Background The retinal degeneration 11 (rd11) mouse is a newly discovered naturally occurring recessive animal model with lysophosphatidylcholine acyltransferase 1 (Lpcatl) mutation.Previous studies showed that the photoreceptor cells are characterized by typical rod-cone degeneration pattern in rd1 1 mice,while cone degeneration pattern in rd11 mice is unclcar.Objective Using immunofluorescence staining techniques with retinal wholemount,we aim to clarify the degeneration patterns of cone-function related M-opsin or S-opsin in different ages of rd1 1 mice.Methods A total of thirty rd1 1 and C57BL/6J mice at postnatal (P) day 14,28,42 (five in each age group) were sacrificed and retinal wholemounts were prepared.Immunohistochemistry was performed to identify the expression of M-opsin or S-opsin in retinal wholemounts,which were photographed with a fluorescent microscope.Cone opsins were compared between rd1 1 retinas and age-matched normal C57BL/6J retinas by manually counting the opsin positive cone cells in different quadrants of the retinas.Results The number of M-opsin or S-opsin positive fluorescent dots in each quadrant was similar at all ages of normal C57BL/6J retina.M-opsin positive fluorescent dots in dorsal/temporal,ventral/temporal,dorsal/nasal and ventral/nasal quadrants of rdl 1 retina at P28 were (414±32),(300± 8),(324 ± 22) and (250± 20)/0.037 mm2,which were lower than the age-matched normal C57BL/6J mice (t =4.114,15.225,7.505,17.990,all at P＜0.05).At the same time the S-opsin positive fluorescent dots in P28 rd11 were (8 ±4),(175 ± 16),(74 ± 13) and (315 ±20)/0.037 mm2,with significant decrease in comparison with those in the age-matched normal C57BL/6J mice (t =8.555,17.076,21.637,13.498,all at P＜0.05).With the development of retinal degeneration in rd11 mice,the M-opsin degeneration spread from central to ventral,nasal and then to temporal and dorsal peripheral retina;and the S-opsin loss started from dorsal/temporal to ventral/nasal retina.Conclusions Most of the M-opsin and S-opsins,especially the S-opsins in rd11 mice,degenerate in 6 weeks.Retinal wholemount and cone opsin immunofluorescent staining provide a useful tool to show the cone degeneration pattern and to evaluate the therapeutic efficiency in ongoing gene therapy study.

Background Trans-corneally subretinal injection in rodent model is a useful method for genetic therapy,stem cell transplantation and the study on the ophthalmic research.Standarized operation process is critical for the successful treatment.However,there is no literature to report the detailed procedure and the influence of this technique on morphology and function of retina.Objective This sudy was to introduce a method of trans-corneally subretinal injection and evaluate its influence on the morphology and function of retina.Methods Trans-corneallly subretinal injection was performed on the left eyes of 2-month-old SPF C57BL/6J mice after dilation of pupils.A 301/2G disposable needle was used to puncture the cornea within the pupil area near limbus and avoid touching the lens and irises under eye surgery microscope.Then,a 33G blunt needle was used to insert into the vitreous and toward subretinal space via corneal puncture.Normal saline with 0.1％ fluorescein sodium of 1 μl was slowly injected into the space,and 2.5％ hydroxypropyl methylcellulose was dropped on ocular surface for the observation of the fundus clearly.According to the percentage of the retina filled with subretinally injected solution,the experimental eyes were divided into 80％-100％ area group,50％-70％ area group after injection,and the mice in the pseudo-injected group,in which injection procedure stopped just before the solution was pushed in to the subretinal space did not inject any solution after punctured.The right uninjected eyes of the mice served as normal control group.Four eyes were selected for each group.The structural changes were evaluated by optical coherance tomograpby (OCT) 1 day,2 days,3 days and 5 weeks after injection,and retinal function was assessed by the recored of electroretinography (ERG) 5 weeks after injection.The retinal sepcimens were prepared to examin the morphological changes by hematoxylin and esosin staning.The use of care followd the Regulations for the Administration of Affair Concerning Experimental Animals of Zhejiang Province.Results About 70％ of the injected eyes showed that retinal blebs filled with injected green fluorescein solution occupied 50％ or more retinal area with minimal damages.The focal detachment between neurosensory retinal layer and retinal pigment epithelium (RPE) was exhibited 1 day postinjection,and almost all the retinas retached 2 days after injection.In the fifth week after injection,the amplitudes of ERG b wave were (386.25±37.88),(357.50±41.03),(324.25±53.45) and (410.50±14.88) μV in the sham operation group,50％-70％ area group,80％-100％ area group and normal control group,respectively,showing a significant difference among the 4 groups (F=3.574,P=0.047),and the amplitudes of b wave in the normal control group were higher than those in the 80％-100％ area group (all at P ＜ 0.05).The detachment between retinal neuroepithelium layer and RPE layer,cell proliferation and transposition in the outer nuclear layer were dispalyed under the light microscope in the sham operation group,50％-70％ area group and 80％-100％ area group,and the disordered outer segment of photoreceptors at the injecting area was seen in the 50％-70％ area and 80％-100％ area groups at five weeks after injection.However,retinal sructure and morphology were normal at the non-injection area.Conclusions Trans-corneally subretinal injection is an effective and safe way for subretinal injection.

Objective To analyze inducing factors of acute left heart failure(AHF) of emergency case.Methods According to different age and gender,the inducing factors of AHF of 211 patients were retrospectively analyzed.Results Infection(42.99%) was the most important inducing factors of AHF in all the four age-groups,but as for patients elder 80,the arrhythmias was also play an important part in inducing factors of AHF.The difference of inducing factors was found among the four age-groups.but the difference of inducing factors was not found between the two gender-groups. Conclusion Since the most important inducing factors of AHF of emergency case was infection,anti-infectives is important for preventing and controlling AHF.

Stargardt disease (STGD) is an inherited disorder of retinal pigment epithelium.Three genes have been found to be implicated in STGD including Abca4 (adenosine triphosphate-binding cassette,sub-family A,member 4),Elovl4 (elongation of very long chain fatty acids protein 4) and Prom1 (prominin-1).Target genes can be delivered to the retina by various methods such as lentivirus (LV) vectors,adeno-associated virus (AAV) vectors and non-viral nano-particles.The Abca4-/-,Elovl4-/-and Prom1-/ mice model are used to study the pathogenesis mechanism and treatment of STGD.Retinal function improved significantly upon gene therapy in these models.Based on these works using animal model,phase Ⅰ /Ⅱ a clinical trial of Abca4-associated STGD gene therapy are underway.As a LV vector,equine infectious anemia virus (EIAV) is used to carry the Abca4 gene.These studies will evaluate three dose levels of the EIAV vector for safety,tolerability and biological activity.Moreover,some preclinical attempts to deliver Abca4 via AAV have been made using a modified AAV vectors because of the large size of the ABCA4 cDNA.The good responses in animal models render STGD a very attractive object for human gene therapy after the successful of the phase Ⅰ /Ⅱ clinical trials of Leber's congenital amaurosis.

BACKGROUND:Transformation growth factor beta 1 is mostly used to induce the chondrogenic differentiation of bone marrow mesenchymal stem cel s, but there is a poor induction efficacy.
OBJECTIVE:To explore the chondrogenic differentiation of bone marrow mesenchymal stem cel s co-cultured with articular chondrocytes or induced by transforming growth factor beta 1.
METHODS:Articular chondrocytes and bone marrow mesenchymal stem cel s from SD rats were harvested and divided into 1:2, 2:1, 1:1 concentration groups. Cel s induced by transforming growth factor beta 1 acted as control group. After 20 days of induced culture, MTT was used to detect cel viability, alcian blue colorimetric assay was applied to measure glycosaminoglycan content, and western blot assay was employed to determine the expression of col agen type II.
RESULTS AND CONCLUSION:The absorbance value in the control group was significantly lower than that in the 1:1 and 2:1 groups (P<0.05). Glycosaminoglycan content and protein expression of col agen type II were also lower in the control group than the 1:2, 1:1, 2:1 groups. But there was no difference between 1:1 and 2:1 groups (P>0.05). The results show that bone marrow mesenchymal stem cel s co-cultured with articular chondrocytes can be induced to differentiate into chondrocytes, and meanwhile, there is a saturation phenomenon during the chondrogenic differentiation of bone marrow mesenchymal stem cel s.

Objective To investigate the CT findings of cardiac rhabdomyoma in childhood. Methods Five children with cardiac rhabdomyoma confirmed by operation was retrospectively analysed. Enhanced electron beam CT (EBCT) was performed in 4 children and enhanced 64-slice helical CT (MSCT) scan was performed in 1 infant. Three dimensional reconstruction was applied for the full display of the lesions based on the transverse section images. Results ( 1 ) Multiple masses or nodules originated from atrioventricular septum were observed in 4 children including 1 nodule from the anterior walls of the left and right ventricles. A mass was derived from the mitral valve in 1 infant (2) The CT value of the lesions after enhancement ranged from 80 to 180 HU and was similar to neighbouring left ventricular myocardium. (3) The ventricular outlet obstruction was found in 3 children including 1 infant with pericardial effusion and pulmonary inflamation of the right lower lobe. Conclusion The rhabdomyoma should be considered first when masses or nodules originated from atrioventricular septum and with the enhancement similar to neigbouring left ventricular myocardium in children.

Objective To learn whether plasmodium genetic attenuated sporozoites (GAS) can induce immunity against lung cancer ,in order to provide new ideas for the study of lung cancer vaccine .Methods Ther study was divided into two groups respec‐tively ,experimental group received intravenous injection of genetically attenuated sporozoites to immunize C57BL/6J mice and con‐trol group injection of phosphate buffer solution (PBS);after 14 days ,we subcutaneously inoculated lewis lung cancer (LLC) cells , calipers was used to measure tumor size .Immunohistochemical staining was detected tumor proliferation ,apoptosis ,and angiogene‐sis .Results There was statistically significant in tumor size .Immunohistochemical staining revealed that attenuated sporozoites in‐fection inhibited LLC eslls proliferation ,angiogenesis ,apoptosis .Conclusion The malaria attenuated sporozoites may provide a no‐vel strategy or therapeutic vaccine vector for anti‐lung cancer immune‐based therapy .

Background Retinitis pigmentosa (RP) is one of the causes of congenital blindness.It is well known that the degeneration process of rod cells is difficult to detect in RP.Retinal degeneration 12 (rd12) mice is a new,spontaneously arising mouse model for human Leber congenital amaurosis (LCA),and it is helpful for us to explore the pathogenesis and determine the treating target of RP.Objective This study was to investigate the natural disease process of short-length sensitive cone cells in rd12 mice,a LCA Rpe65rd12 (B6 [A]-Rpe65rd12/J) mouse.Methods The rd12 mice at postnatal (P) 14,P21,P35 and P90 were selected (5 mice for each),and the wild-type C57BL/6J (B6) mice with matched ages were included as controls.Photopic full-field electroretinogram (ERG) was recorded with Roland Q450SC UV visual physiology instrument.Cone response was recorded using single white light-emitting diode (LED) stimulation with the flash intensity of 1.00 cds/m2 and 1.96 cds/m2,and short wave-length sensitive cone response was recorded using ultraviolet light ([363 ±6] nm) stimulation with the flash intensity of 2.0 mWs/m2 and 3.0 mW/m2.The mice were sacrificed and retinal whole-mounts were prepared.The distribution and number of cone cells and UV-sensitive cone cells were detected by FITC-peanut agglutinin (FITC-PNA) and Cy3 immunofluorescence stainning,respectively.Results In P14 rd12 mice,the ERG responses of overall cone cells presented the negative waveform and the latency was delayed,and UV-sensitive cone response was unrecordable.The b-wave amplitude of overall cone cells reduced by 75％ in P21 rd12 mice compared with wild-type B6 mice,and the mean latency of b-wave in the P21 rd12 mice was significantly longer than that in the wild-type B6 mice ([102.80± 11.39] ms vs.[43.40± 5.60] ms) (t =-8.106,P =0.001).The mean b-wave amplitudes of U Vsensitive cone cells were (59.60± 36.00),(82.40± 12.22) and (68.43 ± 17.63) μV in the wild-type B6 mice,andthose in the rd12 mice were unrecordable.Immunofluorescence showed that a lager number of cone cells with green fluorescence were seen,and the expression of opsin with red fluorescence was displayed in the UV-sensitive cone cells of nasal lateral on retinal ventral side in P14 wild-type B6 mice;while only a few opsin positive-response cells were seen in P14,P21 and P35 rd12 mice.Conclusions In rd12 mice,the functional abnormality and quantitative reduction of cone cells appear in the early postnatal days,and the loss of UV-sensitive cone cells is earlier and more obvious.

Objective To observe the effect of simulated microgravity on the photopic negative response (PhNR) of full-field flash ERG in adult mice.Methods In an experimental study,forty-eight adult male C57BL/6J mice (48 eyes) were randomly divided into model and control groups.Model mice were further divided into three subgroups of 8 each:tail-suspended for 15 days (subgroup A),tail-suspended for 30 days (subgroup B),and tail-suspended for 30 days followed by returning to normal position for 30 days (subgroup C).The three control subgroups were similarly fixed with a harness but kept in the normal position for corresponding periods of 15,30,and 60 days.The mice were immediately examined using ERG-PhNR,flash VEP,OCT and visually-guided behavior in vivo,and subsequently sacrificed to analyze the retinal histology in vitro.PhNR amplitude was measured from baseline to PhNR trough.N1 peak-time and N1-P1 amplitude of VEP was analyzed.The escape duration was used to quantitatively evaluate the visual function of mice.In addition,inner retinal thickness was analyzed by OCT imaging.Data were compared by the independent sample t-test.Results PhNR amplitude in the model subgroup A was obviously lower than the corresponding control subgroup,the difference was statistically significant (t=-3.196,P＜0.01).There was no significant difference in PhNR amplitude between the model subgroup B or C and the corresponding control subgroup (t=-1.976,0.285;P＞0.05).There was no significant difference in FVEP N1 peak-time or N1-P1 amplitude between any of the three model subgroups and the corresponding control subgroup (P＞0.05).There was no significant difference in OCT-measured inner retinal thickness between any of the three model subgroups and the corresponding control subgroup (t=-0.461,2.073,-0.402;P＞0.05).The three model subgroups showed almost normal retinal structure,including the retinal ganglion cell,inner pexiform layer,inner nuclear layer,outer plexiform layer,outer nuclear layer,ellipsoid zone and RPE.There was no significant difference in visually-guided escape time between any of the three model subgroups and the corresponding control subgroup (t=-0.637,-0.955,1.297;P＞0.05).Conclusion Via tail-suspension,short-term simulated microgravity can affect the PhNR of flash ERG;however,the change is reversible and does not affect visual function of mice.

Esophageal cancer surgery originated in the early 20^th century. However, the true meaning of trans-thoracic esophagectomy and digestive tract reconstruction began in the 1930s. Almost at the same time, Japan and Western countries began the surgical exploration of esophageal cancer. Based on the pathological type of esophageal cancer in Asia, squamous cell carcinoma is the majority, and its biological characteristics and treatment strategies are different from those of European and American patients. After more than eighty years of development, the surgical treatment of esophageal cancer in Japan has been developed from the initial attempt, deep cultivation practice to the pursuit of excellence, and explored a set of more advanced surgical techniques and diagnostic strategies, which is unique in the world. On the basis of the establishment of the Japanese Society of Esophagus, Japanese scholars have developed and irregularly updated the Japanese Classification of Esophageal Cancer and published the professional academic journal Esophagus. The Japanese Clinical Oncology Group organized a number of phase Ⅲ clinical studies on esophageal cancer, providing strong evidence for the diagnosis and treatment of esophageal squamous carcinoma. Focused on the origin, development, current situation and future of esophageal cancer surgery in Japan, this paper summarized the development of esophageal cancer surgery in Japan through literature review, interviews with senior experts and Hot topics of esophageal cancer surgery-questionnaire survey of Japanese experts.