Objective To establish the normal parameters of psychometric measures such as the number connection tests A(NCT-A)and digit symbol tests(DST)in assessment of minimal hepatic en- cephalopathy(MHE).Methods One hundred and sixty healthy volunteers(aged 25 to 64 years;educa- tional level＞9 years)were divided into＜35 ys,35～44 ys,45～54 ys and 55～64 ys groups.All of the healthy volunteers were assessed with NCT-A and DST to establish the normal value of age-related parameters,which can be used for diagnosis of MHE in patients with liver cirrhosis.Two standard devi- ation of the normal mean was used as a diagnostic criterion for MHE.One hundred and six cirrhotic patients were assessed with these parameters.Results The parameters of NCT-A were(25.1?4.6) sec in＜35 ys group,(32.1?6.8) sec in 35～44 ys group,(38.6?7.1)sec in 45～54 ys group or (49.3?6.3)sec in 55～64 ys group.The scores of DST were 49.9?4.7 in＜35 ys group,44.6?4.8 in 35～44 ys group,38.5?5.0 in 45～54 ys group or 35.4?4.7 in 55～64 ys group.Thirty one out of 106 cirrhotic patients were diagnosed as MHE based on these parameters.Conclusion The NCT- A and DST are psychometric assessments for diagnosis of MHE.Age-based normal paramerters of NCT- A and DST are needed to be established.

Adult ; Electroencephalography ; Evaluation Studies as Topic ; Female ; Hepatic Encephalopathy ; diagnosis ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Tomography, X-Ray Computed

OBJECTIVETo modify the current PCR-based method for rapid and efficient preparation of small interfering RNA (siRNA) expression cassette to improve the efficiency of RNA interference.

METHODSThe U6 promoter sequence was amplified by PCR using the genomic DNA of K562 cells as the template, and cloned into pMD18-T vector which served as the template for further PCR amplification with the primers on the plasmid. The amplified product was directly used as the template for preparing siRNA expression cassette. The siRNA expression cassette targeting p53 gene was amplified, verified by sequencing, and transfected into SH-SY5Y cells. After a 48-hour transfection, the cells were harvested and the total RNA was for RT-PCR for evaluating the effect of RNA interference.

RESULTSThe sequencing result confirmed the correct U6 promoter sequence cloned from K562 cells. After transfection of SH-SY5Y cells for 48 h with siRNA expression cassette, the p53 gene expression was inhibited at the mRNA level in comparison with the control cells as demonstrated by RT-PCR detection.

CONCLUSIONThe siRNA expression cassette prepared using the established method described hereby can be well applicable in RNA interference research.

Gene Silencing ; Gene Targeting ; methods ; Genetic Vectors ; genetics ; Humans ; K562 Cells ; Polymerase Chain Reaction ; RNA, Small Interfering ; biosynthesis ; genetics ; RNA, Small Nuclear ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

Objective To explore the clinical value of the level of granzyme B and perforin mRNA in urine for the diagnosis of renal transplantation patients with delayed graft function (DGF). Methods Twenty-four cases of renal transplantation patients with DGF were included in this study. Seventy-three u-rine specimens were obtained from these patients who received graft biopsies. Among the 24 cases, ureteral obstruction occurred in 2 cases, vascular thrombosis in 1 case, acute CsA intoxication in 3 cases, acute tubu-lar necrosis (ATN) in 7 cases, ATN complicating borderline change in 2 cases, ATN complicating acute re-jection (AR) in 3 cases, AR in 6 cases. Total RNA was isolated from the urinary cells. Messenger RNA (mRNA) encoding the cytotoxic proteins perforin and granzyme B gene were measured with the quantitative polymerase-chain-reaction assay-(RT-PCR). SPSS13.0 software was used for data analysis. Levels of mRNA were log-transformed before analysis. Results The levels of perforin and granzyme B mRNA in u-rine among the ureteral obstruction group, vascular thrombosis group, acute CsA intoxication group and ATN group were very low. There was no significant difference among these groups (P>0.05). However,among the ATN complicating borderline change group 1.22, 0. 97 fg/μg, ATN complicating AR group (1.20±0.39), (1.07±0.30)fg/μg, and AR group(11.13±0. 33), (1.01±0.19)fg/μg, the levels were increased significantly(P<0.001). Conclusion Measurement of mRNA encoding the cytotoxic proteins perforin and granzyme B gene in urinary cells in renal transplantation patients with DGF could be helpful to etiological diagnosis of DGF, and might be used as an index for the appropriate management of the borderline change.

OBJECTIVETo study the correlation of homocysteine (Hcy) in plasma with nitric oxide synthetase (NOS) and endogenous carbon monoxide (CO) in the penile corpus cavernosum of type 2 diabetic rats.

METHODSThis study included 40 male Wistar rats, 10 as controls (Group A) and the other 30 as diabetes mellitus (DM) models. Four weeks after the model establishment, the model rats were divided into a DM group (Group B, n = 10), an insulin treated group (Group C, n = 10), and a folic acid and vitamin B12 treated group (Group D, n = 10). All the rats were injected with apomorphine and observed for penile erection at 8 and 12 weeks, and the levels of total plasma Hcy (tHcy), NOS and CO in the penile corpus cavernosum were measured at 12 weeks.

RESULTSCompared with Group A, the level of tHcy was significantly increased, while NOS and CO activities in the penile cavernous tis-sue and erectile function remarkably decreased in Group B (P < 0.01). The incidence rate of high Hcy was 55% in the DM rats. In comparison, the level of tHcy was obviously decreased, and the NOS activity and erectile function markedly increased in Groups C and D (P < 0.01). The Hcy level showed a significant negative correlation with NOS activity (rA = -0.89, rB = -0.76, rc = -0.91, rD = -0.91) and CO content (TA = -0.82, r, = -0.77, rc = -0.93, rD = -0.81).

CONCLUSIONHigh plasma Hcy can decrease NOS and CO activities in the penile corpus cavernosum, and consequently induce erectile dysfunction in DM rats, while insulin, folic acid and vitamin B12 can improve their penile erectile function by increasing NOS and CO activities.

Animals ; Carbon Monoxide ; metabolism ; Diabetes Mellitus, Experimental ; blood ; physiopathology ; Diabetes Mellitus, Type 2 ; blood ; physiopathology ; Folic Acid ; pharmacology ; Homocysteine ; blood ; Insulin ; pharmacology ; Male ; Nitric Oxide Synthase ; metabolism ; Penis ; drug effects ; metabolism ; Rats ; Rats, Wistar ; Vitamin B 12 ; pharmacology

OBJECTIVETo validate transient elastography (Fibroscan) in assessment of hepatic fibrosis in autoimmune hepatitis (AIH).

METHODSLiver stiffness was assessed using Fibroscan in totally 30 patients with AIH. We compared the results of Fibroscan with the Scheuer fibrosis stage in liver biopsy in each patient.

RESULTS4 patients were shown as liver fibrosis stage S0, 6 as S1, 5 as S2, 11 as S3 and 4 as S4. Failure of the Fibroscan measurement occurred in 1 case (3.3%) because of her increased body mass index (BMI). The stiffness of Fibroscan was significantly correlated with the liver biopsy fibrosis stage (r = 0.801, P less than 0.001). The liver stiffnesses between mild and moderate fibrosis (S0-2) and advanced fibrosis (S3-4) were significantly different (t = -3.937, P = 0.001).

CONCLUSIONTransient elastography (Fibroscan) is a promising non-invasive method for detection of fibrosis in patients with autoimmune hepatitis. Its use for the follow up and management of these patients and should be evaluated further.

Elasticity Imaging Techniques ; methods ; Hepatitis, Autoimmune ; diagnostic imaging ; Humans ; Liver ; diagnostic imaging ; Liver Cirrhosis ; diagnostic imaging

The automatic cleaning machine we have developed, adopts a SCM system in automatic cleaning. The machine has five functions: ultrasonic cleaning, cold or hot water spraying, drying and greasing. The clinical applications show that the machine with a good effectiveness is suitable for the cleaning of many surgical instruments. It also raises working efficiency, cuts down on the cost of repair and maintenance and reduces the injury and infection to nurses caused by manual cleaning, satisfying the needs of clinical applications.
Automation ; instrumentation ; Disinfection ; instrumentation ; Equipment Design ; Surgical Instruments ; standards ; Ultrasonics ; instrumentation

OBJECTIVETo observe the effect of Tongsaimai (TSM) tablets in treating foot trauma of diabetic foot (DF) model rats, and discuss its potential mechanism.

METHODMale SD rats were selected to duplicate the diabetic foot ulcer model and randomly divided into the blank control group, the model group, the metformin treatment group, and TSM 12.44, 6.22, 3.11 g x kg(-1) groups (n = 10). The healing of ulcer wounds were observed on day 1, 4, 8, 13 and 18. After 18 days, a histopathologic examination was conducted for ulcer tissues. The contents of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by hydroxylamine and TBA methods. The content of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were determined with the radioimmunoassay. The immunohistochemical method was used to observe the expression of vascular endothelial growth factor (VEGF) in ulcer tissues and the number of capillary vessels.

RESULTTSM could alleviate the pathological changes of diabetic foot rats, accelerate the ulcer healing on 4, 8, 13, 18 d, reduce MDA, IL-6, TNF-alpha, VEGF content in rat serum at 18 d (after the rehabilitation period), and enhance the SOD content. Specifically, the TSM 12.44 g x kg(-1) group showed significant differences compared with the model group (P < 0.05, P < 0.01). At 18 d after the treatment (the late rehabilitation period), the VEGF expression of TSM 12.44, 6.22 g x kg(-1) groups and the number of blood capillaries of the TSM 12.44 g x kg(-1) group were significantly lower than that of the model group (P < 0.05, P < 0.01).

CONCLUSIONTSM could promote the foot wound healing of DF model rats, reduce MDA, IL-6 and TNF-alpha levels in serum, increase the SOD content and decrease the VEGF expression and the number of blood capillaries in the late rehabilitation period. Its action mechanism may be related to the inhibition of oxidative stress injury and the inflammatory cell infiltration.

Animals ; Diabetic Foot ; drug therapy ; genetics ; metabolism ; physiopathology ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Interleukin-6 ; genetics ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; genetics ; metabolism ; Tablets ; administration & dosage ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Wound Healing ; drug effects

Objective To investigate the significance of fibroblast growth factor 23 in metabolic bone disease of prematuriy.Methods 60 patients who had been treated in our hospital from March 2016 to March 2017 were included in this study.Blood biochemistry was examined two weeks after birth,and values of blood phosphorus, serum calcium and alkaline phosphatase were recorded. Serum levels of 25 hydroxyvitamin D3,parathyroid hormone and fibroblast growth factor 23 were detected two weeks after birth. 20 premature infants with metabolic bone disease were selected as a study group. 40 infants without metabolic bone disease were treated as a control group. Two weeks after treatment,the above indicators were measured and compared in the study group. Results Serum levels of 25 hydroxyvitamin D3,parathyroid hormone and fibroblast growth factor 23 were compared between the two groups 2 weeks after birth,the difference was statistically significant(P<0.05).Levels of serum parathyroid hormone and fibroblast growth factor 23 in the study group were not statistically significant after treat-ment(P > 0.05). Levels of 25 hydroxy vitamin D3 in the study group had statistically significant after treatment (P<0.05).Conclusions Early detection of fibroblast growth factor 23 can reflect metabolic bone disease in pre-term infants.It suggests that vitamin D should be adequately supplemented in early.

OBJECTIVETo create a standard mini-swine model of chronic ischemic myocardium by endoscopy for the research of gene transfer and stem cell.

METHODSTwenty-three male China experimental minipigs were used, aged from 8 to 11 months with a mean of (9.3 +/- 1.8) months and weighed from 20 to 30 kg with a mean of (29.3 +/- 4.3) kg. The myocardial ischemia was established by gradual occlusion of the left circumflex coronary artery (LCX) with an Ameroid constrictor. The Ameroid constrictor was implanted around LCX by endoscopy. Selective coronary angiography, electrocardiogram and Echo-Doppler study were performed perioperatively to evaluate the degree of stenosis.

RESULTSChronic ischemic myocardial models were successfully generated in 20 of 23 swine by full-endoscopy. Ameroid constrictors were placed at the LCX accurately. Three swine died of anesthetic accident, cardiac arrhythmia at secondary coronary angiography, and pulmonary infection within 6 weeks after operation respectively. Operation time was 25 to 65 min with a mean of (46 +/- 9) min. The blood loss was 30 to 60 ml with a mean of (55 +/- 12) ml. Six weeks later, coronary angiography revealed the total occlusion and partial stenosis (> 85%) of the LCX occurred in 7 and 13 swine respectively. Cardiac systolic and diastolic dysfunction were found in all swine. The ejection fraction value was (65.0 +/- 6.3)% before operation and (41.0 +/- 9.3)% after operation (P = 0.008). The fractional shortening value was (36.2 +/- 4.3)% before operation and (34.2 +/- 2.3)% after operation (P = 0.027).

CONCLUSIONThe endoscopic surgery is a less invasive way to create a standard mini-swine model of chronic ischemic myocardium with effective results.

Animals ; Disease Models, Animal ; Feasibility Studies ; Male ; Myocardial Ischemia ; Swine ; Swine, Miniature ; Thoracoscopes