underwent transrectal ultrasonography (TRUS) and SWE-guided prostatic biopsy. We evaluated the role of SWE combined with transition zone biopsy in promoting the detection rate in comparison with the results of biopsy pathology.

RESULTSThe pathological results confirmed 221 malignant and 268 benign cases. Based on systematic biopsy, SWE combined with transition zone biopsy achieved a detection rate of 45. 19% , significantly higher than that of systematic biopsy alone (33.13%) (P < 0.05). The diagnostic sensitivity, specificity, and accuracy of SWE were significantly better than those of TRUS (P < 0.05). The mean elasticity (Emean) of SWE was remarkably higher for malignant than for benign lesions ([40.1 ± 9.5] vs [21.6 ± 8.3] kPa, P < 0.05). With 28.5 kPa as the threshold of the Emean value, the area under the ROC curve was 0. 899, and the diagnostic sensitivity and specificity were 88.71% and 86.23%, respectively.

CONCLUSIONSWE combined with transition zone biopsy could significantly improve the detection rate of prostate cancer.

Elasticity Imaging Techniques ; methods ; Humans ; Image-Guided Biopsy ; methods ; Male ; Prostate ; pathology ; Prostatic Neoplasms ; diagnosis ; diagnostic imaging ; pathology ; ROC Curve ; Sensitivity and Specificity

OBJECTIVETo research the preparation technology and dissolution of Blumea volatile oil suppository.

METHODIn order to establish the content determination and methodology inspection method of Blumea volatile oil plug, the extraction process of Blumea volatile oil was optimized by using orthogonal test. Optimization on the investigation to the suppository matrix by melting time, appearance and dissolution was carried on. The best prescription craft was determined by determining the best molding temperature, dosage of the matrix and complementary makings. The determination method of dissolution was established by investigating different dissolution method and its impact on the preparation of dissolution.

RESULTThe best conditions of steam distillation extracted Blumea volatile oil was as followed, the ratio of gardenia to liquor 1:6, 2.5% drug amount of sodium, 8 hours of extracting time. The optimum temperature for mold was 60-65 degrees C. Preparation technique of Blumea volatile oil suppository was stable, which after 45 minutes and 3 h in pH 4.5 PBS released at least 70% and 90%.

CONCLUSIONBlumea volatile oil suppository with rational prescription, simple preparation and good stability.

Asteraceae ; chemistry ; Chemistry, Pharmaceutical ; methods ; Distillation ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Oils, Volatile ; chemistry ; isolation & purification ; Plant Oils ; chemistry ; isolation & purification ; Solubility ; Temperature

Objective: To compare hemodynamic effects of infusion of diethylamine/nitric oxide (DEA/NO)and sodium nitroprusside(SNP)during controlled hypotension. Method: General anesthesia was induced in dogs. The twelve healthy adult dogs were randomly assigned into one of two groups. 0.00l％ DEA/NO or 0.01％ SNP was infused to induce mean arterial pressure(MAP) to decrease to 60％ of baseline and be maintained for 30 min. During hypotension, MAP,central venous pressure (CVP), mean pulmonary artery pressure (MPAP)and cardiac output (CO) were measured and recorded. Arterial blood gas, blood lactate and urine output were also measured. Result: HR,CVP and CO were unchanged during hypotension period in both groups, MPAP decreased correspondently with MAP. There were no significantly changes in arterial blood gas,blood lactate and urine output in both groups during hypotension. Conclusion: Controlled hypotension achieved with infusion of DEA/NO has a rapid onset and short duration of action.The hemodynamics were similar to those of SNP.

Background The visual system of animal have to optimally adjust in various environmental conditions in order to obtain stable and effective visual funetion.However,the color vision system of animals which encounter uncertainty of spectral signals should be plastic.Whether the densities of various cones and expression of opsins change with long-time spectral deprivation is unclear.Objective This study was to investigate the changes of cone density as well as the expression of corresponding opsin and mRNA following the long-term illumination of monochromatic light.Methods Thirty 3-day-old guinea pigs were randomized into 3 groups and exposed tO the 530 nm green light,400 nm purple light and white light for consecutive 8 weeks respectively.The flat-mounted retinal sample was prepared and divided into dorsal zone,ventral zone and mixed zone anatomically according to the distribution of difierent light-sensitive cone.The changes in density of cone cells sensitivited to different colored light were detected by single-1abel or double-label immunocytochemistry.The levels of opsin and its mRNA were determined using Western-blot and real-time PCR respectively.Results The density of green-sensitivity cones was significantly different in the dorsal zone of retina among green light group,purple light group and white light group (F=234.28,P<0.01).Compared with white light group,the density of green-sensitive cones in dorsal retina of green light group was obviously higher but that of purple light group wag evidently lower(q=389.68,P<0.01；q=67.11,P<0.01).No significant difference was found in the density of purple-sensitive cones in the ventral zone of retina among green light group,purple light group and white light group(F=3.14,P>0.05).The density of coexpression of the mixed cone cells was increased in green light group(q=157.55,P<0.01)but decreased in purple light roup (q=254.85,P<0.01)in comparison with white light group.The expression levels of green-opsin and green-opsin mRNA in green light group was significantly elevated(q=184.45,P<0.01；q=4.71,P<0.05),but those of purple light group were evidently declined(q=5.87,P<0.05；q=346.66,P<0.01)in comparison with white light group.There was no statistically significant differences were found in the expression of purple-opsin and its mRNA among all the groups(F=1.24,P>0.05；F=3.27,P>0.05).Conclusion After the exposure of long-time monochromatic light illumination,monochromatic cones density and its opsin in guinea pig occur the corresponding alteration to gain good spatial vision as a compensatory reaction.These outcomes imply that there is some plasticity during the development of color vision.The increase of green-sensitive cones might be from the differentiation of coexpression cones in transition region.

OBJECTIVETo evaluate surgical strategy and clinical outcomes for the treatment of thoracolumbar metastatic tumor.

METHODSFrom January 2009 to December 2010,42 patients with thoracolumbar metastatic tumor were treated surgically. Among the patients, 30 patients were male, and 12 patients were female, ranging in age from 28 to 76 years old, with an average age of 56.8 years old. Twenty-five patients had metastatic tumor in thoracic vertebraes, and 17 patients had metastatic tumor in lumbar vertebraes. Thirty-four patients had metastatic tumor in 1 segment, 6 patients had metastatic tumor in 2 segments and 2 patients had metastatic tumor in 3 segments. Two patients had no symptoms and 40 patients had back or leg pain. Eighteen patients had neurologic deficits, and 5 patients had injuries of A degree, 3 patients had injuries of B degree, 4 patients had injuries of C degree, 6 patients had injuries of D degree according to ASIA grading system. The operation goal was made according to Tomita evaluation. The surgical procedures included pallative decompression, tumor curettage and total vertebrectomy, which were decided based on Tomita classification. The pain, spinal cord function,part control of tumor,survival rate and conditions of internal fixation were evaluated at 1 week, 3 months, 6 months, 1 year and 2 years after operation.

RESULTSOne patient died in the operation. Pain relief was obtained in 38 patients after operation. Among 18 patients suffering from spinal cord compromise, 17 patients improved 1 to 4 grades after surgery according to the ASIA grading system. All the patients were followed up and the duration ranged from 24 to 48 months, with a mean time of 34.2 months. Five patients got recurrence. The postoperative survival rates at 3 months, 6 months, 1 year and 2 years were 95.2%, 85.7%, 58.2%, 37.6% respectively.

CONCLUSIONAccording to Tomita system, the different surgical treatments can be selected for patients with spinal metastatic tumors, which can relieve pain, improve the neurological status and spine stabilization, maintain local control, improve quality of life.

Adult ; Aged ; Female ; Humans ; Lumbar Vertebrae ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Spinal Neoplasms ; diagnosis ; secondary ; surgery ; Thoracic Vertebrae ; Treatment Outcome

OBJECTIVETo improve the diagnosis and treatment of priapism.

METHODSWe analyzed the types, causes, treatment and prognosis of 15 cases of priapism. The patients ranged in age from 20 to 66 (mean 46) years, their erection lasting 10-172 (mean 28.4) hours. Among them, 6 cases resulted from in taking vaso-active agents, 1 had transitional cell carcinoma of the bladder metastasized to the penis, 2 had leukemia, 3 had a traumatic history and the other 3 had unknown causes.

RESULTSOf the total number, 12 fell into the ischemic low-flow type and 3 the non-ischemic high-flow type. Follow-up lasted 1-26 months. In the 12 cases of the ischemic low-flow type, 7 were cured by 2-6 mg metaraminol injection at the root of the cavernous body and, when necessary, the perfusion of heparinized saline at the glans and the root of the cavernous body of the penis, and 2 achieved detumescence after glandular cavernosal shunting. The 2 cases caused by leukemia and 1 by metastasis of transitional cell carcinoma underwent penectomy, but with unfavorable prognosis. Of the 3 high-flow type cases, 1 was cured by selective embolization of the internal pudendal artery and the other 2 discharged after conservative treatment, but developed ED on follow-up.

CONCLUSIONCavernous blood gas analysis and color duplex ultrasonography are helpful to the accurate and timely diagnosis of priapism. Cavernosal decompression and intracavernosal injection of aramine can be applied to most of the patients. If conservative treatment fails to achieve detumescence of the penis, surgery should be performed immediately for both types of priapism.

Adult ; Aged ; Blood Gas Analysis ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Penis ; chemistry ; Priapism ; diagnosis ; etiology ; surgery ; Prognosis ; Ultrasonography, Doppler, Duplex

OBJECTIVETo improve the diagnosis and treatment of coincident vesical transitional cell carcinoma (VTCC) and prostate cancer.

METHODSWe analyzed the clinical data of 5 cases of coincident VTCC and prostate cancer.

RESULTSThe 5 patients, at the mean age of 66.2 years, were diagnosed as having grade II - III VTCC by cystoscopy and biopsy, 1 with a history of prostate cancer, and the other 4 with prostate cancer confirmed by postoperative pathological examination. Two of the patients were treated by radical cystoprostatectomy, 1 by radical cystoprostatectomy and ileum conduit surgery, 1 by transurethral resection of bladder tumor, and the other 1 by palliative ureterocutaneostomy due to cardiopulmonary problems. The follow-up lasted 8 -26 months. One of them died of diffused metastasis 20 months after surgery, 1 survived with the tumor untreated, and the other 3 remained tumor free.

CONCLUSIONCoincident VTCC and prostate cancer is easy to be missed in diagnosis. PSA detection, rectal palpation, transrectal ultrasonography, biopsy, and cystoscopy are the main diagnostic options for this disease. Its treatment should be based on the classification and clinical staging of the two cancers. Coincident VTCC and prostate cancer does not suggest poor prognosis.

Aged ; Carcinoma, Transitional Cell ; diagnosis ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Prostatic Neoplasms ; diagnosis ; pathology ; surgery ; Urinary Bladder Neoplasms ; diagnosis ; pathology ; surgery