Study on the effect of Pin1 novel depressor all-transretinoic acid in collagen-induced arthritis mice / 中华风湿病学杂志

Objective To investigate the effect of Pin1 novel depressor all-transretinoic acid (ATRA) in collagen-induced arthritis (CIA) mice and elucidate the underlying mechanisms.Methods CIA model mice were established,then the qualified ones were treated with ATRA and other medicines respectively.Paw thickness and volume were measured once a week in each group.The production of tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) was detected by enzyme-linked immunosorbent assay (ELISA) in serum.Pathological changes of joints tissues were observed by hematoxylin and eosin (HE) staining.Western blot was used to detect the expression of Pin1 and nuclear factor-κB (NF-κB) between different groups.The measurement date were compared with single factor analysis of variance and independent sample t test.Results Compared with healthy control group,the paw thickness,volume and arthritis scores were significantly increased in CIA mice,compared with disease control group,joints swelling and arthritic score index were reduced in both treatment groups;TNF-α and IL-6 expression were significantly increased in CIA mice [(50.1±1.3) pg/ml,(67.6±8.3)) pg/ml] when compare with health control group [(47.1±0.6) pg/ml,(43.0± 5.2) pg/ml] (t=-9.0,P＜0.01;t=-6.9,P＜0.01),and both were decreased in treatment groups compare with disease control group (TNF-α,F=35.5,P＜0.05;IL-6,F=17.15,P＜0.01);HE staining study demonstrated that ATRA and Dexamethasone could significantly suppressthe pathological changes of joints tissues;Western blot assay demonstrated that the higher arthritis scores was,the more synovial pin1 expressed in mice,moreover ATRA and Dexamethasone decreased Pin1 and NF-κB expression in CIA mice synovial.Conclusion ATRA can successfully decrease inflammation scores and joints pathology damage in CIA mice via weaken the synovial Pin1 expression.

Mechanism of all-transretinoic acid increasing retinoblastoma sensitivity to vincristine

Objective: To explore the mechanism of all-transretinoic acid (ATRA) increasing retinoblastoma (RB) sensitivity to vincristine, and the inhibiting effect of vincristine combined with ATRA treatment on the SO-RB50 cell proliferation. Methods: SO-RB50 cells were cultivated by routine culture method. Different concentrations of vincristine or ATRA were added into culture solution. After 48 h, cell counting kit-8 was used to detect the median inhibitory concentration (IC

Mechanism of all-transretinoic acid increasing retinoblastoma sensitivity to vincristine

OBJECTIVE@#To explore the mechanism of all-transretinoic acid (ATRA) increasing retinoblastoma (RB) sensitivity to vincristine, and the inhibiting effect of vincristine combined with ATRA treatment on the SO-RB50 cell proliferation.@*METHODS@#SO-RB50 cells were cultivated by routine culture method. Different concentrations of vincristine or ATRA were added into culture solution. After 48 h, cell counting kit-8 was used to detect the median inhibitory concentration (IC50) of vincristine combined with ATRT treatment to SO-RB50 cells. SO-RB50 cells were divided into drug combination group, vincristine group, ATRA group and control group. Different drugs were added into the culture solution respectively for cell culture based on the IC50 value. Cell counting kit-8 was used to detect the cell proliferation every 24-h cultivation. After continuous determination for 6 d, data was processed to draw the cell growth curve. After drug use for 72 h, flow cytometry was used to detect the proportion of different cell cycles of SO-RB50 cells in each group. After drug use for 48 h, annexin V/propidium iodide method was used to detect the SO-RB50 cell apoptosis in each group.@*RESULTS@#The IC50 value of vincristine treatment on the SO-RB50 cells was 0.11 μmol/L, and ATRT was 12.84 μmol/L. The cell growth curve in control group rose gradually along with the extended culture time, but after vincristine and ATRA treatment, the cell growth curve was smooth and steady. The cell increment was the least in drug combination group and its cell growth curve was the smoothest. There was significant difference in A450 48 h and 72 h after treatment (Fgrouping = 77.316, P < 0.001; Ftime = 86.985, P < 0.001). Compared with control group, A450 value in drug combination group, vincristine group, ATRA group was significant lower (P < 0.001). Compared with control group, the G2/M phase cell proportion in vincristine group was significantly increased, while the G0/G1 phase cell proportion was significantly decreased; the G0/G1 phase cell proportion in ATRA group was significantly increased, while the S phase cell proportion was significantly decreased (FG0/G1 = 85.878, Fs = 56.455, FG2/M = 85.878, P < 0.001). After 48 h, there was significant difference in SO-RB50 cell apoptosis rate among groups (F = 11.312, P < 0.05). The apoptosis rate in drug combination group was significantly higher than that of other groups (P < 0.001).@*CONCLUSIONS@#ATRA can increase the sensitivity of SO-RB50 cells to vincristine. Vincristine combined with ATRA treatment can significantly increase the inhibiting effect on SO-RB50 cells, which may be related with promoting cell apoptosis and involving in cell cycle control.

All-transretinoic acid combined with arsenious acid and chemotherapy in the treatment of acute promyelocytic leukemia patients with 10 years for survival / 中国基层医药

Objective To study the curative effect and characteristics of all -transretinoic acid (ATRA) and arsenic acid (As2 O3 )combination chemotherapy for acute promyelocytic leukemia (APL).Methods ATRA (30 -60mg/d),As2 O3 (0.1%As2 O3 d1 -28),chemotherapy(anthracycline -based chemotherapy regimens and homoharringtonine -based chemotherapy regimens or high -doses of cytosine arabinoside and small -doses of metho-trexate)were used alternately 4 -5 cycles.Results 12 cases had complete response(CR),CR rate was 100%,first cases and relapse cases all got CR,the median time needed 30 days to get CR.Follow -up deadline,all cases 10 year disease -free survival (DFS )rate was 100%,all reexamination minimal residual disease (MRD)was negative. Conclusion Using this method to treat adult APL,the CR rate is high,has less adverse reaction,and did not produce resistance phenomenon.It can be used for APL consolidation and maintenance therapy.This is one of the ways that can be recommended in the treatment.

Isolation and culture of rat limbal stem cells and research on trans differation capacity / 局解手术学杂志

Objective To isolate and culture the rat limbal stem cells ( LSCs) in vitro,and investigate briefly their capacity for transdif-ferentiation into neural stem cells ( NSCs) with cytokine EGF,bFGF and RA.Methods LSCs derived from rats were cultured and identified by immunohistochemistry in vitro.LSCs were induced to differentiate into NSCs in the presence of EGF (20 ng/mL) ,bFGF (10 ng/mL) and with or without of RA(group 1 or group 2)(25 ng/mL) for seven days.Cultures without factors were used as control group.Then the neural marker Nestin of the coultured cells were measured by immunohistology staining.Furthermore,the positive cell rate was counted under micro-scope between the 2 groups and analyzed by statistical software.Results It showed that P63 was positive in LSCs.Nestin in both of the dif-ferentiation groups was positive at the rate of (77.01 ±6.32)%and (84.01 ±5.43)%respectively,of which the second group was higher than the first one (P<0.05).However,it was negative in the control group.A band of Nestin protein from cells was detected by western blot assay.Conclusion LSCs are successfully isolated and cultured in vitro.LSCs could be induced to differentiate into NSCs in the presence of EGF and bFGF.Moreover,the differentiation capability is enhanced in the condition of RA.

Effects of All-transretinoic Acid on Cell Apoptosis and p53 Expression in Rabbits' Carotid Atherosclerosis / 中国康复理论与实践

@#ObjectiveTo observe the effects of all-transretinoic acid (ATRA) on cell apoptosis and p53 expression in rabbits' carotid atherosclerosis. Methods40 New Zealand rabbits were randomly divided 4 groups: normal group (NOR), control group (CON), surgery group (SUR) and treat group (TRE). Each group was fed with high cholesterol forage. The SUR and TRE groups were made up carotid atherosclerotic model by air desiccation injury after 2 weeks. The TRE group was fed with ATRA before surgery. On the 2nd and 4th week after operation, the target vessel was taken out. The intima hyperplasia was observed by microscopy through HE staining. Cell apoptosis was determined by TUNEL staining and p53 was observed by immunohistochemistry staining. ResultsOn the 4th week, the area of neointima in SUR, TRE and CON were (0.389±0.021) mm2, (0.113±0.038) mm2, (0.069±0.016) mm2 (P<0.01). The positive cells rate of TUNEL (8.40±0.45)% and p53 (3.02±0.38)% in TRE were higher than those of SUR (8.59±0.42)% and (2.23±0.29)%, respectively (P<0.01). ConclusionATRA may induce cell apoptosis in atherosclerotic plaque and inhibit the proliferation of intima associated with p53 in vessel injury.

Inhibit the proliferation of leukemia cell line NB_4-R2 with all-transretinoic acid-resistant in vitro by bortezomib / 白血病·淋巴瘤

Objective To investigate the effect of bortezomib on the proliferation and apoptosis in leukemia cell line NB_4-R2 in vitro and provide some new evidences for the treatment of acute promyelocytic leukemia APL with ATRA-resistant using bortezomib. Methods NB_4-R2 cells were incubated with bortezomib at different does for 48 h. The proliferation capacity was measured by MTT assay, the morphology of cell apoptosis observed with Hoechst33342 staining by fluorescence microscopy and the percentage of apoptosis calculated by flow cytometry. The expression of apoptosis protein of cleaved (poly ADP-ribose polymerase, PARP) and Caspase-3 were determined by Western blotting. Results The proliferation of NB_4-R2 cells were obviously inhibited by bortezomib in vivo and the role of inhibition was a does-dependant manner within the scope of the bortezomib concentration from 1-5 μg /L.The incidence of inhibition was up to 74.9 % at the bortezomib concentration of 5 μg/L. Within this scope of the bortezomib concentration mentioned above, the role of inhibition of proliferation of NB_4-R2 cells mainly showed an increase of the late apoptosis, and the percentage of apoptosis was up to 78.9 %. In the meaning time, the expressions of the apoptotic protein of cleaved PARP and Caspase-3 were up-regulated in NB_4-R2 cells after treated with bortezomib by Western blotting assay. Conclusion Bortezomib can inhibit the proliferation of NB_4-R2 cells in vivo by inducing cell apoptosis.

Treatment of acute promyelocytic leukemia: A single institution experience / Tratamiento de la leucemia promielocítica aguda: Experiencia de una sola institución

The results of the treatment of 14 patients with promyelocytic leukemia (PML) treated with all trans-retinoic acid (ATRA), combined chemotherapy (CT) and prophylactic prednisone are reported; the median age was 30 years (range 7 - 49). A complete remission (CR) was obtained in 13 / 14 patients (93%). All patients were given ATRA fully as outpatients; the CR was achieved after the administration of ATRA in five patients, whereas in the remaining eight, CT was required to achieve it. There were no instances of the ATRA syndrome. One patient relapsed with a PML/RAR-a negative PML 575 days after achieving the CR, failed to respond again to ATRA and died. The median overall (OS) and disease free survival (DFS) has not been reached, being above 4,000 days, whereas the 12-month DFS was 93%, the three and five years DFS being 85%. The treatment employed differs from others in: Oral prednisone is used prophylactically, ATRA is given on an outpatient basis and adriamycin is used instead of other anthracyclines. The results are similar to those obtained in other centers worldwide and it is possible that the prophylactic administration of prednisone precluded the development of the full-blown ATRA syndrome in this group of patients.

Se informan los resultados del tratamiento en una sola institución de 14 pacientes con leucemia aguda promielocítica (LAPM) en quienes se empleó la combinación de ácido holotrans-retinoico (ATRA) quimioterapia combinada y prednisona profiláctica. La mediana de edad fue de 30 años (rango 7-49). Se obtuvo remisión completa (hematológica y molecular) (RC) en 13 pacientes (93%); a todos los pacientes se les administró el ATRA de manera ambulatoria. La RC se obtuvo con el ATRA en cinco pacientes; en los demás la RC se obtuvo después de habérseles administrado la quimioterapia con citarabina/adriamicina. No hubo ningún caso de síndrome de ATRA. Un paciente recayó con una LAPM PML/ RAR-a negativa, 575 días después de haber logrado la RC y falleció. Otro paciente recayó 20 meses después de haber logrado la RC y fue rescatado con el mismo esquema de tratamiento; permanece en segunda remisión molecular por más de seis años. La mediana de supervivencia (SV), tanto global como libre de recaídas de todo el grupo, no se ha alcanzado y es mayor de 4,000 días, en tanto que la SV a 12 meses fue de 93% y a tres y cinco años de 85%. El esquema de tratamiento usado difiere de otros en que se usa prednisona oral, se administra el ATRA de manera ambulatoria y se usa adriamicina y no otras antracidinas; los resultados son similares a los obtenidos con otros esquemas parecidos en otros sitios del mundo; es posible que el uso profiláctico de prednisona haya eliminado la ocurrencia del síndrome de ATRA.