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Deepfake can be used to combine human voices,faces and body movements into fake content,switch gender and change age,etc.There are some problems of gender-forged face images based on generative adversarial image translation networks such as the irrelevant image domain changes easily and insufficient face details in generated images.To solve these problems,an gene-ration model of gender-forged face image based on improved CycleGAN is proposed.Firstly,the generator is optimized by using the attention mechanism and adaptive residual blocks to extract richer facial features.Then,with the aim to improve the ability of the discriminator,the loss function is modified by the idea of relative loss.Finally,a model training strategy based on age constraints is proposed to reduce the impact of age changes on the generated images.Performing experiments on the CelebA and IMDB-WIKI datasets,the experimental results show that,compared with the original CycleGAN method and the UGATIT method,theproposed method can generate more real gender-forged face images.The average content accuracy of fake male images and fake female images is 82.65% and 78.83%,and the average FID score is 32.14 and 34.50,respectively.

Based on TCM-KNN（transductive confidence machine for K-nearest neighbors） algorithm,the filter-based feature selection and cluster-based instance selection methods were used towards optimizing it as a lightweight network anomaly detection scheme,which not only reduced its complex feature space,but also acquired high quality instances for training.A series of experimental results demonstrate the two methods for optimizations are actually effective in greatly reducing the computational costs while ensuring high detection performances for TCM-KNN algorithm.Therefore,the two methods make TCM-KNN be a good scheme for a lightweight network anomaly detection in practice.

The universal re-encryption scheme URE and its extension EURE were reviewed,discovering their vulnerabilities to Pfitzmann attack.Then a novel re-encryption scheme WGRe（WonGoo Re-encryption） was proposed.WGRe could en-sure the correctness of the outputs from a node and protect against replay attack,which was hard in layered encryption sys-tems.The experimental results show WGRe is practical and more efficient than URE and EURE schemes due to the fact that its ciphertext is only double the size of the corresponding plaintext,instead of four times as URE or EURE.

The imaged could be seemed to the approximation of the polygons,so it was important to the image match that researched the polygon match method.It is based the combination of the axis of least inertia and the chain,this method was capable of preserving both contour as well as region information,and it utilized the degree of triangular membership which computed by the feature triangular to compute the similarity between the two objects.This method was invariant to image transformations,and robust to concave polygons.The experimental results show that this method’s performance and retrieval efficiency is well.

Firstly,various definitions of software assurance were discussed,the key properties of software assurance were summarized,and the software assurance model S3R（security,safety,reliability,survivability） was proposed to describe the discipline of software assurance.Then,the software assurance programs of DHS,DOD and NASA were highly in-troduced as well as S3R development.Finally,the software assurance standards-setting was specified and the develop-ment trends of software assurance research were pointed.

A quantitative method of vulnerability assessment was put forward on the foundation of research on current assessment methods home and abroad in order to evaluate vulnerability risk objectively.Vulnerability connection network（VCN） was discussed firstly for the purpose of taking connection of vulnerability into account,which divides vulnerabil-ity risk into entity risk and path risk;Secondly,advanced methods of quantifying entity importance in entity risk and network node connection effects in path risk were proposed;Lastly,an evaluation example of vulnerability risk on dis-patching management information system in power system was given out.Corresponding results show that quantitative assessment model brings out scientific and credible evaluation on vulnerability risk.

Privacy and anonymity on the Internet are gaining increasing attention from both the scientific and the large Internet user community in recent years.Privacy does not only mean the confidentiality of information,but also mean not revealing who is communicating with whom,and how often.After describing the technology of MIX,a survey of ano-nymity technologies based on it was given.Finally WonGoo,a scalable and practical decentralized peer-to-peer protocol was presented that provides strong anonymity and high efficiency with layered encryption and random forwarding.

A parametric model was established with SFE-CONCEPT software for a BIW (body in white). Firstly, the static bending-torsion stiff and the lower-order modal of BIW were simulated based on finite element method, and the results were compared with tests. Then connecting the parametric BIW with power train and chassis, the crash safety performances were simulated based on finite element method and compared with tests. The comparison results between simulation and test validate the parametric model of BIW. Secondly, the lightweight and safety of BIW structure were optimized. Due to the safety simulation analysis belongs to non-linear analysis, the optimization process can't fully achieve automatic, needing manual intervention to extract safety performances in tradition way. In order to achieve the optimization process optimization BIW can reduce 24. 17 kg, the reduction rate reaches 7. 63%. The multi-objective collaborative optimization can provide obvious lightweight effect for BIW. automation, a batch file was compiled to extract safety performances. The lightweight optimization results show that, the bending stiff can increase 0. 15%, the torsion stiff can reduce 0. 03%, torsion modal frequency can increase 1.30%, bending modal frequency increased 0.09%. And under the condition that the crash safety performances keep no changing, the weight of lightweight [ABSTRACT FROM AUTHOR]

The problem of resources and energy is increasingly getting public attention because of the development of science and technology. In this case, people have a higher demand of the portable and power- saving devices. In the post-PC era, the embedded-system devices came into being. Since the research of any problem should follow & from the whole to the part, from the simple to the complex, from the shallow to the deep" rule, embedded systems structure-the predominant topic-is worthy of exploration before handling its related isues. Despite the past research on the structure by scholars and technologists, this paper demonstrates that the social development and peoples learning process should foUow & naturalness" and & the theory of proc--s" from the perspective of philosophy. [ABSTRACT FROM AUTHOR]

Based on the transmission equation of acoustic wave in a layered media, the absorption coefficient of the multilayer porous material backed with an air gap is calculated. Taking a double-layer Aluminum foam backed with an air gap for an example, the effect of acoustic parameters of each layer and thickness of air gap on the absorption coefficient is analyzed. The results show that the absorption coefficient of the sound absorption structure with an air gap gradually increases with the increase of porosity, thickness, and flow resistivity of each porous material respectively. At low frequency, increasing the thickness of air gap will lead to the increase of the absorption coefficient, and the maximum absorption coefficient shifts towards lower frequencies. At intermediate frequency, the sound absorption structure without the air gap shows a better sound absorption property when the porosity or flow resistivity are increased. Therefore, the sound absorption level can be achieved in a wider frequency range through the adjustment of material parameters of each layer. [ABSTRACT FROM AUTHOR]

The stability of laser diode (LD) is often limited due to its temperature characteristics. The defect can lead to a decrease of output power in a parabolic rate as well as other side effects when temperature rapidly goes up. To overcome this difficulty, the paper presents a typical constant temperature control system with high-accuracy. This system performs efficiently in constant temperature controlling. The temperature signal is collected by thermistors. Then the signal processing circuit controls the semiconductor cooling chip LTC1923 at real time. Then a H-bridge is used to drive the TEC so that the LD working temperature can be controUed accurately. At the same time, for remote control of the system, the MCU is connected to PC by WiFi on which the working temperature is set up. The maximum deviation of the temperature control is only ±0.01 °C [ABSTRACT FROM AUTHOR]

Design concept and production process of chair fabric decorated in the high-grade passenger car were introduced in this paper. High quality 64 count Australian wool was blended with conductive fiber and the wool after dyeing was treated by special flame-retardant agent, so the flame retardant and anti-static function were given to this product. Double-deck and heavy design was adopted with 50 tex × 2 warp and 50 tex × 3 weft in the fabric design. The great pressure of rollers, big gauge and low speed were used in spinning, duble twist and yarn steaming. The temperature of yarn steaming was reduced 10 degrees. The product was weaven with crude steel reed and the selvage catchered by four plain weaves without the edge folding device. The pressure of crabbing and decatizing was enlarged when finishing. Many technical difficulties were solved by adopting the above technical measures and the product was produced successfully to meet the requirements. [ABSTRACT FROM AUTHOR]

In recent years, the social function of museums has been paid more and more attention by tourists and the community in China. In addition, to protect all kinds of historical artifacts, the museum also provides visitors a high-quality cultural tourism experience, which shows powerful social and educational functions of museums. Since a lot of expertise is involved in museum exhibit items, interpretation system plays an important role in visitors' understanding of the items and achieving of the social and educational functions of museums. However, planning and design of museum interpretation system in China is mainly based on expert-led perspective, largely ignored visitors' knowledge stock and demand characteristics. This causes the limited usage of the interpretation tools provided by museums to the visitors, and thus affect the experience of visitors and educational function of museums. On this basis, this paper makes the knowledge needs of visitors as a foothold and the Jinsha Musemum of Chengdu as a case, explores the visitors' expectations, usage and evaluation to the interpretation system. The results show that; (1) As Chengdu's renowned cultural museum, Jinsha site has placed a relatively high demand for knowledge from public visitors. Its interpretation system also has placed high expectations, not only for richness of knowledge, but also on knowledge transfer process and methods. (2) Visitors' pre-visit expectations for knowledge needs and interpretation system have a significant positive correlation with post-visit knowledge gain and evaluation of interpretation system. Visitors with higher expectation will be more active to interpretation system and eventually get a better experience of knowledge. However, there is a gap between actual perception and expectations of visitors, expectation is only partially achieved. This gap is negatively correlated to the expectation and positively correlated to the actual perception. This indicates that the quality of interpretation system in the Jinsha Museum exceeds the expectations of visitors with lower expectations but not yet reached the expectations of visitors with higher expectations. (3) Price, habit, and promoting or inhibiting effect among all kinds of interpretation tools mainly affect the difference of use proportions for tools. (4) The use of the narrator can effectively improve the satisfaction of the visitors to artistic appeal and popularity of the interpretation. Which shows that on-site exchange between people and face to face counseling is still an important means of acquiring knowledge. In addition, the contents of the explanatory plaques, panels and touch screen are too professional to fully understand by visitors who don't have specialized knowledge. This limited knowledge gain of the visitors and thus affecting their evaluation of interpretation system. (5) The satisfaction with the interpretation tools significantly affect knowledge gain of visitors and their evaluation of the interpretation system. At the same time, the higher the satisfaction of interpretation system, the smaller the gap between the evaluation and the expectation, that means the improvement of tools quality is an effective way to satisfy visitors' expectation and reduce the gap between the evaluation and the expectation. There are some limitations of this paper; (1) Only one case was investigated which may influence the reliability and validity of the study results. The study procedure should be duplicated in other museum to form a more thorough understanding of the research topic. (2) Visitors' perception has a strong subjectivity and sometimes may not be consistent with the actual knowledge gain. How to measure the objective knowledge gain of visitors, and how the relationship between objective knowledge gains and subjective perception, need to be studied in future research. (3) This study obtained some valuable results on the Jinsha Museum. How to use these results to improve the interpretation system needs to be further explored. [ABSTRACT FROM AUTHOR]

Aiming at the disadvantages of traditional AMD drive device, such as large volume, high cost, complex system, more energy consumption, an innovative driver-Direct Driving Volume Control (DDVC) servo system for AMD is proposed based on the volume control of electro-hydraulic servo system. The mechanical models of motor control, hydraulic power mechanism, hydraulic actuator and other sub-structures of DDVC e-lectrohydraulic servo system were separately established as well as the mathematical model of direct drive volume control electro-hydraulic servo actuator. Taking an actual project's experimental prototype as a standard, the parameter optimization and the match computation were carried out, and the DAMD control system are designed and processed. The dynamic characteristics of the power mechanism controlled by pump are not high because the DAMD system dynamic characteristics are low. DAMD control system can be used to substitute the traditional AMD control system for high-rise structure or large-span structure to realize active control of structural vibration. [ABSTRACT FROM AUTHOR]

A leg-wheeled passive wheel type of biped ice-skater robot was designed on the basis of rolling and skating principle in this paper. The kinematic analysis on it was carried out by Denavit-Hartenberg (D-H) method. The robots legs were classified into supporting leg and swing leg. Assumed that the static frictional force in the normal direction of four wheels on the swing leg was in critical condition, three kinds of stability criteria taking the kinematic constraints of the wheel and the balance of the body into account were discussed depending on the state between the wheels and the plane. The validity and feasibility of the stability criterion were verified by simulations and experiments on the robot. [ABSTRACT FROM AUTHOR]

The metallic foam core of lightweight composite sandwich plates undergoes significant deformation along the thickness direction under shock loading, and hence the transverse normal and shear compressions of the core must be considered. A higher-order theory is used to analyze the transverse normal and shear strains of the core, whilst the Kirchhoff theory is used to analyze the thin composite facesheets because of their high-stiffness. The Hamilton's principle is used to obtain the equations governing the vibration performance of the sandwich plate. The extended Galerkin's method is used to solve the governing equations to obtain the vibration equation of the sandwich plate suitable for numerical analysis. The fourth-order Runge-Kutta method is employed to solve the transverse dynamic displacement of the sandwich plate within the elastic range. The inherent frequency of the sandwich plate is compared with the results of finite element method. Finally, the capacity to absorb energy of the sandwich plate is discussed by using the higher-order theory, and the influence of key system parameters is explored. The results show that the whole structural stiffness is affected by changing the ply angle of the facesheets and the thickness of the core, influencing the vibration characteristic of the sandwich composite; the structural vibration is weakened as energy is dissipated by the material damp; and the metal foam core absorbs most of the energy due to the transverse normal and shear compressions. [ABSTRACT FROM AUTHOR]

Objective: To explore the correlation between peripheral blood B cell count and clinical features and prognosis of patients with newly diagnosed diffuse large B-cell lymphoma (DLBCL)., Methods: The relationship of peripheral blood B cell count with clinical features, laboratory indexes and prognosis in 67 patients with newly diagnosed DLBCL was retrospectively analyzed., Results: Patients were divided into low B-cell count group (B cell＜0.1×10 9 /L, n =34) and high B-cell count group (B cell≥0.1×10 9 /L, n =33) according to the median B cell count values. Compared with the high B cell count group, the low B cell count group had a higher proportion of patients with Lugano stage III-IV, elevated LDH, elevated β 2 -MG and IPI score 3-5 and increased CRP ( P =0.033, 0.000, 0.023, 0.001, 0.033). The peripheral CD3 + and CD4 + cell counts of patients in the low B cell count group were significantly lower than those in the high B cell count group ( P =0.010, 0.017). After initial treatment, overall response rate (ORR) and complete remission (CR) rate in high B cell count group were significantly higher than those in low B cell count group ( P =0.032, 0.013). The median follow-up time of patients was 23(2-77) months, progression-free survival (PFS) and overall survival (OS) of patients in the high B cell count group were significantly better than those in the low B cell count group ( P =0.001, 0.002). Univariate analysis showed that pretreatment low B cell count in the peripheral blood was associated with shortened PFS and OS ( HR =4.108, P =0.002; HR =8.218, P =0.006). Multivariate analysis showed that low B cell count was an independent prognostic factor for shortened PFS ( HR =3.116, P =0.037)., Conclusion: Decreased peripheral blood B cell count in newly diagnosed DLBCL patients is associated with high-risk clinical features and may affect the efficacy of immunochemotherapy, which is associated with poor clinical prognosis.

This study explored the mechanism of the ultrafiltration extract of Angelicae Sinensis Radix and Hedysari Radix in ameliorating renal fibrosis in the rat model of diabetic kidney disease(DKD) based on the expression of hypoxia-inducible factor-1α(HIF-1α)/vascular endothelial growth factor(VEGF) and HIF-1α/platelet-derived growth factor(PDGF)/platelet-derived growth factor receptor(PDGFR) signaling pathways in the DKD rats. After 1 week of adaptive feeding, 50 male SPF-grade Wistar rats were randomized into a blank group(n=7) and a modeling group. After 24 h of fasting, the rats in the modeling group were subjected to intraperitoneal injection of streptozocin and fed with a high-sugar and high-fat diet to establish a DKD model. After modeling, the rats were randomly assigned into model(n=7), low-dose ultrafiltration extract(n=7), medium-dose ultrafiltration extract(n=7), irbesartan(n=8), and high-dose ultrafiltration extract(n=8) groups. After intervention by corresponding drugs for 12 weeks, the general conditions of the rats were observed. The body weights and blood glucose levels of the rats were measured weekly, and the 24 h urinary protein(24hUP) was measured at the 6th and 12th weeks of drug administration. After the last drug administration, the renal function indicators were determined. Masson staining was employed to observe the pathological changes of the renal tissue. The expression of prolyl hydroxylase domain 2(PHD2) and HIF-1α in the renal tissue was detected by immunohistochemistry(IHC). Real-time qPCR was employed to determine the mRNA levels of PHD2, VEGF, PDGF, and PDGFR in the renal tissue. Western blot was employed to determine the protein levels of HIF-1α, VEGF, PDGF, and PDGFR in the renal tissue. The results showed that compared with the model group, drug administration lowered the levels of glycosylated serum protein(GSP), aerum creatinine(Scr), and blood urea nitrogen(BUN) in a dose-dependent manner(P<0.05 or P<0.01) and mitigated the pathological changes in the renal tissue. Furthermore, drug administration up-regulated mRNA level of PHD2(P<0.05 or P<0.01), down-regulated the mRNA levels of VEGF, PDGF, and PDGFR(P<0.05 or P<0.01) and the protein levels of HIF-1α, VEGF, PDGF, and PDGFR(P<0.01) in the renal tissue, and increased the rate of PHD2-positive cells(P<0.01). In conclusion, the ultrafiltration extract of Angelicae Sinensis Radix and Hedysari Radix effectively alleviated the renal fibrosis in DKD rats by inhibiting the expression of key proteins in the HIF-1α signaling pathway mediated by renal hypoxia and reducing extracellular matrix(ECM) deposition.

Objective: To develop a novel objective standardized endoscopic skill training and assessment system based on artificial intelligence technology., Methods: By designing five basic skill parts of endoscopic operation including vision location, clamping, delivering, shearing and suturing, we achieved objective standardized indexes which gained automatically with image recognition and refined perception., Results: With Huaxi intelligent endoscopic skill training system, the accurate rates of vision location, clamping, delivering, shearing and suturing were 90%, 95%, 99%, 90%, and 89%, respectively. The response and performance time were 8-10 s, <1 s, <1 s, 1-3 s, and <1 s, respectively., Conclusion: Huaxi intelligent endoscopic skill training and assessment system has preliminarily possessed the capability to assess the endoscopic skills of surgeons objectively., (Copyright© by Editorial Board of Journal of Sichuan University (Medical Science Edition).)

Objective: To preliminary study of the resistance mechanisms of S. pneumoniae (S. pn) by determining the resistance rates and gene of S. pn isolated from the lower respiratory tract infection infants., Methods: Drug susceptibility test with disk diffusion and broth micro-dilution was conducted to evaluate the resistance rates of 73 strains of S. pn isolated from the lower respiratory tract infection infants to penicillin, levofloxacin and other 10 antibiotics. PCR method was used to analysis the antimicrobial resistant genes tet M, mef A, erm A, erm B and int Tn of the isolates., Results: The antibiotic resistance rates of the S. pn isolates to erythromycin, clindamycin and tetracycline were 95. 9%, 94. 5%, 87. 7% and 0% to vancomycin when tested with disk diffusion method. The antibiotic resistance rates of these isolates to penicillin, cefotaxime and ceftriaxone were 45. 2%, 47. 9% and 46. 6% respectively when tested with broth micro-dilution method. The carrier frequencies of tet M, mef A, erm A, erm B, int Tn genes in the 73 isolates were 91. 8%, 63. 0%, 58. 9%, 39. 7% and 61. 6% respectively., Conclusion: The S. pn strains isolated from infant respiratory tract in Chengdu perform a serious drug resistance problem, especially to routine antibiotics like erythromycin, clindamycin and tetracycline and cephalosporin, the resistance rate to levofloxacin, chloramphenicol remained at a low level; the resistance to tetracycline was closely related with the tet M gene fragment, the resistance to macrolide was mainly decided by active efflux pump and secondarily by the alternation of gene targeting, int Tn had close relation with tet M, erm B.

Objective: To understand the variation of G glycoprotein gene of human respiratory syncytial virus (HRSV) obtained from Sichuan in 2010 and determine the dominant genotypes., Methods: G glycoprotein gene of seven cases of subtype A and eleven cases of subtype B of HRSV were amplified by RT-PCR and sequenced. The phylogenetic trees were constructed to determine the subtype of samples. And then, the genetic variations of the second hypervariable region of G glycoprotein gene were studied., Results: The nucleotide genetic distances of G glycoprotein gene in subtype A and subtype B HRSV were 0.022 +/- 0.005 and 0.073 +/- 0.010, respectively. Transitions were more prevalent than transversions, GA -AG were the most frequent transitions detected among group A viruses, while UC+CU transitions were the most among group B. Phylogenetic analyses demonstrated that 7 subtype A virus could be clustered into one genotype, genotype GA2, and 11 subtype B virus could be clustered into two genotypes, GB2 and BA. The length of G protein gene in group A was all 298aa, but in group B included 295aa, 312aa and 315aa. Selective pressure was purifying selection in both subtypes. 9 positively selected sites in group A and 1 in group B on the second hypervariable region of G protein were identified., Conclusion: GA2, GB2 and BA were the main genotype. The changes may favor virus escape from the host immune response including the variation of the G protein gene length, frequency of nucleotide changes and selective pressure.

Objective: To investigate the effects of Jiangu granule containing serum (JGG-serum) on the cyclins in rat's osteoblast at G1 phase., Methods: Osteoblasts isolated by enzymatic digestion from SD rats were cultured and intervened with JGG-serum or normal saline (as control) respectively. Cell generation cycle was detected by flow cytometry, and expressions of Cyclin D1, cyclin-dependent kinase 4 (CDK4), oncogene protein (P21) in the osteoblast were detected dynamically using immuno-cytochemical and RT-PCR technique., Results: As compared with the control, the cell generation cycle and cell proliferation were proceeding quicker in the JGG-serum (20%) intervention group; with higher protein and mRNA expressions of Cyclin D1 and CDK4, as well as much lowered expressions of P21 in nuclei of osteoblast detected at all time points (24 h, 48 h and 72 h after treatment, P < 0.05 or P < 0.01)., Conclusion: JGG-serum can adjust the G1 phase cyclins in osteoblast cultured in vitro, increase the mRNA and protein expressions of Cyclin D1 and CDK4, and inhibit P21 expression, so as to accelerate the proliferation of osteoblast.

Objective: To investigate natural spontaneous menopausal age, menstruation span and their relationship with menarche age and parity in Pudong district of Shanghai., Methods: From Jan 2007 to Jul 2008, 15 083 spontaneous menopause women undergoing cervical cancer screening were enrolled in this study. The questionnaire included menarche age, parity, spontaneous menopausal age and menstruation span. Those women were divided into four groups based on age, which were group of 56 - 60, 61 - 65, 66 - 70 and more than 70.Analysis of variance (ANOVA) was used for comparing difference between menopausal age and menstruation span. Multiple factor regressions was used to analyze the relationship between menarche age, parity and menopausal age and menstruation span., Results: (1) Spontaneous menopausal age: the minimum was 29 years old, the maximum was 61 years old, and the mean age was (50.6 ± 3.7) years old. The mean spontaneous menopause age were (50.9 ± 3.4), (50.7 ± 3.7), (50.0 ± 4.1), (49.6 ± 4.0) years in groups of 56 - 60, 61 - 65, 66 - 70 and more than 70 years. With the increasing age range in four groups, the increasing trends of menopausal age were observed, which the difference of 1.36 year was shown between groups of 56 - 60 and more than 70 years. (2) Menstruation span: the mean of menstruation span was (34.3 ± 4.1) years, which the minimal age of 12 years and maximal age of 48 years were recorded. (34.6 ± 3.8), (34.3 ± 4.1), (33.9 ± 4.6), (33.2 ± 4.5) were observed in groups of 56 - 60, 61 - 65, 66 - 70 and more than 70 years. With the increasing age range in four groups, the increasing trends of menstruation span were observed, which the difference of 1.41 year was shown between groups of 56 - 60 and more than 70 years. (3) The impact of menarche age on menopausal age and menstruation span: there was no correlation between menarche age and menopausal age (r = 0.02); however, menstruation span was found to be negatively correlated with the menarche age (r = -0.43). (4) The impact of parity on menopausal age and menstruation span: the mean menopausal age of women who had 1 - 2 deliveries was significantly higher than those had no delivery or more than 3 deliveries (P < 0.05). However, there was no difference in menopausal age between women with 1 and 2 deliveries or between women without delivery and more than 3 deliveries (P > 0.05). Menstruation span of women with 1 delivery was significantly longer that those with more than 1 delivery (P < 0.05), similarly, women with 2 deliveries had longer menstruation span than women without delivery or more than 3 deliveries (P < 0.05). There were no difference in menstruation span between women with more than 3 deliveries and without delivery (P > 0.05). (5) Multifactor regression analysis for menstruation span: menarche age was correlated with menstruation span negatively (r = -0.97, P < 0.001). There was significantly different menstruation span between group of 61 - 65, 66 - 70 or more than 70 years and group of 56 - 60 (r = -0.18, P = 0.020; r = -0.78, P < 0.001 and r = -1.23, P < 0.001). Menstruation span in women with 1 - 2 deliveries was significantly longer than that of women without delivery or more than 3 deliveries. (6) Multifactor logistic analysis of menopausal age: there was no association between menarche age and menopausal age, however, significant differences were found in mean menopausal age between different groups, which show that menopausal age of group 56 - 60 years was significant higher than the other groups, including age-group 61 - 65 years, 66 - 70 years and over 70 years (r = -0.18, P = 0.020; r = -0.78, P < 0.001; r = -1.23, P < 0.001). Menopausal age in women with 1 - 2 deliveries was significantly higher than those of women without delivery or with more than 3 deliveries, however, no difference between women with 1 and 2 deliveries or between women without deliveries and more than 3 deliveries was observed., Conclusion: (1) Menopausal age and menstruation span exhibited increasing trends in Pudong district of Shanghai. (2) Menarche age and parity were the important factors influencing menopausal age and menstruation span. (3) With younger age of menarche, the menstruation span become longer.(4) Deliveries of 1 - 2 times can significantly delay the menopause and prolong menstruation span, however, the multiple deliveries (≥ 3 times) had no significant impact on menopausal age and menstruation span.

Objective: To survey age of menarche in Pudong district in Shanghai., Methods: Data in this study were derived from 56,924 women at age of 20-81 years in screening for cervical cancer between January 2007 and July 2008 in Pudong district. The age of menarche were recorded in a questionnaire. To investigate the trends in age at menarche in different socioeconomic status, the subjects were divided into 12 groups in 5-year birth cohorts. The mean menarche age in each group was analyzed by analysis of variance (ANOVA). The percentage of menarche age at 10-12 years and more than 18 years was analyzed by chi2 method., Results: (1) The minimum age of menarche recorded is 10 years old, and the maximum is 28 years old, with average age of menarche at 15.7 years. In all groups, the smallest average age of menarche is 14. 6 years in 26-30 years old age group, while the biggest average age of menarche age is 16.5 years in >75 years old group; The difference showed statistical significance (P < 0.01). (2) The percentages for participants with early menarche age (10-12 years old) or late menarche age (>18-year-old menarche) were 1.82% (1034/56,924) and 5.20% (2959/56,924) respectively. However, the maximum percentage for early menarche was recorded in 31-35 years old group (4.45%, 197/4431), only 0.84% (10/1191) of participants in >75 years old group was classified as early menarche. Meanwhile, the lowest percentage for late menarche was 0.38% (17/4431) in 31-35 years old group, and the highest percentage was 14.70% (91/619) in >75 years old group. The changes in the percentages for early menarche or late menarche are significantly associated with age differences (P < 0.01)., Conclusion: The study suggested that the average of onset age of menarche in Pudong district has declined over the past decades in an age-based way, accompanied with the increase of the percentage for early menarche and the decrease of percentage for late menarche.

Objective: To analyze the effect of interleukin 2 (IL-2) on immunoglobulin G(IgG) expression in cervical cancer cell line HeLaS3., Methods: By immunofluorescence-flow cytometry(FCM), we detected membrane and cytoplasmic IgG expression variation of HeLaS3 after IL-2 stimulation. By Western blot, we detected IgG expression variation of HeLaS3 after IL-2 stimulation. Meanwhile, by RT-PCR with primers designed to amplify four subclasses of IgG, we detected Gamma transcript variation in HeLaS3 after IL-2 stimulation at mRNA level., Results: The result of immunofluorescence-FACS showed that after IL-2 stimulation for 48 hours, membrane and cytoplasmic IgG expression elevated in HeLaS3. The result of Western blot showed after IL-2 stimulation for 48 hours, IgG expression elevated in HeLaS3. Meanwhile, the result of RT-PCR showed that Gamma1 transcript mildly elevated after IL-2 stimulation for 48 hours., Conclusion: IL-2 can promote IgG expression in cervical cancer cell line HeLaS3.

Objective: To investigate the association of single nucleotide polymorphisms (SNPs) of the peptidylarginine deiminase IV (PADI4) and HLA-DRB1 shared epitope (SE) alleles with rheumatoid arthritis(RA) in a Chinese population., Methods: Four exonic SNPs of the PADI4 gene (PADI 4&#95;89*A/G, PADI 4&#95;90*C/T, PADI 4&#95;92*C/G and PADI 4&#95;104*C/T) were genotyped in 67 unrelated patients with RA and 81 healthy controls, using cDNA sequencing and T vector cloning. HLA-DRB 1*01, *04 and *10 subtypes were determined by polymerase chain reaction with sequence specific primers (PCR-SSP)., Results: The distributions of the 4 SNPs were different in the two groups, and increased RA susceptibility was significantly associated with the minor alleles of PADI 4&#95;89*G (P was 0.023), PADI 4&#95;90*T (P was 0.004), PADI 4&#95;104*T (P was 0.003), and the haplotypes carrying the 4 minor alleles (P was 0.008). HLA-DRB1 SE alleles are composed of HLA-DRB 1*0101, *0102, *0401, *0404, *0405, *0408, *0409, *0410 and *1001. Individuals carrying the SE alleles were associated with increased RA susceptibility (P was 0.002). Individuals carrying both the SE alleles and minor alleles of the 4 SNPs were more susceptible to RA than individuals carrying neither the minor SNP alleles nor the SE alleles., Conclusion: The PADI4 SNPs and haplotypes are associated with RA susceptibility in Chinese. HLA-DRB1 shared epitope is also an important risky factor for RA. There may exist certain synergistic effect between the PADI4 minor alleles and the HLA-DRB1 shared epitope.

Objective: To study the correlations between the bone quality and level of u-DPD and serum 1,25-dihydroxyvitamin D3 in the ovariectomized osteoporosis rats., Methods: Forty female rats were randomly divided into the ovariectomy group (the OVX group) and the sham-operation group (the Sham group), and draw materials in 12 and 24 weeks after operation. Bone mineral density (BMD) of the 1st lumbar, the femoral neck and femur was measured by dual-energy X-ray absorptionetry. The level of urine DPD was measured by ELISA. The concentration of 1,25-dihydroxyvitamin D3 in serum was measured by radioimmunoassay (RIA). The limit of compressive strength and elastic modulus of 1st lumbar and bending load of right femur were measured by photoelasticity meter. The osseous morphology of upper tibia was measured by the non-decalcifing section. The correlation of experimental data was analyzed by statistical method., Results: In ovariectomy group, except the level of urine DPD/Cr was increase, all data decreased significantly compared with the samp group,and descending continually with the lapse of time. The results showed that there was a negative correlation between the urine DPD/Cr and BMD of the 1st lumbar, femur, the limit of compressive strength of 1st lumbar, bending load of femur and trabecular bone area. But there was a positive correlation significantly between the concentration of 1,25-dihydroxyvitamin D3 in serum and BMD of the 1st lumbar, the femoral neck and femur, trabecular bone area, the limit of compressive strength of 1st lumbar, bend load of femur., Conclusion: The correlation is remarkable between the bone quality and the level of urine DPD/Cr, 1,25-dihydroxyvitamin D3 in serum. We can predict the bone quality by measuring the level of urine DPD/Cr and 1,25-dihydroxyvitamin D3 in serum.

Objective: To investigate the suppression effect and apoptosis of the colon cell line HT-29 treated with adenovirus-expressed human hyperplasia suppressor gene (hHSG) and hyperthermia., Methods: Replication-defective recombinant adenovirus containing hHSG was constructed, and then infected into HT-29 cell line to detect its overexpression by western blot. The synergic effect with hyperthermia was observed, cell survival was assessed using Vi-CELL TMXR Cell Viability Analyzer, and cell apoptosis and cell cycle distribution were determined by flow cytometry., Results: After the cells were treated with 100 MOI (multiplicity of infection) Ad5-hHSG and heated at 42 degree C for 1 h, the number of viable cells decreased sharply, compared with single treatment with Ad5-hHSG or 42 degree C for 1 h (P < 0.01). The apoptotic rate of combined groups was significantly higher than that of Ad5-hHSG group or the heated group respectively (P < 0.01). Moreover, the combination treatment of Ad5-hHSG and 42 degree C for 1 h induced G2/M arrest., Conclusion: Ad5-hHSG could increase the sensitivity of HT-29 to hyperthermia.

Aim: To investigate the allergen composition in the species of shrimp and crab., Methods: The main allergens of shrimps and crabs were analyzed by SDS-PAGE and Western blot assay., Results: The molecule weight of main allergic protein in shrimps was near to 36 000, and the main allergic proteins in different kinds of crabs were about 29 000, 36 000, 66 000, 89 000 molecule weight., Conclusion: There are common allergens among different kinds of shrimps and crabs.

Aim: To explore the prokaryotic expression of the extracellular region of human CD1d (hCD1d) and prepare its polyclonal antibody., Methods: The gene encoding the extracellular region of hCD1d was amplified by PCR and cloned into prokaryotic expression vector pET28, then expressed in E.coli BL21 (DE3) with IPTG induction. The recombinant protein was purified by Ni2+-NTA agarose column and then used as immunogen to immunize the mouse. The generated polyclonal antibody was evaluated by ELISA, Western blot and immunohistochemical staining (IHC), respectively., Results: The recombinant extracellular region of hCD1d was successfully expressed and purified. The polyclonal antibody with high titer and high specificity was obtained, which could recognize the native hCD1d in the human small intestinal tissues., Conclusion: The recombinant extracellular region of hCD1d has been obtained. The antibody with high titer and high specificity against the extracellular region of hCD1d from the mouse has been successfully prepared, which lays a foundation for further research into the detection and functional study of CD1d.

Aim: To prepare the rabbit antibody against the alpha3 domain of the human CD1d (hCD1d-alpha3)., Methods: The gene fragment coding for hCD1d-alpha3 was amplified by PCR and cloned into prokaryotic expression vector pET28, then expressed in E.coli BL21(DE3). The recombinant protein hCD1d-alpha3 was purified with Ni(2+)-NTA agarose column and used as immunogen to immunize the rabbit. The titer and specificity of the anti-hCD1d-alpha3 antibody from the rabbit were analyzed by ELISA, Western blot and immunohistochemistry, respectively., Results: The recombinant hCD1d-alpha3 was successfully expressed and purified, and the polyclonal anit-hCD1d-alpha3 antibody was successfully prepared. The titer of the antiserum was 1:6 400 by ELISA. Western blot analysis showed this antiboday reacted specifically with hCD1d. Immunohistochemistry analysis showed the antibody could recognize the native hCD1d in the human intestinal tissue., Conclusion: The anti-CD1d-alpha3 antibody from the rabbit with high titer and specificity has been prepared with purified recombinant hCD1d-alpha3 as immunogen, which lays a foundation for further research into detection and functional study of CD1d.

Aim: To express the human mast cell chymase cDNA in E.coli and prepare the antibody against human mast cell chymase with recombinant chymase., Methods: The human mast cell chymase cDNA was cloned by RT-PCR. The recombinant chymase was expressed in E.coli with L-Arabinose induction and purified by Ni-NTA agarose column. Then the purified chymase was used as immunogen to immunize the rabbit. The titer and specificity of the anti-chymase antibody from the rabbit were analyzed by indirect ELISA and Western blot, respectively., Results: The recombinant chymase was successfully expressed in E.coli, and the polyclonal anit-chymase antibody was prepared by immunizing the rabbit with the purified recombinant chymase. The titer of the generated antiserum was detected to be 1:12 800 by ELISA. Western blot analysis showed this antibody bound specifically with chymase., Conclusion: The anti-chymase antibody from the rabbit with high titer and specificity has been prepared with purified recombinant chymase as immunogen, which lays a foundation for further research into detection and function of chymase.

Aim: To prepare antibody against human mast cell carboxypeptidase (hMC-CP) by using recombinant hMC-CP expressed in E.coli, and to characterize the antibody., Methods: hMC-CP was expressed in E.coli with L-Arabinose induction and purified through Ni-NTA column. The purified hMC-CP as immunogen was used to immunize rabbit. The titer and the specificity of the rabbit anti-hMC-CP antibody was analyzed by indirect ELISA and Western blot respectively., Results: The hMC-CP was successfully expressed and purified. The polyconal anit-hMC-CP antibody was prepared by immunizing rabbit using the purified recombinant protein, The titer of the generated antiserum was 1:6 400 by ELISA. Western blot analysis showed that this antibody could bind with hMC-CP specifically., Conclusion: The rabbit anti-hMC-CP antibody with high titer and high specificity has been prepared by using purified recombinant hMC-CP as immunogen, which lays the foundation for further research on detection and function of hMC-CP.

The effects of hypocrellin A (HA) on the conformational changes of hemoglobin and myoglobin were studied using synchronous fluorescence spectroscopy. The results indicated that HA can change the conformation of these two proteins, leading to the change in the micro-environment of tryptophane and tyrosine residues from hydrophobic environment to hydrophilic environment to different extent.

Terbium-lanthanum (or gadolinium)-poly(N-vinylacetamide) complexes were synthesized and characterized by UV-Vis absorption spectroscopy, FTIR, XPS and fluorescence spectroscopy. The results of UV-Vis, FTIR and XPS suggested that terbium and lanthanum (or gadolinium) ions were bonded to amide group of PNVA polymer. Fluorescence experiment indicated that the characteristic emission intensity of terbium ion was greatly increased and possibly sensitized by lanthanum ion (or gadolinium). Moreover, the wavelengths of terbium characteristic emissions were changed slightly. Also the characteristic emission intensity of terbium ion doped with lanthanum ion was better than that with gadolinium ion.

Background & Objective: The disturbance of cell differentiation is an important characteristic of tumor, but, up to date, the researches on this aspect are not as deep as those on cell proliferation of tumor. This study was to detect the alterations of cell differentiation and relative regulating factors during the carcinogenesis of cervical epithelia., Methods: The protein expression of keratin (KT) 10, 14, 19 and beta1-integrin and beta-catenin in 20 specimens of normal squamous epithelium (NSE), 19 specimens of dysplasia squamous epithelial hyperplasia (DSEH), 39 specimens of squamous carcinoma in situ (SCIS), and 20 specimens of invasive squamous epithelial carcinoma (ISEC) were detected by inmmunohistochemistry; the mRNA expression of of beta1-integrin and beta-catenin were detected by in situ hybridization., Results: The strong positive rates of KT10 in NSE, DSEH, SCIS, and ISEC were 85.0%, 52.6%, 18.0% and 0, respectively, with a descending trend; it was significantly lower in SCIS and ISEC than in NSE and DSEH (Chi(2)=11.28, P<0.05; Chi(2)=8.53, P<0.05). The expression of KT14 and KT19 showed a heterogeneity in SCIS and ISEC: the proteins were negative in some cases, but overexpressed in other cases with positive cells moved upwards from the basal layer; positive cells scattered at the full epithelial layer of SCIS. The expression of beta1-integrin also showed a descending trend in NSE, DSEH, SCIS, and ISEC. The positive rate of beta1-integrin was significantly lower in ISEC than in NSE and DSEH (Chi(2)=7.62, P <0.05; value of exact probability=0.014, P < 0.05); the mRNA expression of beta1-integrin showed the same trend as its protein expression in the samples although the difference were not significant. The protein and mRNA expression of beta-catenin was located in nuclei or cytoplasm of SCIS and ISEC; increased positive cells moved upwards from the basal layer., Conclusions: Restraint of terminal differentiation of epithelial cells may relate to the abnormal expression of beta1-integrin and the dysfunction of Wnt signaling pathway on regulating cell differentiation during the carcinogenesis of cervical epithelia. The expression of KT10 and beta1-integrin in SCIS is not obviously different from that in ISEC.

The interaction of Mn2+ and poly (N-isopropylacrylamide) (PNIPAAm) was studied by using UV-Vis, FTIR and fluorescence spectroscopic methods. The results indicated that Mn2+ could be bonded to oxygen atoms of carbonyl in PNIPAAm and form the complex of Mn2+ -PNIPAAm. It was found that there existed efficient Förster energy transfer from Mn2+ to PNIPAAm due to the emission spectra of Mn2+ overlapping the excitation spectra of PNIPAAm and that the emission peak of Mn2+ at 561 nm disappeared in Mn2+ -PNIPAAm complex. Therefore the fluorescence intensity at 307 nm was increased by 314%.

The interaction mechanism of thionine (TH) and calf thymus DNA (CT-DNA) was studied with UV-Vis absorption spectoscopy, fluorescence spectroscopy, circular dichroism (CD), and X-ray photoelectron spectroscopy (XPS). It was demonstrated that in the pH 7.2 phosphate buffer solution, the interaction of TH and CT-DNA is in the intercalative mode. The absorbance of the absorption peak of TH decreases and the peak position shifts to the red direction. The binding constant (K) of TH with CT-DNA is 1.45x 10(4) mol x L(-1). The fluorescence intensities of TH decrease drastically with increasing the concentration of CT-DNA. The Stem-Volmer quenching constant (Ksv) was calculated to be 1.01 x 10(4) mol x L(-1). The intercalative sites are mainly located at the G-C sequences of the CT-DNA molecule through the S atom in the TH molecule according to the experimental data obtained from the fluorescence spectra and XPS analysis. The secondary structure conformation of CT-DNA is changed due to its interaction with TH.

Oxygen of two propionic acid groups of the heme group in an MP-8 molecule, leading to an increase in the non-planarity of the porphyrin ring of the heme group. The increasing extent in the non-planarity of the porphyrin ring of the heme group due to the interaction of La3+ and MP-8 molecule would be significantly decreased because NaCl could increase the aggregation extent of MP-8 molecules. Although methanol can reduce the aggregation extent of MP-8 molecules, it has little influence on the increasing extent in the non-planarity of the porphyrin ring of the heme group due to the interaction of La3+ and MP-8 molecule.

The interaction between CT-DNA immobilized on the positively charged nylon membrane and cytochrome C was studied using UV-Vis reflectance absorption spectroscopy and electrochemistry. It was found that cytchrome C can be adsorbed on the negative charged CT-DNA surface with the active positive charge area. Therefore, CT-DNA can promote the direct electrochemical reaction of cytochrome C.

Background & Objective: Gap junction intercellular communication (GJIC), mediated by connexin (CX), is the unique type of intercellular communication in carcinoma in situ (CIS). Changes in expression of CXs and function of GJIC may play roles in carcinogenesis of cervical cancer and progression of CIS. This study was to investigate the expression of CXs in normal epithelium (NE), hyperplasia, CIS, and invasive carcinoma (IC) of cervix, to explore correlation of expression of CXs to pathogenesis and progression of cervical CIS., Methods: Expression of CX43 (mRNA, protein), CX26 (mRNA), and Ki-67 (protein) in 30 specimens of cervical NE, 22 specimens of cervical hyperplasia, 30 specimens of cervical CIS, and 26 specimens of cervical IC were detected by immunohistochemistry and in situ hybridization., Results: In cervical NE, hyperplasia, CIS, and IC, positive rates of CX43 protein were 83%, 55%, 50%, and 30%, respectively, and positive rates of CX43 mRNA were 97%, 64%, 58%, and 46%, respectively; positive rates of CX26 mRNA was 93%, 68%, 55%, and 42%, respectively. Positive rates of CX43 (mRNA, protein) and CX26 (mRNA) were gradually decreased. Positive rate of CX was significantly lower in CIS group than in NE group (P < 0.05). The expressions of CX26 and CX43 were weaker in CIS group than in NE groupu their expressions were obviously decreased, or even vanished in the area of CIS adjacent to NE. Proliferating index (PI), indicated by expression of Ki-67 protein, was increased gradually in NE (5%), hyperplasia (12%), CIS (30%), and IC (62%)u the differences were significant (P < 0.05) between CIS group and other groups., Conclusions: CX26 and CX43 may play important roles in carcinogenesis of cervical cancer. CXs may inhibit tumorigenesis and cell proliferation; it might be key factors in earlier stages when cellular malignant phenotypes have been established, or NE has been transformed into CIS.

Objective: To study the effect of Ar(+) laser on human vas deferens and to compare the effects of using different radiation levels with varying thickness of tissue and varying levels of injury., Methods: After initial tests on animals, four human scrotums were opened and treated directly with Ar(+) laser radiation. Then 58 human individual scrotums were treated with radiation by the method of trans-skin puncture. The rate of sperm reduction and elimination was tested., Results: In 60 cases, the sperms were found to be eliminated completely after six months of radiation treatment. In 2 cases the sperms were found not to be eliminated completely due to the insufficient radiation., Conclusion: Ar(+) laser is one of the best forms of radiation for coagulation of vas deferens. It can be used to coagulate vas deferens without any complications or sequelae.

Objective: To investigate the significance of detecting chimeric mRNA resulting from t(X;17)(p11.2;q25) in paraffin-embedded tumor tissues of alveolar soft part sarcoma (ASPS)., Methods: Formalin-fixed, paraffin-embedded tumor tissues from 8 cases of alveolar soft part sarcoma and 15 cases of controls (including 6 alveolar rhabdomyosarcomas, 6 renal cell carcinomas, 2 paragangliomas and 1 granular cell myoblastoma) were retrieved from the archival materials. ASPL-TFE3 fusion transcripts were analyzed in all samples by reverse transcriptase-polymerase chain reaction (RT-PCR). The quality of the mRNA was assessed using the house-keeping gene beta-actin., Results: ASPL-TFE3 fusion transcripts were detected in 6 of the 8 ASPS cases (4 being type 2 and 2 being type 1). The remaining 2 cases were negative for both beta-actin and ASPL-TFE3. No ASPL-TFE3 mRNA expression was detected in all the controls. PAX3/7-FKHR fusion transcripts were also detected in 4 of the 6 alveolar rhabdomyosarcoma samples., Conclusions: The expression of ASPL-TFE3 fusion transcripts in paraffin-embedded tumor tissues can serve as an useful molecular marker in the diagnosis of ASPS. It may also be helpful in elucidating the underlying pathogenesis of ASPS in subsequent retrospective studies.

Objective: To investigate the expression of the chimeric genes resulting from the specific chromosomal translocations in soft tissue sarcomas (STS) and its diagnostic significance for STS., Methods: The variety of fusion transcripts were detected in 103 cases of STS, including 30 cases of synovial sarcoma (SS), 15 cases of rhabdomyosarcoma (RMS), 25 cases of Ewing's sarcoma/peripheral primitive neuroectodermal tumors (ES/pPNET), 12 cases of dermatofibrosarcoma protuberans (DFSP), 14 cases of aveolar soft part sarcoma (ASPS), 3 cases of leiomyosarcoma (LMS), 2 cases of malignant fibrous histocytoma (MFH), and 2 cases of fibrosarcoma (FS); and 20 cases of control tumors by reverse transcription-polymerase chain reaction (RT-PCR) using formalin fixed, paraffin embedded specimens., Results: Of the 34 cases of SS 28 (93.3%) expressed SSX-SYT chimeric transcripts (14 were positive for SYT-SSX1, 9 for SYT-SSX2). Four of the six cases of alveolar RMS had a PAX3/PAX7-FKHR fusion transcript. None of the 9 cases of embryonic and polymorphic RMS expressed PAX3/PAX7-FKHR. Of the 25 cases of ES/pPNET, 19 were positive for EWS-FLI1 fusion transcript and 1 for EWS-ERG fusion transcript. COL1A1-PDGFB fusion transcript was expressed in 8 of the 12 cases (66.7%) of DFSP. Of the fourteen cases of ASPS, ten expressed ASPL-TFE3 fusion transcript. None of the 3 cases of LMS, 2 cases of MFH, 2 cases of FS, and 20 control cases contained any of the fusion transcript., Conclusion: Chimeric gene transcript resulting from specific chromosomal translocations is a reliable index for the molecular diagnosis of STS and RT-PCR assay for detection of specific fusion gene provides a useful tool for confirmation of the diagnosis of STS in diagnostically difficult cases and in retrospective studies.

Objective: To detect over-expression of AChR-gamma mRNA in rhabdomyosarcoma tissues by duplex RT-PCR and discuss its potential in diagnosis of rhabdomyosarcoma., Methods: Duplex RT-PCR was applied to the simultaneous detection of AChR-alpha and gamma subunit messenger RNA in 17 cases of rhabdomyosarcoma (9 ERMS, 6 ARMS, 2 PRMS). 20 cases of non-rhabdomyosarcomous small round cell tumors (6 poorly differentiated synovial sarcomas, 6 ES/PNET, 6 lymphomas, 2 neuroblastomas) and three normal muscle samples were also detected for AChR-alpha and gamma mRNA by the same method., Results: AChR-alpha and AChR-gamma mRNA were expressed in all the cases of rhabdomyosarcoma. The rate of quantity in both transcripts was AChR-gamma/AChR-alpha >or= 1, but the rate for three normal muscle samples was < 1. Cases of non-rhabdomyosarcomous small round cell tumors were all negative for AChR-gamma., Conclusion: AChR-gamma mRNA expression detected by molecular genetic methods is useful in diagnosis and differential diagnosis of rhabdomyosarcoma.