Objective To study the clinic value of digital three-dimensional shaping titanium mesh in repairing skull defects and reconstructing temporal muscle attachment points. Methods Eighty-two patients with skull defects from December 2013 to January 2016 were enrolled in this study. They were divided into two groups according to patients' will with 41 patients in each group. The patients in control group received traditional hand-shaping titanium mesh repair, the patients in observation group received individual digital forming technology to produce titanium mesh repair and reconstruct the temporal muscle attachment points at the titanium mesh temporal corresponding position. The rate of appearance satisfaction, operation time, the number of titanium nail and the rate of postsurgery adverse reaction was compared between the two groups. Results The rate of appearance satisfaction in observation group was significantly higher than that in control group: 97.56%(40/41) vs. 80.49%(33/41), P<0.05. The operation time and the number of titanium nail and in observation group was significantly lower than that in control group:(8.9 ± 1.6) h vs. (12.4 ± 2.4) h, (10.3 ± 0.9) pieces vs. (11.8 ± 1.8) pieces, P < 0.01. The rate of postsurgery adverse reaction in observation group was significantly lower than that in control group: 0 vs. 9.8%(4/41), χ2 = 4.21, P < 0.05. Conclusions The operation of repairing skull defects and reconstructing temporal muscle attachment points with digital three-dimensional shaping titanium mesh is simple. It can shorten the operation time, decrease the postsurgery adverse reaction, and increase the appearance satisfaction.

Objective To establish an in vivo imaging method of normal or tumorous liver in mice by using a new type nanoparticle contrast agent, ExiTron nano 12000, coupled with micro-CT imaging.Methods Six 6-8-week old male C57BL/6J mice were randomly divided into group A and group group B, by intravenous injection of 50μL and 100μL Ex-iTron nano 12000, respectively.In vivo Micro-CT scans were performed before contrast agent injection, 3 minutes, 24 hours, 7, 14, 28 and 56 days after injection.To determine which dose is suitable for long-term studies, gray scale value a-nalysis was performed on selected region of interest ( ROI) in the left lobe and right anterior lobe of the liver, and the chan-ges of liver tissue contrast was monitored after ExiTron nano 12000 injection.Three male HBV transgenic mice bearing liver tumors ( group C) were intravenously injected with the determined dose of ExiTron nano 12000 and were monitored by mi-cro-CT scans as above described.At 56 days after ExiTron nano 12000 injection, the mice were sacrificed and liver sam-ples were taken for histological analysis.Results Cross-sectional images taken at various time points and the average gray scale value ( AGSV) analysis in the mouse liver revealed that the AGSV peaked at 24 hours after injection of contrast rea-gent and good contrast still presented in the livers within 56 days of observation for both groups, though group B showed a significantly higher contrast than group A (P＜0.01).Those data indicated that the dose of group B (100μL) was better to maintain ExiTron nano 12000 in the liver of mice for a long time.Contrast-enhanced by 100μL of ExiTron nano 12000, the liver tumor nodules in the mice of group C could be clearly delineated by Micro CT imaging during a 56 days observa-tion.Histological analysis revealed atypical hyperplasia, enlarged nuclei with hyperchromasia and cell necrosis in the tumors.Conclusions An in vivo imaging method was established to non-invasively visualize mouse liver using micro-CT combined with nanoparticle-based contrast agent and this technology may be applied to a live imaging of murine primary liv-er tumors.

Object To establish an induced mouse model of type 2 diabetes mellitus, and compare it with db/db mouse model of spontaneous type 2 diabetes.To evaluate the two mouse models objectively, and provide an experimental basis for the choice of animal model and its practical application in diabetes studies.Methods A mouse model of induced type 2 diabetes was established by feeding C57BL/6J mice with high-fat and high-sugar diet for four weeks and taking daily intraperitoneal injections of streptozotocin ( STZ) for consecutive 3 days.Four weeks after infection, the gross appearance of kidney and liver of the mice was assessed, glucose tolerance was tested, serum biochemical indices and expression of se-rum IL-2, IL-4, IL-6, IFN-γ, TNF-α, IL-17, and IL-10 were assayed, and were compared with those of the db/db mouse models of spontaneous type 2 diabetes.Results Obvious differences were found in the kidneys and liver gross appearance of the two types of mouse models and the control group.Both the two groups showed significant differences in the blood glu-cose levels at each time point (P＜0.05) and low glucose tolerance function, but there were no significant differences in blood glucose levels of the two types of mouse models.Compared with the control group, the serum biochemical indices GLU, GHOL and LDLC of the two types of mouse models were significantly increased (P＜0.05).Meanwhile, the blood lipid level of the mouse model of induced type 2 diabetes was higher than that of the db/db mouse models of spontaneous type 2 diabetes.In comparison of immune indices, except IL-2,the serum cytokine levels of the two types of mouse models were significantly higher than those of the control group (P＜0.05).Moreover, the serum cytokine levels of db/db mice were higher than those in the mouse models of induced type 2 diabetes, and the IL-6, IFN-γ, TNF-αalso had obvious differences.Conclusions Both the two types of mouse models of type 2 diabetes successfully simulate the human diabetes to some extent, but there are still certain differences according to different etiology of diabetes.We would suggest that peo-ple may take our data as reference and chose appropriate mouse models according to the requirement of their research.

ObjectiveTo compare three types of acute Mycobacterium tuberculosis infection mouse models established through different infection routes and to set up the theoretical basis for further developing,selecting and applying these animal model in the tuberculosis-related research.MethodsStandard strain of Tubercle bacillus H37Rv was diluted to 1 × 106 colony forming unit (cfu)/mL.The mice were infected with the bacteria through different routes including intravenous injection,intranasal administration and inhalation of bacteria aerosol.Six weeks after the infection,the mice were euthaniz ed and necropsied. The lung tissues were collected and gross changes were observed.The colony counting was performed and the lung tissues were assessed by HE staining,acid fast staining.The e xpression level of tumor necrosis factor (TNF)-α per unit area in lung tissue was detected by immunohistochemistry. The data were analyzed by t test. Results The amounts of Mycobacterium tuberculosis in lung tissues of mice in inhalation group,intranasal administration group and intravenous injection group were (6.290±0.028),(6.150±0.021) and (6.120±0.008) lg cfu/mL,respectively; while no Mycobacterium tuberculosis was detected in control group. The difference between infection group and control group was statistically significant (t =3.762,P＜0.01),while there were no significant differences among infection groups with different infection routes (P＞0.05).According to the results of gross observations and histological assessment,the pathological changes were observed and red tubercle bacillus was detected by acid-fast staining in the lung tissues of all the mice in infection group.The results of immunohistochemistry showed that the expression levels of TNF-α per unit area were as follows:intravenous injection group (0.049 × 106 )＜intranasal administration group(0.759×106) ＜ inhalationgroup(1.042×106), whichwere statistically different (t =2.504,P＜ 0.05).ConclusionInhalation of bacteria aerosol may be the most efficient method to establish tuberculosis infection mouse model compared to intravenous injection and intranasal administration.

Objective To established a bacteriemia model of BALB/c mice after infection with a ST-239 methicillin-resistant staphylococcus aureus(MRSA) strain, which was isolated and identified from Shanghai Huashan Hospital. Methods We monitored the clinical signs and gross observations of MRSA-infected mice, and examined the histopathology among different groups. Results This isolated MRSA strain ST-239 can induced a typical bacteriemia in BALB/c mice, including the severe mortality and extensive histopathologic injury. However, higher survival rate and slight inflammatory injury were observed in vancomycin-treated mices. Conclusion The solid results obatined in this model will benefit us to study the pathogenic characteristics and patholgenesis in MRSA-induced bacteriemia, and propeled us to seek a safety cure approaches in the future.

Objective To compare and analyze the differences and characteristics of three strain mouse models in-fected by influenza virus aerosol inhalation, and provide the reference for choosing the appropriate infection model in the re-search of pathogenesis of influenza and the development of vaccines and drugs.Method C57BL/6, BALB/c and ICR mice were infected with A/Puerto Rico/8/34 (H1N1) virus strain by aerosol inhalation.The symptoms and body weight of mice were observed every day.At 3, 7, 14 days after infection, the mice were sacrificed.The lungs of mice were weighed, then virus assay and pathological observation were carried out.Results The three strains of mice were infected.The sur-vival rate in the C57BL/6 mice was lower than those in the BALB/c and ICR mice.The lung index and viral load of C57BL/6 mice were significantly higher than those of ICR mice ( P<0.05) at 3 days after infection.The pathological changes of C57BL/6 mice were also more obvious than other two strains.Compared with other two mouse strains, the weight recovery of BALB/c mice was the slowest.The survival rate in BALB/c mice was higher than that of C57BL/6 mice and lower than that of ICR mice.The lung index and viral load were not significantly different among the three strains of in-fected mice.The pathological changes among the three strains of infected mice were similar, but the degrees of pathological changes in the BALB/c mice were milder than in the C57BL/6 mice and worse than in the ICR mice.Compared with other two mouse strains, the process of disease is similar, but the body weight, mortality, lung index, viral load, and the micro-scopic pathological changes were lighter in the ICR mice than in the other two strain mice.Conclusions The three strain mouse models can be established by influenza virus aerosol inhalation, but showing different characteristics.Appropriate strain mice can be chosen to build model according to different research purpose in the experiment.

Objective To observe the role of resting metabolic rate (RMR) in evaluation of animal model of glucocorticoid-induced Kidney Yin deficiency and Kidney Yang deficiency syndrome.Methods Male BALB/c mice were divided into control group,model group,Jinkuishenqi pill group,and Zhibaidihuang pill group.The later 3 groups were given drinking water containing corticosterone (first dissolved in 1% ethanol,with a final concentration of corticosterone 100 μg/mL).The control group was given drinking water containing 1% ethanol.RMR was measured by closed fluid pressure respirometer.At the end of the experiment,the mice were sacrificed to detect the weight index of perirenal fat,epididymal fat,quadriceps,and tibialis anterior muscle.ELISA assay was used to detect the level of serum hormones.Histological changes of the liver and kidney were examined by HE staining.Malondialdehyde (MDA) content,succinate dehydrogenase (SDH) activity,cytochrome c oxidase (COX) activity and ATP level were measured.Results Compared with the control group,the RMR of model group was significantly increased at the 2nd day of beginning of the experiment,reached the highest on the 4th day (P<0.01),then decreased gradually,and to the 66th day,the RMR was significantly reduced (P<0.05).Use of corticosterone resulted in decrease of the serum levels of thyroxine (T4),muscle mass index,SDH activity,COX activity and ATP level,while increase of fat mass index and MDA level.The two Kidney nourishing prescriptions reduced animal mortality,and reduced the content of MDA in liver tissue.But only Jinkuishenqi pill increased the RMR at the 4th and 66th days (P<0.05),and significantly improved the liver SDH activity,COX activity and ATP level (P<0.01).The Zhibaidihuang pill showed no such effects.Conclusions RMR can be used for evaluation of animal model of Kidney Yin or Kidney Yang deficiency induced by glucocorticoids.

Objective To establish uPA inducible expression system using recombinant retroviral system for the further construction of inducible uPA-SCID animal model .Methods The Inducible expression system need to construct two plasmids:pLNHXO1O2-Alb-GLUC-FMN2A -rtTA and pLNHXO5O6-TRE2-uPA-IRES-ZsGreen respectively. Both plasmids were based on retroviral vector pLNHX , Albumin promoter gene ( Alb) and rtTA gene or uPA gene and ZsGreen were obtained by PCR reaction and inserted into pLNHX .The Gaussia enzyme fluorescent element ( GLUC) was used to monitor rtTA expression in pLNHXO1O2-Alb-GLUC-FMN2A-rtTA, and the ZsGreen for uPA expression monitoring in pLNHXO5O6-TRE2-uPA-IRES-ZsGreen.The correct constructed plasmids were transfected into packaging cell line GP 2-293 to gain recombinant viral particles .NIH/3T3 cells were infected with these viral particles and selected with G 418.Gene expression in the surviving cells was confirmed by the PCR method .Results The recombinant retroviral vectors harbouring target genes were successfully cloned .The rtTA gene in pLNHXO1O2-Alb-GLUC-FMN2A-rtTA was expressed, and uPA can be induced to express in pLNHXO5O6-TRE2-uPA-IRES-ZsGreen by doxycycline (Dox) when the plasmid transfected into the HepG-Tet-on cell.The constructed recombinant two retroviral vectors were transfected into GP 2-293 packaging cells respectively to gain infectious viral particles .Then,NIH/3T3 cells were infected with these viral particles and single-cell clones which stably expressed the transgenes were successfully established .Conclusion This study primarily established uPA inducible expression system , it laid a foundation for the murine model of inducible liver damage , and provided a novel technical platform for further building the liver humanised murine models for viral hepatitis studying .

ObjectiveTo investigate the effect of acup point injection and rehabilitation techniques on recovery of upper limb function and activities of daily life (ADL) in patients with shoulder-hand syndrome (SHS) after cerebral infarction.Methods Ninety-six patients with SHS after cerebral infarction admitted to Jiuquan City Traditional Chinese Medicine Hospital and Jiuquan City People's Hospital from January 2014 to February 2016 were enrolled, and they were randomly assigned to observation group (56 cases) and control group (40 cases). The patients in the two groups were given the same routine comprehensive cerebral infarction treatments and rehabilitation technological training. In the control group, additionally intermediate frequency physiotherapy was used, while in the observation group, acupuncture and acupoint injection of salvia miltiorrhiza and ligustrazine were added. The patients in two groups were treated consecutively for one month, afterwards, the upper limb motor function, ADL, shoulder joint pain, degree of swelling and range of motion were assessed, and the clinical effects were observed.Results After treatment, the modified Fugl-Meyer motor function score (FMA), modified barthel index (MBI) and the ranges of voluntary activities of shoulder abduction, adduction, flexion, extension, internal rotation and external rotation in patients of two groups were significantly increased than those before treatment; the pain visual analog scale (VAS) and the edema volume difference in observation group were lower than those in the control group, and after treatment, the changes of above indexes were more significant in the observation group than those in the control group [FMA: 70.10±6.38 vs. 41.76±18.69, VAS: 2.00±1.37 vs. 5.00±1.25, MBI: 75.17±3.51 vs. 51.56±2.12, edema volume difference (cm3): 2.13±2.05 vs. 5.75±1.17, flexion: (153.1±6.7)° vs. (56.0±10.9)°, extension: (39.6±5.8)° vs. (17.2±6.7)°, abduction:(168.1±9.6)° vs. (52.1±8.9)°, adduction: (52.1±3.6)° vs. (21.8±4.3)°, external rotation: (49.8±7.6)° vs. (23.2±5.2)°, internal rotation: (107.0±8.5)° vs. (51.2±6.1)°], the total effect in observation group was significantly higher than that in the control group [91.1% (52/56) vs. 70.0% (28/40),P < 0.05].Conclusions Acupuncture and acupoint injection combined with three-level therapeutic regimen of rehabilitation technological training for treatment of shoulder-hand syndrome after cerebral infarction can reduce the patient's pain and edema, and improve upper limb motor function and activities of daily living.

Objective To establish a model for cutaneous squamous cell carcinoma by irradiation of SKH-1 hairless mice with solar-simulated ultraviolet (solar UV), and to explore the biological characteristics of the model. Methods A total of 91 SKH-1 hairless mice were randomly divided into seven experimental groups (n = 10) and seven control groups (n = 3). The mice in experimental groups were irradiated with minimal erythema dose of solar UV 4 times per week for various durations (4, 8, 12, 16, 20, 24, 28 weeks), while the control mice received no irradiation. The general status and skin appearance of mice were observed during the treatment process. Mice were killed immediately after the last irradiation at different time points and pathological examination was carried out to observe the histological changes of skin lesions. Results Papules measuring equal to or more than 1 mm in diameter began to develop in some mice in experimental group 10 weeks after the first irradiation; tumors began to appear in 39.3% (11/28) of the remaining mice in experimental group on week 20, and in 100% (10/10) of the remaining mice on week 28. The cumulative dose approximated to 26.99 J/cm2 for UVB and 242.91 J/cm2 for UVA after 28-week irradiation. No tumor was observed in the control mice. Pathological examination revealed characteristic changes of squamous cell carcinoma in 30% of the mice on week 12, 33.3% on week 16, 60% on week 20, 87% on week 24, and 100% on week 28. Conclusions Ultraviolet could induce the hyperplasia of skin in SKH-1 hairless mice, and even cause the development of cutaneous squamous cell carcinoma after prolonged irradiation.