[Dumai moxibustion combined with low-dose tadalafil for erectile dysfunction: Evaluation of therapeutic effect].

OBJECTIVE: To investigate the clinical effect of dumai (governor meridian) moxibustion combined with low-dose tadalafil in the treatment of ED with decline of vital gate fire. METHODS: We enrolled in this study 130 ED patients with decline of vital gate fire who met the inclusion criteria and equally randomized them into a control and an experimental group, the former treated with low-dose tadalafil tablets at 5 mg once a day while the latter by dumai moxibustion once a week in addition, all for 4 weeks. Of the total number of subjects, 62 in the control group and 63 in the experimental group completed the experiment. We recorded the scores on IIEF-5, Erection Quality Scale (EQS), Erection Hardness Scale (EHS), TCM symptoms and Treatment Satisfaction Scale (TSS) as well as the penile hemodynamic parameters peak systolic velocity (PSV), end diastolic velocity (EDV) and resistance index (RI) before and after treatment and compared them between the two groups. RESULTS: The total response rate was significantly higher in the experimental group than in the control (87.30% vs 66.13%, P < 0.05). IIEF-5, EQS, EHS and TSS scores, PSV and RI were markedly increased while TCM symptoms and EDV remarkably decreased in both groups after treatment (P < 0.05), even more significantly in the experimental than in the control group (P < 0.05). CONCLUSION: Dumai moxibustion combined with low-dose tadalafil can improve erectile function, increase penile blood flow velocity and alleviate clinical symptoms in ED patients with decline of vital gate fire, with definite clinical effect and safety.

Risks and predicting factors of bleeding complications in hepatitis B virus-related acute-on-chronic liver failure.

BACKGROUND/AIMS: This study aimed to provide supporting evidence for prevention and prognostic evaluation of bleeding complications in the early stage by exploring the risk and predicting factors in patients with acute-on-chronic liver failure (ACLF). MATERIALS AND METHODS: A total of 101 hospitalized patients with ACLF were retrospectively included from January 1, 2014 to December 31, 2015. The patients were divided into bleeding (n=38) and nonbleeding groups (n=63). Demographic data and laboratory tests were recorded and compared between the two groups. The incidence, risk factors, and prognosis of bleeding complications among patients with ACLF were investigated. RESULTS: A total of 38 cases (37.62%) had bleeding complications: 26 (25.74%) were spontaneous and 12 (11.88%) were postprocedural. Patients with bleeding complications had lower platelet (p=0.008), fibrinogen (p<0.001), factor V (p=0.001), and factor VII (p=0.026) levels; higher serum creatinine levels (p=0.004); and a higher proportion of cirrhosis (p=0.013). Logistic regression analysis showed that cirrhosis (odds ratio=3.251, p=0.046), fibrinogen level (odds ratio=0.352, p=0.007), and factor VII level (odds ratio=0.951, p=0.011) contributed to the development of bleeding complications. A subgroup analysis of invasive manipulation-induced bleeding complications showed lower levels of factors V (p=0.018) and VII (p=0.021) in the postprocedural bleeding group. Follow-up studies showed that the nonbleeding group had a higher survival rate than the bleeding group at day 90 (73.33% versus 51.85%, p=0.040). CONCLUSION: Liver cirrhosis, lower levels of fibrinogen, and major coagulation factor activity in patients with ACLF were associated with an elevated risk of bleeding events during hospitalization, which further impaired the 90-day survival rate.

High Expression of Interleukin-33/ST2 Predicts the Progression and Poor Prognosis in Chronic Hepatitis B Patients with Hepatic Flare.

BACKGROUND: Interleukin-33 (IL-33), along with its receptor suppression of tumorigenicity 2 (ST2), is capable of regulating immune responses. Immunologically mediated events play a critical role in the acute phase of chronic hepatitis B (CHB) infection. The present study primarily aimed to determine whether the IL-33/ST2 axis could be used as a reliable biomarker to predict disease progression and prognosis. METHODS: The study included 130 cases of CHB, with 48 cases in stable condition, 50 cases of progression to hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACLF), and 32 cases of progression to HBV related pre-ACLF. The demographic data and laboratory test results were recorded and compared among the groups. The blood samples for the measurement of serum IL-33 and soluble ST2 (sST2) levels were collected at admission and evaluated twice using the ELISA method. RESULTS: The patients in which the disease progressed to HBV-ACLF had the highest serum IL-33 and sST2 levels among the three groups (p<0.001). The correlation analysis showed that the serum IL-33 levels were associated with the levels of ALT (r = 0.367, p<0.001), AST (r = 0.456, p<0.001) and the MELD score (r = 0.377, p = 0.001). The area under the curve (AUC) of IL-33 and sST2 levels for differentiation of disease progression were 0.861 (95% CI: 0.787-0.934, p<0.001) and 0.788 (95% CI: 0.692-0.884, p<0.001), respectively. The serum IL-33 levels combined with the MELD score had the highest 90-day mortality prediction efficiency, with an AUC of 0.918 (95% CI: 0.859-0.977, p<0.001), a sensitivity of 92.3%, and a specificity of 88.7%. CONCLUSIONS: The IL-33/sST2 axis could be used to evaluate the progression and mortality in CHB patients with hepatic flare. The combinatorial use of multiple indicators could achieve the highest diagnostic and predictive accuracy.

Organ-associated pseudosarcomatous myofibroblastic proliferation with ossification in the lower pole of the kidney mimicking renal pelvic carcinoma: A case report.

BACKGROUND: Organ-associated pseudosarcomatous myofibroblastic proliferation (PMP) is a very rare disorder. In the urogenital tract, PMP preferentially involves the urinary bladder; kidney involvement is rare. Here, we report a rare case of PMP with ossification in the lower pole of the kidney, which mimics urothelial carcinoma or an osteogenic tumor. CASE SUMMARY: A Chinese man was admitted to our hospital due to intermittent hematuria for more than 1 mo. Enhanced renal computed tomography revealed a mass in the left renal pelvis and upper ureter. The preoperative clinical diagnosis was renal pelvic carcinoma, determined by imaging examination and biopsy. After a standard preparation for surgery, the patient underwent retroperitoneoscopic radical nephroureterectomy. The operative findings were an extensive renal tumor (6 cm × 4.5 cm × 4.5 cm) invading the lower pole of the kidney and upper ureter. The final pathological diagnosis was organ-associated PMP with ossification. After 6-mo follow-up, no recurrence or metastasis was found. CONCLUSION: This case of PMP was unusual for its mimicking renal pelvic carcinoma in imaging examinations, making biopsy necessary.

Cardiac health in patients with hepatitis B virus-related cirrhosis.

Not only alcoholic cirrhosis related to cardiac dysfunction, cirrhosis caused by nonalcoholic etiology including hepatitis B virus (HBV) infection also related to impaired cardiac health. The aims of present study were to perform a noninvasive evaluation of cardiac function and to evaluate exercise performance in HBV related cirrhotic patients without typical symptoms of cardiac disease.Seventy-nine HBV related cirrhotic patients and 103 matched subjects without a previous history of cardiac involvement were recruited. Clinical examination and cardiac health evaluation were performed. The incidence, risk factors of cardiac dysfunction and exercise tolerance were investigated.A correlation between QTc interval and model for end-stage liver disease score (R = 0.239, P = .018) was detected, however, the connection between QTc prolongation and the severity of liver disease was uncertain. Patients with HBV related cirrhosis had a tendency toward left ventricular wall thickening (P = .007). Forty-one patients (51.90%) were in accordance with the definition of cirrhotic cardiomyopathy, and a significant increase in the incidence of cardiac diastolic dysfunction (CDD) could be found with increasing Child-Pugh grade (P = .004). HBV related cirrhotic patients with CDD had a higher level of pro-brain natriuretic peptide (P = .025), international normalized ratio (P = .010) Child-Pugh score (P = .020), and a higher proportion of ascites (P < .001). The higher Child-Pugh score (odds ratio = 1.662, P = .010) was an independent diagnostic predictor of CDD. The cardiac depression and exercise tolerance also got worse with increasing Child-Pugh score (P < .001).Impaired cardiac health was common in HBV related cirrhotic patients. Cardiogenic factors must be carefully considered in the integral therapy of cirrhosis. Hepatology physicians should lay emphasis on exercise training in daily life.

A Modification of the Stapled TransAnal Rectal Resection (STARR) Procedure for Rectal Prolapse.

PURPOSE: This study was designed to assess the safety, efficacy, and postoperative outcomes of the modified Stapled TransAnal Rectal Resection (modified STARR) in patients presenting with cases of limited external rectal prolapse. METHODS: A prospective cohort of patients with mild rectal prolapse undergoing rectal resection with the Tissue-Selecting Technique Stapled TransAnal Rectal Resection Plus (TSTStarr Plus) stapler between February 2014 and September 2016 was reviewed retrospectively. RESULTS: Twenty-five eligible patients underwent rectal resection with the TSTStarr Plus stapler. The median vertical height of the resected specimen was 5.0 cm (range = 3.1-10 cm) with all cases being confirmed histologically as full-thickness resections. Over a follow-up of 33.6 ± 9.4 months, only 1 case (4%) was encountered with recurrence. The mean postoperative Wexner score was significantly improved when compared with the preoperative scores (preoperative: median = 3, range = 0-20, vs postoperative: median = 2, range = 0-20, respectively; P = .010). The median preoperative Symptom Severity Score and Obstructed Defecation Score were both decreased compared with the postoperative scores ( P = .001). CONCLUSIONS: Modified STARR in management of mild rectal prolapse appear to be a safe and effective technique. The initial results would encourage a more formal prospective assessment of this technique as part of a randomized trial for the management of mild rectal prolapse.

Risk Factors and Outcomes of Acute Kidney Injury in Patients With Hepatitis B Virus-Related Acute-on-Chronic Liver Failure.

BACKGROUND: Acute kidney injury (AKI) is common in patients with hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACLF); however, few studies concerning the risk factors and recovery patterns of renal function have been published. MATERIALS AND METHODS: A retrospective analysis of 150 patients with HBV-ACLF was performed. The occurrence, risk factors and functional recovery of AKI among patients with HBV-ACLF were investigated. RESULTS: A total of 90 patients (60%) with HBV-ACLF developed AKI. Patients with AKI had higher creatine kinase (P = 0.004), total bilirubin (P = 0.039), HBV viral load (P = 0.044), serum creatine (P < 0.001) and model for end-stage liver disease (MELD) score (P < 0.001) values and a higher proportion of hepatic encephalopathy (P = 0.032) and spontaneous bacterial peritonitis (SBP) (P = 0.042) than patients without AKI. Logistic regression analysis illustrated that SBP (odds ratio = 6.214, P = 0.012) and MELD score (odds ratio = 1.097, P = 0.006) were risk factors for the development of AKI. A subgroup analysis of recovery patterns in renal function showed that patients with a severe AKI stage had worse outcomes (P = 0.007). The proportion of patients who experienced a complete recovery was higher in survivors than in the overall AKI populations (P = 0.004). Follow-up studies showed that the no-AKI group had a higher transplant-free survival rate than the AKI group at day 90 (80.0% versus 26.7%, respectively, P < 0.001). The survival rate among patients with AKI Stage 1 was higher than that of patients with AKI Stage 2 and patients with AKI Stage 3 (P < 0.001). CONCLUSIONS: AKI is common in patients with HBV-ACLF. The SBP and MELD score have some prognosis value for patients with AKI. AKI and its stages affect the 90-day transplant-free mortality rate. It is important to focus on exploring the early recognition of AKI and early intervention of those risk factors in individuals with HBV-ACLF.

Altered intrinsic hippocmapus declarative memory network and its association with impulsivity in abstinent heroin dependent subjects.

Converging evidence suggests that addiction can be considered a disease of aberrant learning and memory with impulsive decision-making. In the past decades, numerous studies have demonstrated that drug addiction is involved in multiple memory systems such as classical conditioned drug memory, instrumental learning memory and the habitual learning memory. However, most of these studies have focused on the contributions of non-declarative memory, and declarative memory has largely been neglected in the research of addiction. Based on a recent finding that hippocampus, as a core functioning region of declarative memory, was proved biased the decision-making process based on past experiences by spreading associated reward values throughout memory. Our present study focused on the hippocampus. By utilizing seed-based network analysis on the resting-state functional MRI datasets with the seed hippocampus we tested how the intrinsic hippocampal memory network altered toward drug addiction, and examined how the functional connectivity strength within the altered hippocampal network correlated with behavioral index 'impulsivity'. Our results demonstrated that HD group showed enhanced coherence between hippocampus which represents declarative memory system and non-declarative reward-guided learning memory system, and also showed attenuated intrinsic functional link between hippocampus and top-down control system, compared to the CN group. This alteration was furthered found to have behavioral significance over the behavioral index 'impulsivity' measured with Barratt Impulsiveness Scale (BIS). These results provide insights into the mechanism of declarative memory underlying the impulsive behavior in drug addiction.

Evaluation of clustering algorithms for gene expression data using gene ontology annotations.

BACKGROUND: Clustering is a useful exploratory technique for interpreting gene expression data to reveal groups of genes sharing common functional attributes. Biologists frequently face the problem of choosing an appropriate algorithm. We aimed to provide a standalone, easily accessible and biologically oriented criterion for expression data clustering evaluation. METHODS: An external criterion utilizing annotation based similarities between genes is proposed in this work. Gene ontology information is employed as the annotation source. Comparisons among six widely used clustering algorithms over various types of gene expression data sets were carried out based on the criterion proposed. RESULTS: The rank of these algorithms given by the criterion coincides with our common knowledge. Single-linkage has significantly poorer performance, even worse than the random algorithm. Ward's method archives the best performance in most cases. CONCLUSIONS: The criterion proposed has a strong ability to distinguish among different clustering algorithms with different distance measurements. It is also demonstrated that analyzing main contributors of the criterion may offer some guidelines in finding local compact clusters. As an addition, we suggest using Ward's algorithm for gene expression data analysis.

[Prediction of ventilatory function in children and adolescents using backpropagation neural networks].

The aim of this study is to develop backpropagation neural networks (BPNN) for better prediction of ventilatory function in children and adolescents. Nine hundred and ninety-nine healthy children and adolescents (500 males and 499 females) aged 10-18 years, all of the Han Nationality, were selected from Inner Mongolia Autonomous Region, and their heights, weights, and ventilatory functions were measured respectively by means of physical examination and spirometric test. Using the approaches of BPNN and stepwise multiple regression, the prediction models and equations for forced vital capacity (FVC), forced expiratory volume in one second (FEV1), peak expiratory flow (PEF), forced expiratory flow at 25% of forced vital capacity (FEF25%), forced expiratory flow at 50% of forced vital capacity (FEF50%), maximal mid-expiratory flow (MMEF) and forced expiratory flow at 75% of forced vital capacity (FEF75%) were established. Through analyzing mean squared difference (MSD) and correlation coefficient (R) of the ventilatory function indexes, the present study compared the results of BPNN, linear regression equation based on this work (LR's equation), prediction equations based on the studies of Ip et al. (Ip's equation) and Zapletal et al. (Zapletal's equation). The results showed, regardless of sex, the BPNN prediction models appeared to have smaller MSD and higher R values, compared with those from the other prediction equations; and the LR's equation also had smaller MSD and higher R values compared with those from Ip's and Zapletal's equations. The coefficients of variance (CV) for FEF50%, MMEF and FEF75% were higher than those of the other ventilatory function parameters, and their increasing percentages of R values (ΔR, relative to R values by LR's equation) derived by BPNN were correspondingly higher than those of the other indexes. In sum, BPNN approach for ventilatory function prediction outperforms the traditional regression methods. When CV of a certain ventilatory function parameter is higher, the superiority of BPNN would be more significant compared with traditional regression methods.

Expression of human glucocorticoid receptor in T lymphocytes in acute-on-chronic hepatitis B liver failure.

BACKGROUND: Glucocorticoid receptor (GR) participates in the pathogenesis of liver inflammation. However, the potential role of GR in acute-on-chronic hepatitis B liver failure (ACHBLF) is still obscure. AIM: This present study was aimed to determine peripheral GR expression in ACHBLF patients. METHODS: Forty patients with ACHBLF, 20 patients with chronic hepatitis B (CHB) and 16 healthy controls were included in this retrospectively study. Flow cytometry was used to determine the peripheral expression of GR + T lymphocytes. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was performed for assessing relative mRNA levels of GR alpha and beta isoforms in peripheral blood mononuclear cells. Serum cortisol level was evaluated using radioimmunoassay. RESULTS: The serum cortisol level and the percentage of GR + T lymphocytes in ACHBLF patients were significantly decreased compared with CHB patients and healthy controls. However, there were no significant differences in mean fluorescence intensity (MFI) of GR + T lymphocytes within three groups. The relative GR alpha mRNA expression in ACHBLF patients was significant decreased compared with healthy controls. However, the relative GR beta mRNA expression in ACHBLF patients was significantly increased compared with CHB patients and healthy controls. In ACHBLF patients, the percentage of GR + T lymphocyte was significantly positively associated with relative GR alpha expression, prothrombin activity, and HBV DNA level, but negatively correlated with serum cortisol level and MELD score. Furthermore, the percentage of GR + T lymphocytes was also obviously elevated in survivors than non-survivors. CONCLUSIONS: It was strongly suggested that GR play an important role in the pathogenesis of ACHBLF.

[Decreased expression of serum cortisol in patients with severe hepatitis].

OBJECTIVE: To investigate the profile of serum cortisol levels in patients with severe hepatitis. METHODS: Fifty patients with viral hepatitis (including 30 severe hepatitis patients and 20 chronic hepatitis B patients) were enrolled in this present study. Serum cortisol concentration was tested using radioimmunoassay. Furthermore, liver function, coagulation and other related laboratory indices were also determined. RESULTS: Serum cortisol concentration of severe hepatitis group was lower than that of chronic hepatitis B group (P < 0.05) and lower than that of healthy controls (P < 0.05) serum cortisol concentration of severe hepatitis patients was significantly positively correlated with PTA (r = 0.445, P < 0.05); serum cortisol concentration has no relation with ALT in patients with severe hepatitis (P > 0.05), and serum cortisol concentration was significantly negatively correlated with the ratio of AST/ALT in patients with severe hepatitis (r = -0.367, P < 0.05). No significant relationship was found between serum cortisol concentration and total Bilirubin (P > 0.05). Serum cortisol concentration in death group of severe hepatitis was lower than that in survival group of severe hepatitis (P < 0.05). Of severe hepatitis patients with MELD score, the higher MELD score, the lower the cortisol concentration. CONCLUSION: Cortisol concentration decreased in patients with severe hepatitis, which was related to functional liver reserve and disease severity. Cortisol can be related to the prognosis of severe hepatitis patients.

[Standardized diagnosis and treatment of azoospermia: a report of 1027 cases].

OBJECTIVE: To constitute an appropriate procedure for clinical diagnosis and treatment for men with azoospermia in China. METHODS: Following a procedure which combined the recommendations from WHO, European Association of Urology (EAU), American Urological Association (AUA), American Society for Reproductive Medicine (ASRM) with our own experiences, 1 027 patients with azoospermia were divided into different subtypes according to patients' case histories and outcomes of various routine and specialized examinations. Then appropriate treatments were chosen according to the diagnostic subtypes. RESULTS: There were 516 cases of obstructive azoospermia, 315 of non-obstructive azoospermia and 150 of combined (obstructive and non-obstructive) azoospermia and 46 of azoospermia with special aetiology. Of the 96 patients who adopted surgical intervention, 28 underwent transurethral resection of the ejaculatory ducts, 10 vasovasostomy and 58 vasoepididymostomy. Of the 45 patients who adopted drug intervention, 16 with inflammatory obstructive azoospermia were treated with antibiotics and 29 with hypogonadotropic hypogonadism were treated with hormonal therapy. Of all the subjects, 789 patients chose in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) and 142 chose artificial insemination of donor (AID) or adoption. CONCLUSION: (1) A standard procedure should be established for the diagnosis, classification and treatment of azoospermia; (2) Special examinations and treatments should be given to cases of azoospermia with special aetiology; (3) According to the diagnosis and the patients' conditions, the most appropriate treatment options should be recommended to them; (4) The hereditary factors in azoospermia should be seriously considered, and a genetic work-up and counseling should be offered in such cases.

Stereological measurement of rat's seminiferous tubule.

BACKGROUND: The morphological measurements of seminiferous tubules are important in the studies of testis tissues. The purpose of this study was to evaluate the feasibility of using a stereological method to measure the geometric parameters of seminiferous tubule and to optimize the method. METHODS: A stereological image processing program was developed with Delphi for the stereological measurement. Fields of view were obtained from 15 healthy Wistar rats' testis tissues (n = 247). The diameter, area and volume of seminiferous tubule were estimated with the image processing program by two individual observers. The area results were compared with those obtained by the standard morphometric method of planimetry. RESULTS: Diameter measurements showed the diameters of different seminiferous tubules were almost the same and the mean value of about 50 tubules could be a good representation of the whole structure. Area measurements indicated there was no significant difference between stereology and planimetry (P > 0.05). But the stereological method required about 45% less time. Volume measurement showed the inter-observer variability was small (P > 0.05) and the reproducibility of the stereological method was good. CONCLUSION: The stereological technique was practical and efficient in the quantitative measurement of the rat's seminiferous tubule.

[Short time ECG signal analysis based on the reconstruction of phase space].

Based on the reconstruction of two-dimension phase space of time series of short ECG signals, the variation of the strange attractor geometry is described and two indices, VMI and VAI, are derived in this paper. The two indices can distinguish clearly the ECG signals of sinus rhythm, tachycardia and ventricular fibrillation. Stable results of VMI and VAI can be obtained by analyzing ECG signals of several seconds. They are expected to be used in the development of medical instruments for a fast realtime display of analysis results.

[Establishment of the database of the 3D facial models for the plastic surgery based on network].

OBJECTIVE: To collect the three-dimensional (3D) facial data of 30 facial deformity patients by the 3D scanner and establish a professional database based on Internet. It can be helpful for the clinical intervention. METHODS: The primitive point data of face topography were collected by the 3D scanner. Then the 3D point cloud was edited by reverse engineering software to reconstruct the 3D model of the face. The database system was divided into three parts, including basic information, disease information and surgery information. The programming language of the web system is Java. RESULTS: The linkages between every table of the database are credibility. The query operation and the data mining are convenient. The users can visit the database via the Internet and use the image analysis system to observe the 3D facial models interactively. CONCLUSIONS: In this paper we presented a database and a web system adapt to the plastic surgery of human face. It can be used both in clinic and in basic research.

[Development of an ECG signals analysis service system based on B/S].

An online ECG signals analysis service system based on the network has been established and is introduced in this paper which integrates the advantages of the network, database and ECG analysis tools, providing on-line ECG signal analysis service. Users with the knowledge of general operation of a browser, can process the ECG data and get a real-time display of analysis results.

Role of parasympathetic overactivity in water immersion stress-induced gastric mucosal lesion in rat.

Stress ulcer is clinically prevalent, but the underlying mechanisms are not well understood. This study aimed to investigate the role of sympathovagal imbalance in the development of water immersion restraint stress (WRS)-induced gastric mucosal lesion. Wistar rats were subjected to either restraint stress (RS) (n = 7) or WRS (n = 7) for 5 h. Linear parameters of heart rate variability and Poincaré plot were analyzed on the basis of the surface ECGs. Gastric mucosal lesion was evaluated by gross anatomy and histology. Mean R-R intervals significantly increased (P < 0.001) in a time-dependent manner in the WRS group but slightly decreased (P < 0.001) in the RS group. Root mean square of successive differences of R-R intervals and high-frequency norm (high-frequency power normalized by the total frequency power) were significantly higher in the WRS group than the RS group (P < 0.001). Low-frequency norm and low-to-high-frequency ratio increased significantly 1 h after stress and then declined to similar levels in both groups. The Poincaré plots of R-R intervals in the WRS group shifted right-upwardly and showed dispersed patterns compared with the RS group. Gastric mucosae showed serious hemorrhage, effusion, and structural collapse in the WRS group but remained normal in the RS group. Bilateral cervical vagotomy suppressed the increase of heart rate variability and prevented the gastric mucosal lesion induced by WRS. We conclude that parasympathetic overactivity is the predominant autonomic response to WRS and is most probably the leading mechanism of WRS-induced gastric mucosal lesion in rat.

Structure analysis of the protein transduction domain of human Period1 and its mutant analogs.

Human Period1 (hPer1) has been proved to be able to translocate into cells in a protein transduction manner. The segment of amino acids 830-845 of hPer1 is its protein transduction domain (PTD). In order to explore the membrane penetrating mechanism of hPer1-PTD and the physico-chemical properties necessary in the process, Ala scanning mutation method was used to investigate the variation in the peptide internalization. To further investigate the related physico-chemical requirements, the three dimensional structures of hPer1-PTD and its mutant analogs were simulated by Rosetta method. The electrostatic potentials and energies of these structures were calculated using the Delphi algorithm to solve Poisson-Boltzman equation. The hydrophobicity was assessed by the percentage of the nonpolar area in SAS (solvent accessible surface (SAS)). It has been proved that the Arg836 was the key residue for peptide internalization. When this Arg mutated into Ala, the peptide could not cross the membrane. The large enough area with positive charge was the decisive factor for hPer1-PTD. The alpha-helical structure seemed to play an assistant role so as to enable the positive charge connected in spatial arrangement.

Protein transduction domain of membrane penetrating peptide can efficiently deliver DNA and protein into mouse liver for gene therapy.

BACKGROUND: The development of a harmless and efficient nonviral gene delivery system that can facilitate the penetration of nucleic acids through the plasma membrane is a key to successful gene therapy. The aim of this study was to test a nonviral gene transferring vector's function of delivering DNA into liver cells to provide an important clue for gene transfer in liver gene therapy. METHODS: The complex of DNA and DNA delivering protein was injected into mice through their tail veins. Then the mice were killed and their liver tissue was sectioned. The gene transferring results were detected using a confocal laser scanning microscope. RESULTS: Fluorescence analysis indicated that both DNA-membrane penetrating peptide (MPP) complex and DNA- hepatocyte specific receptor binding domain (HSRBD)-MPP complex could go into liver cells. The fluorescence value of liver cells in the DNA- HSRBD-MPP group was higher than that in the DNA-MPP group. CONCLUSIONS: MPP can successfully deliver DNA and protein into cells, and MPP with a HSRBD can specifically deliver DNA into liver cells. These have laid a foundation for further study on the nonviral liver cell gene delivering system.