Analysis of interleukin-36 expression in tears of allogeneic hematopoietic stem cell transplantation patients and its correlation with the ocular surface microenvironment / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To detect the expression of interleukin(IL)-36(α, β, γ)in tears of patients undergoing allogeneic hematopoietic stem cell transplantation(allo-HSCT), investigate its correlation with ocular surface microenvironment, and further analyze the relationship between its expression and ocular graft-versus-host disease(oGVHD).METHODS: Prospective study. A total of 35 patients(70 eyes)underwent allo-HSCT in the hematology department of our hospital in January 2020 were selected, and 35 healthy volunteers(70 eyes)with appropriate age and gender were selected as normal control group. The patients in the allo-HSCT group were followed up 3 times after operation once every 3mo. The subjects with postoperative ocular symptoms were divided into oGVHD and Non-oGVHD group.Ocular surface disease index(OSDI)questionnaire, Schirmer test, tear break-up time(TBUT), corneal fluorescein staining(FL), and conjunctival impression cytology(CIC)was conducted in three groups. Furthermore, the expression levels of IL-36(α,β,γ)in tears were detected by ELISA.RESULTS: In the normal control group, IL-36(α, β, γ)expression levels were 74.32±5.27, 70.02±8.43, 97.41±8.66 pg/mL, respectively; in the allo-HSCT group, IL-36(α, β, γ)baseline expression levels were 77.27±7.03, 74.53±7.53, 100.77±9.74 pg/mL, with no statistically significant differences between the two groups(t=1.648, 1.954, 1.262, all P>0.05). There were no significant differences in IL-36α, IL-36β and IL-36γ in Non-oGVHD group at different time points(P>0.05), while there were significant differences in IL-36α, IL-36β and IL-36γ in oGVHD group at different time points(P<0.05). Compared with Non-oGVHD group, the levels of IL-36α and IL-36β at different time points were significantly increased in oGVHD group(all P<0.05).IL-36(α, β, γ)of oGVHD group was positively correlated with OSDI score, FL and CIC, while it was negatively correlated with TBUT and Schirmer test(all P<0.05).CONCLUSION: Evaluation of levels of tear IL-36(α, β, γ)can be of significance in diagnosing oGVHD after allo-HSCT. IL-36(α, β, γ)is highly expressed in the tears of oGVHD patients before the onset of ocular symptoms, and it is correlated with the ocular surface parameters.

Study on the correlation between tear film lipid layer thickness and macular microvascular changes in patients with diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To analyze the correlation between tear film lipid layer thickness(LLT)and macular microvascular parameters in diabetic retinopathy. <p>METHODS: Totally 60 eyes of type 2 diabetes patients with non-proliferative stage(NPDR group)and 60 eyes of proliferative stage(PDR group)with diabetic retinopathy diagnosed in our hospital from 2018-01/12 were selected, and 60 eyes of healthy volunteers with appropriate age and gender were selected as the normal control group. The tear film lipid layer thickness(LLT)was examined by Lipiview eye surface interferometer, while the foveal avascular zone(FAZ), superficial capillary layer(SCL)vessel density and deep capillary layer(DCL)vessel density were measured by optical coherence tomography angiography(OCTA)in three groups. The differences and correlations between the parameters were compared. <p>RESULTS: LLT(69.87±11.401nm)in normal control group was higher than that in NPDR(54.87±7.453nm)and PDR group(42.67±5.246nm), and FAZ(0.312±0.021mm2)was lower than that in NDPR group(0.389±0.037mm2)and PDR group(0.437±0.032mm2). The vascular density of SCL(51.977%±4.164%)was significantly higher than that of NPDR(47.067%±4.757%)and PDR(41.865%±5.512%), and that of DCL(49.578%±2.619%)was higher than that of NPDR(46.032%±2.622%)and PDR(40.598%±2.671%)(all <i>P</i><0.01). There was no correlation between LLT, FAZ, SCL and DCL in normal subjects. LLT was negatively correlated with FAZ in both NPDR group and PDR group(<i>r</i>=-0.922, <i>r</i>=-0.923, all <i>P</i><0.01), positively correlated with SCL(<i>r</i>=0.798, <i>r</i>=0.902, all <i>P</i><0.01), and had no correlation with DCL(<i>r</i>=0.140, <i>r</i>=0.073, <i>P</i>=0.285, <i>P</i>=0.581).<p>CONCLUSION: In diabetic retinopathy, the lipid layer of tear film is lower and the stability of tear film is decreased, and there is a correlation between diabetic dry eye and macular microvascular changes.

MiR-124-3p Enhances the Sansitivity of Chronic Myelogenous Leukemia Cell K562-R to Imatinib by Targeting ABCA2 / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect and mechanism of miR-124-3p-targeing regulating ABCA2 on chronic myelogenous leukemia cell K562-R.@*METHODS@#CML cells with miR-124-3p-overexpression and ABCA2-over-expression as well as subcutaneoustrans planted tumor nude mice were used as study objects. And the CML cells were divided into four groups: K562-R blank control, miR-124-3p mimic control, ABCA2-overexpression and mimic+PC ABCA2. The effects of miR-124-3p and ABCA2 on CML cells were analyzed. The levels of proliferation-, apoptosis- and autophagy- related protein were determined by Western blot. qRT-PCR was employed to detect the levels of miR-124-3p and ABCA2 in K562-R cells. The relationship between miR-124-3p and ABCA2 was validated by luciferase reporter system assays and bioinformatics. Hoechst/immunohistochemical staining and CCK-8 assay were performed to investigate the function involved.@*RESULTS@#miR-124-3p highly expressed in K562-S cells and lowly expressed in K562-R cells, however, ABCA2 lowly expressed in K562-S cells and highly expressed in K562-R cells. Over-expression of miR-124-3p significantly decreased ABCA2 level and cell growth, but increased autophagy and apoptosis in K562-R cells (P＜0.01). When ABCA2 was over-expressed, the K562-R cell growth was promoted and autophagy and apoptosis were inhibited (P＜0.01). The miR-124-3p promoted cell autophagy and apoptosis but inhibited cell growth in nude mice transplant tumor model (P＜0.01).@*CONCLUSION@#miR-124-3p can target ABCA2 to inhibit the growth of CML cells and promote the cell autophagy and apoptosis of CML cells.

Effect of Chlorambucil on the Apoptotic Signaling Pathway in Lymphoma Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study whether chlorambucil has apoptotic effect on the B cell lymphoma A20 cells and its exact mechanisms in apoptotic signaling pathway.</p><p><b>METHODS</b>The experimental cells were treated with 20 μmol/L chlorambucil, the control cells were treated with PBS. Annexin V-FITC Cell Apoptosis Detection Kit was used to examine cell apoptosis. Western blot was used to detect the expressions of active caspase-3, Survivin, NF-B and pAKT. Real-time fluorescent quantitative PCR was performed to examine the mRNA expression of Survivin.</p><p><b>RESULTS</b>Compared with the control group, the proportion of FITC/PI apoptotic cells and the expression of active caspase-3 (t=7.384, P=0.000) in the chlorambucil treatment group was significantly elevated. However, the expression of Survivin mRNA (t=4.384, P=0.000), protein expressions of survivin (t=12.360, P=0.000), NF-B (t=5.462, P=0.000) and pAKT (t=7.183, P=0.000) in the chlorambucil-treated group all significantly decreased.</p><p><b>CONCLUSION</b>The chlorambucil can induce the apoptosis of lymphoma cells, its mechanism may related with inhibition of PI3K/AKT signaling pathway, and expression of NF-B and survivin.</p>

Mutation Rate and Clinical Characteristics of CALR, JAK2 V617F and MPL W515K Genes in Patients With Primary Thrombocythemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyze the mutation rate and clinical characteristics of CALR, MPL W515K and JAK2 V617F genes in patients with primary thrombocythemia (PT).</p><p><b>METHODS</b>Fifty-six patients with PT were selected as the research objects in our hospital. The CALR and MPL W515K gene mutations were determined by genomic DNA-PCR direct sequencing of the PCR products, and the JAK2 V617F gene mutation was detected by allele specific PCR method.</p><p><b>RESULTS</b>Among the 56 patients with PT there were 14 cases of CALR gene mutation with the incidence rate of 25%, including 6 cases of type I, 5 cases of type II and 3 cases of type III. The sex, age, platelet(Plt) count, white blood cell (WBC) count and hemoglobin (Hb) level in the type I case of CALR gene mutation all were not significantly different from that in type II and III(all P>0.05); the WBC level in type III group significantly increased in comparison of type II group (P<0.05), while the sex, age, Hb and Plt levels showed no significant difference between the type III and type II groups (P>0.05). There were 3 cases of MPL W515K gene mutation with the incidence rate of 5.36%; 21 cases of JAK2 V617F gene mutation with the incidence rate of 37.50%. There were 13 cases of CALR gene mutation in negative patients with MPL W515K and JAK2 V617F (18 cases) with 72.22% incidence rate (13/18), and there was no cases of 1 or 2 gene mutations coexisted. The levels of Hb and WBC in peripheral blood of patients with CALR mutation were significantly lower than those of JAK2 V617F mutation (both P<0.05). In 56 cases, there were 3 cases of abnormal karyotype, with the incidence rate of 5.36%. The mutation rate of CALR gene in abnormal karyotypes (66.67%) was significantly higher than that of normal karyotypes (20.75%) (P<0.01).</p><p><b>CONCLUSION</b>The incidence of JAK2 V617F gene mutation increases in the patients with primary thrombocythemia; CALR mutation rate is higher in the patients with negative MPL W515K and JAK2 V617F gene mutation, which may closely correlate with abnormal karyotype; the levels of peripheral Hb and WBC in PT the patients with CALR gene mutation are significantly lower than those in patients with JAK2 V617F mutation.</p>

Safety and examination protocol optimization of fetal brain MRI / 西安交通大学学报(医学版)

With the emergence of ultrafast MR sequences,fetus MRI has been greatly improved.Because of its excellent detailed visualization and large view on soft tissue,MRI has more advantages in diagnosing fetal congenital brain malformation than ultrasound.Therefore,MRI has become an important examination method for prenatal screening and has a broad application prospect.This article reviews the safety,indications,scanning sequence, clinical application and precautions of fetal craniocerebral magnetic resonance.

Action Mechanism of Chlorambucil against Mantle Cell Lymphoma Cell Line Jeko-1 / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the action mechanism of chlorambucil against mantle cell lymphoma cell line Jeko-1.</p><p><b>METHODS</b>The effect of chlorambucil on Jeko-1 cell proliferation was measured by MTT method. The effect of chlorambucil on the apoptosis of Jeko-1 cell was detected by Hoechst staining and Annexin V-FITC dual staining. The activation of PI3K/AKT signaling pathway and the expression of BAX, BCL-2, procaspase 3, procaspase 8 and procaspase 9 were detected by Western blot.</p><p><b>RESULTS</b>0, 5, 10, 20 µmol/L chlorambucil could inhibit Jeko-1 cell proliferation at 24, 48, 72 h in a time- and dose-dependent manner. Chlorambucil of 0, 5, 10, 20 µmol/L increased the apoptotic rate of Jeko-1 cells, upregulated the expression of BAX, procaspase 3, procaspase 8, procaspase 9 and PI3K, increased the phosphorylation of AKT and down-regulated the expression of BCL-2.</p><p><b>CONCLUSION</b>The chlorambucil can induce the apoptosis of mantle cell lymphoma Jeko-1 cells via blocking PI3K/AKT signaling pathway.</p>

Amount Change of Peripheral Blood NK Cells in Patients with Hematologic Malignancies and Its Clinical Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the amount change of peripheral blood NK cells in patients with hematologic malignancies and its significance.</p><p><b>METHODS</b>A total of 200 patients with hematologic malignancies in our hospital from June 2013 to March 2015 were chosen as study objects, out of them 105 patients were in aute stage and 95 patients were in remisson stage. At same time 100 people from healthy medical examination in our hospital were chosen as control group. The mumber change and subgroups of their peripheral blood NK cells were analyzed and compared.</p><p><b>RESULTS</b>In control group the absolute number of NK cells was (412.91 ± 167.35)/µl, the relative number of NK cells was (13.31 ± 2.56) %; in group at acute stage of leukemia the absolute number of NK cells was (97.84 ± 23.18)/µl, the relative number of NK cells was (6.79 ± 0.78) %; in group at acute stage of lymphoma, the absolute number of NK cells was (101.79 ± 25.63)/µl, and the relative number of NK cells was (7.12 ± 1.03) %; in group at remission stage of leukemia, the absolute number was (297.17 ± 87.56)/µl, and the relative number was (10.15 ± 1.64) %; In group at remission of lymphoma, the absolute number of NK cells was (288.52 ± 118.52)/µl, and the relative number of NK cells was (10.82 ± 1.97) %. The number of NK cells between different groups showed statistical difference (P < 0.05). In remission group, the number of NK cells before and after treatment had statistical difference (P < 0.05). In control group, the number of CD56(bright) subgroup was (25.28 ± 4.72) %, the number of CD56(bright) subgroup at the acute stage of leukemia was (65.46 ± 11.21) %, and the number of CD56(bright) subgroup at the acute stage of lymphoma was (70.71 ± 12.14) %, the number of CD56(bright) subgroup at remission stage of leukemia was (23.35 ± 4.67) %, the number of CD56(bright) subgroup at remission stage of lymphoma was (24.89 ± 4.58) %. The number of CD56(bright) subgroup between different groups showed statistical significance (P < 0.05).</p><p><b>CONCLUSION</b>The number and function of peripheral blood NK cells in patients with hematologic malignancies have been confirmed to be obvious decrement, but after treatment the number of NK cells in those patients showed increment.</p>

Expression and Promoter CpG Island Methylation Status of miR-34b in Leukemia Cell Lines and Their Clinical Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the expression and promoter CpG island methylation status of miR-34b in leukemia cell lines and their clinical significance.</p><p><b>METHODS</b>A total of 10 cases of non-hematologic diseases were selected as control group, and the bone marrow cells of control group and HL-60, K562 cells were selected; the relative expression of miR-34b was detected in bone marrow cells, HL-60 and K562 cell lines by fluorescence quantitative PCR, and the MiR-34b methylation status was detected by methylation-specific PCR, the HL-60 and K562 cell lines were treated with decitabine, and the expression levels and methylation status of miR-34b in the 2 cell lines were detected by the same method. Has-miR-34b was transfected into K562 cells, which were divided into non-transfection group, negative control group and Has-miR-34b transfection group; if the transfection was successful, the cell proliferation should be recorded at different time points of culture, and the proliferation inhibition rate should be calculated.</p><p><b>RESULTS</b>The relative expression level of miR-34b in the control group was (5.23 ± 0.75), in HL-60 was (0.05 ± 0.01) and in K562 was (0.04 ± 0.01). The difference between 3 groups was statistically significant (F = 44.812, P < 0.01). The promoter regions of CpG island in HL-60 and K562 cell lines were methylated, while the bone marrow cells were not methylated in 10 cases of non hematologic diseases children.Through miR-34b expression levels of HL-60 and K562 cell lines significantly increased by decitabine treatment (P < 0.05), and the methylation of leukemia cell line promoter region CpG island was found before and after decitabine treatment, but after administration of decitabine the methylation significantly decreased, suggesting that decitabine has an inhibitory effect on methylation of promoter region CpG island. After being cultured for 48, 72, 96 and 120 hrs, the cell proliferation in Has-miR-34b transfection group reached to 24.8%, 46.7%, 33.6% and 4.7%, repectively, and significantly lower than that in non transfection group (P < 0.05).</p><p><b>CONCLUSION</b>CpG island methylation of miR-34b promoter region in leukemia cell lines can decrease the expression levels of miR-34b, which is also the reason why miR-34b can reduce the inhibition of cell proliferation, thus miR-34b might be a tumor suppressor gene involved in the regulation of leukemia.</p>

Effects of reactive nitrogen metabolites on NK cell-mediated killing of K562 cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the effects of the exogenous and endogenous reactive nitrogen metabolites (RNM) as NK cell inhibitors on NK cell-mediated killing of K562 cells and the influence of Tiopronin (TIP), glutamylcysteinylglycine (GSH) and histamine dihydrochloride (DHT) as RNM scavengers on reversing the suppressing effect of RNM.</p><p><b>METHODS</b>The exogenous ONOO(-) was administered in the NK+K562 culture system, then the RNM scavengers were added in the NK+K562+ONOO(-) culture system, respectively. The concentrations of RNM, TNF-beta and IFN-gamma, K562 cell inhibition rate (KIR) and the percentage of living NK cells were examined. IL-2+PHA were used as monocyte (MO) activators in the culture system of MO+NK+K562. Then TIP, GSH and DHT were administered and the parameters of NK cell activity were analyzed.</p><p><b>RESULTS</b>After exogenous ONOO(-) was administered in NK+K562 culture system, the percentage of living NK cells was decreased from (93.17 +/- 2.57)% to (71.87 +/- 1.02)% (P < 0.01) and KIR was decreased from (67.47 +/- 2.64)% to (43.44 +/- 2.87)% (P < 0.01). When TIP, GSH and DHT were administered into the systems, the percentage of living NK cells was increased to (91.13 +/- 3.67)% (P < 0.05), (88.03 +/- 1.46)% (P < 0.05), (73.60 +/- 2.76)% (P > 0.05), respectively; KIR was increased to (61.58 +/- 1.89)% (P < 0.05), (60.68 +/- 2.07)% (P < 0.05) and (45.26 +/- 3.31)% (P > 0.05), respectively. When IL-2/PHA were administered in the NK+K562+MO culture system, RNM products was increased from (82.10 +/- 6.60) micromom/L to (193.65 +/- 5.95) micromom/L(P < 0.01);KIR was decreased from (90.64 +/- 3.06)% to (61.29 +/- 2.22)% (P < 0.01). When the TIP, GSH and DHT were administered in the systems, RNM products were decreased to (91.32 +/- 6.81) micromom/L (P < 0.05), (84.66 +/- 5.99) micromom/L (P < 0.05) and (188.92 +/- 5.00) micromom/L (P > 0.05), respectively; KIR was increased to (84.31 +/- 4.56)%(P < 0.05), (81.65 +/- 3.09)% (P < 0.05) and (72.20 +/- 4.10)% (P < 0.05), respectively.</p><p><b>CONCLUSION</b>NK Cell-mediated killing of K562 cells can be suppressed by exogenous and endogenous RNM administration. Both of TIP and GSH can protect NK cells by scavenging RNM and enhance the antineoplasmic activity of NK cells.</p>

Pharmacokinetics of breviscapine liposomes following intravenous injection in Beagle dogs / 药学学报

<p><b>AIM</b>To prepare the breviscapine liposomes and study the pharmacokinetics of breviscapine liposomes in Beagle dogs.</p><p><b>METHODS</b>The cross-over design (two periods) was employed. Six Beagle dogs were administrated a single intravenous dosage of 28 mg of breviscapine liposomes and reference preparation, respectively, scutellarin in plasma of 6 dogs at different sampling time was determined by RP-HPLC. The pharmacokinetic parameters were calculated by 3P97 program and compared by statistic analysis.</p><p><b>RESULTS</b>The mean concentration-time curves of breviscapine liposomes and reference preparation were both fitted to two-compartment model with the main pharmacokinetic parameters as follows: T 1/2 alpha were (4.4 +/- 0.7) min and (1.8 +/- 1.3) min respectively; T 1/2 beta were (55 +/- 27) min and (28 +/- 23) min respectively; V(c) were (1 580 +/- 265) mL and (2 460 +/- 2 200) mL respectively; CL(s) were (88 +/- 10) mL x min(-1) and (324 +/- 69) mL x min(-1) respectively; and AUC(0-720) were (363 +/- 42) microg x min x mL(-1) and (102 +/- 19) microg x min x mL(-1) respectively. The T 1/2 alpha, CL(s) and AUC(0-720) of breviscapine liposomes all had significant difference from those of reference preparation, after the data were examined by a one-way analysis of variance (ANOVA).</p><p><b>CONCLUSION</b>Compared with the reference preparation, breviscapine liposomes had a much more higher concentration in plasma and contained characteristic of sustained-release, which ameliorated the pharmacokinetic properties of scutellarin.</p>

Ester prodrug of scutellarin: synthesis, physicochemical property and degradation / 药学学报

<p><b>AIM</b>To establish the fundamentals for the design of scutellarin prodrug and formulation with feasible physicochemical and biopharmaceutical properties by esterifying scutellarin, an active component with poor absorption extracted from Erigeron breviscapus of Chinese medicine.</p><p><b>METHODS</b>With the method of salifying followed by esterifying, ethyl and benzyl ester of scutellarin were synthesized. Glycolamide ester of scutellarin was also synthesized with an improved method. Their structures were confirmed by MS and 1H NMR. The solubility and partition coefficient of the prodrugs were determined and their degradations were investigated in various buffers and in human plasma. The emulsion and cyclodextrin complex of glycolamide ester were prepared and the protection of the ester from degradation was compared in the intestinal tract contents. Furthermore, the degradation of glycolamide ester in the homogenates of various intestinal segments was studied. Results Three prodrugs were synthesized successfully and their structures were confirmed. Glycolamide ester of scutellarin showed better stability in the aqueous solution (t(1/2) approximately =16 d, pH 4.2) and the shortest half-life in the human serum (t(1/2) approximately =7 min). Compared with scutellarin, the solubility of glycolamide ester was increased about ten times in pH 4.0 buffer, and about thirty five times in water. Partition coefficient of the glycolamide ester increased significantly from -2.56 to 1.48. However, the ester degradation in the homogenates of intestinal mucus would be an obstacle for its absorption. The degradation rates were in the order duodenum > ileum > or = jejunum > colon. The emulsion showed a better protection of glycolamide ester from the degradation than cyclodextrin complex.</p><p><b>CONCLUSION</b>Glycolamide ester of scutellarin shows better physicochemical properties than ethyl and benzyl eater of scutellarin, but its stability in intestinal tract needs to be improved. The emulsion or / and colon-targeted delivery may be selected as one of strategies to decrease the presystemic degradation.</p>

Effect of reduced glutathione as anti-leukemic immune adjuvant / 中国实验血液学杂志

To investigate the reversal effect of reduced glutathione (GSH) on suppression of NK cells by reactive oxygen metabolites (ROM) in K562 cells, interleukin-2 (IL-2) or mononuclear cell (Mo) was added in cultured cell line of K562 cells and NK cells, the yield of ROM and K562 cell suppression rate were observed. Then the histamine dihydrochloride (DHT) or GSH was added in the mixed cultured cell lines, the ROM production and K562 cell suppression rate were observed. The results showed that the ROM yield increased from 33.17 +/- 5.08 U/L to 223.59 +/- 9.41 U/L by IL-2, and K562 cell suppression rate increased from 65.56% to 85.89% by IL-2 (P < 0.01). The ROM yields were 389.79 +/- 43.83 U/ml, 456.74 +/- 42.77 U/ml and 601.42 +/- 21.92 U/ml respectively, and K562 cell suppression rates were 82.36%, 81.36% and 48.09% respectively, when Mo was added in the mixed cultured cell lines under ratios of E/Mo being 10/2, 10/5 and 10/10. When E/Mo was 10/2, DHT or GSH was added in the mixed cultured cell line ROM yield decreased from 389.79 +/- 3.83 U/L to 50.21 +/- 2.4 U/L or -3.58 +/- 9.49 U/L (P < 0.05) respectively. With increase of concentration of DHT or GSH, the ROM yield in the mixed cultured cell line decreased (P < 0.05), the K562 cell suppression rate increased from 82.53% to 94.64% or 96.39% (P < 0.05), the more ROM yield, the less K562 suppression rate (P < 0.05). When E/Mo is 10/5 or 10/10, the ROM yield decreased by the high concentration of DHT or GSH (P < 0.05), but the K562 cell suppression rate not increased by every concentration of DHT or GSH. GSH was as effective as DHT in the reversing ROM and increasing K562 cell suppression rate. It is concluded that GSH may reverse ROM and increase K562 cell suppression rate, and GSH is as effective as DHT, but GSH has less side-effect than DHT. Therefore, GSH would be better antileukemia immune adjuvant.

Pharmacokinetics of breviscapine and its beta-cyclodextrin complex in rats / 药学学报

<p><b>AIM</b>To establish RP-HPLC method for determination of plasma scutellarin concentration and study of the pharmacokinetic behavior of scutellarin in rat after ig administration of breviscapine and its beta-cyclodextrin complex (breviscapine-beta-CD).</p><p><b>METHODS</b>Mobile phase composed of methanol and acetate buffer. The column was Shim-pack C18. Twelve rats randomized into 2 groups were separately given breviscapine and breviscapine-beta-CD at single dose of 10.8 mg.kg(-1). Drug in plasma was extracted and determined by HPLC. The pharmacokinetic parameters were calculated by 3P97 software.</p><p><b>RESULTS</b>Linearity was obtained over the range of 10-400 ng.mL(-1). The recovery was 95.32%-98.81%. C(max) and AUC(0-12 h) of breviscapine were (154 +/- 18) ng.mL(-1) and (710 +/- 126) ng.h.mL(-1). For breviscapine-beta-CD, C(max) and AUC(0-12 h) were (328 +/- 31) ng.mL(-1) and (1,093 +/- 200) ng.h.mL(-1), respectively. There were significant differences of AUC(0-12 h) between breviscapine and breviscapine-beta-CD (P < 0.01).</p><p><b>CONCLUSION</b>The assay method was suitable for the determination of scutellarin plasma concentration in rat. Brevescapine-beta-CD showed greater absorption compared with that of brevescapine.</p>

Scavenger of reactive oxygen metabolites reverses the ROM induced inhibition of NK cell-mediated killing effect on K562 cell in vitro / 中国实验血液学杂志

To investigate the effect of a new reactive oxygen metabolites (ROM) scavenger as immune adjuvant in NK cell-mediated killing effect on K562 cell, IL-2 and PHA were used to activate monocyte to produce ROM, and different concentrations of tiopronin as ROM scavenger was used in the cultivated systems with different ratio of monocytes plus NK cells and K562 cells, while histamine dihydrochloride (DHT) with different concentrations was used as positive control. The reuslts indicated that after IL-2 and PHA were supplemented in the cultivated systems mixing with NK cells and K562 cells as the E/T ratio was 10/1, the ROM production increased from 33.17 +/- 25.02 U/ml to 223.59 +/- 59.41 U/ml (P < 0.05) while K562 cell inhibition rate (KIR) increased from 65.56% to 85.89% (P < 0.05). When the monocytes as the E/MO ratios of 10/2, 10/5 and 10/10 were supplemented respectively, ROM production increased correspondingly (ROM production was 389.79 +/- 43.83 U/ml, 456.74 +/- 42.77 U/ml, 601.42 +/- 21.92 U/ml, respectively), and KIR was on the other round (KIR was 82.36%, 81.36%, 48.09% respectively). Tiopronin, DHT were used in the K562 + NK + MO + IL-2/PHA cultivated systems as the E/MO ratio was 10/2, the ROM production also decreased from 389.79 +/- 43.83 U/ml to -1.20 +/- 60.70 U/ml, 50.21 +/- 22.4 U/ml (P < 0.05), respectively, however KIR increased from 82.53% to 96.09% and 94.64% either (P < 0.05). Higher concentrations of tiopronin and DHT were used, ROM production decreased accordingly. There showed a reverse correlation between ROM production and KIR (r = -0.518). When E/MO ratio was 10/5 or 10/10, tiopronin at any testing concentration and DHT at the higher testing concentration could reduce the ROM production (P < 0.05), but did not improve KIR significantly (P > 0.05). Tiopronin was as good as DHT in ameliorating KIR (P > 0.05) and better than DHT in scavenging ROM (P < 0.05). It is concluded that (1) Monocytes are the major resources of ROM, and the ROM derived from monocytes can disable NK cells in killing neoplasm cells (K562 cells); (2) A new ROM scavenger, tiopronin, can scavenge ROM effectively, and reverse the ROM induced inhibition of NK cell-mediated killing of K562 cell in a certain extent. And tiopronin is better than DHT in scavenging ROM, and as good as DHT in up-regulating KIR. The new ROM scavenger tiopronin with less side effect may take the place of DHT as adjuvant during the adoptive immuno-therapy in leukemia.

In vitro and in vivo stability of 9-nitrocamptothecin lactone form in rats / 药学学报

<p><b>AIM</b>To investigate the in vitro and in vivo stability of 9-nitrocamptothecin lactone form in rat plasma.</p><p><b>METHODS</b>The specific and accurate HPLC method was developed for quantifying 9-nitrocamptothecin lactone form and the total lactone and carboxylate forms simultaneously. By using of this method, the ratios of lactone form to the total in rat plasma at different time were determined in vitro and in vivo. The results were compared to determine which was the main factor influencing the stability of 9-nitrocamptothecin lactone form in rat plasma in vivo.</p><p><b>RESULTS</b>The stability of lactone form in rat plasma was much higher in vivo than that in vitro.</p><p><b>CONCLUSION</b>Blood cells help to increase the stability of 9-nitrocamptothecin lactone form. Clearance from blood in vivo is the primary factor which influences the plasma stability of 9-nitrocamptothecin lactone form. The kinetic process of 9-nitrocamptothecin lactone form and total drug in rats were both best fitted to a two-compartment model. However, the process of 9-nitrocamptothecin carboxylate form in vivo was best fitted to a one-compartment model.</p>

Mechanisms of action of transportation of liposomes and chitosan-coated liposomes containing leuprolide across intestine and Caco-2 cell / 药学学报

<p><b>AIM</b>To investigate the mechanisms of action of transportation of liposomes and chitosan-coated liposomes containing leuprolide across rat intestine and Caco-2 cell.</p><p><b>METHODS</b>Everted-gut technique and Caco-2 cell were used to study the transport properties of free leuprolide, liposomes and chitosan-coated liposomes containing leuprolide. Caco-2 cell was used to study the effect of chitosan concentration and the order of addition on the permeation of liposomes.</p><p><b>RESULTS</b>The transport of leuprolide was passive diffusion. Probably because the entrapment by liposomes prevents the transport of leuprolide across the rat intestine and Caco-2 cell, the permeation amount of leuprolide from liposomes was lower than that of the free drug. However, liposomes protected the leuprolide from degradation. Chitosan promoted the transport of leuprolide from liposomes and there was no obvious difference in enhancement effect from the concentration of 0.1% to 0.5%. On the other hand, the incubation of chitosan with liposomes may weak the enhancement effect of chitosan.</p><p><b>CONCLUSION</b>Chitosan-coated liposomes showed both protection and enhancement effect, therefore, they may promote the oral absorption of leuprolide.</p>

Relationship of matrix metalloproteinase-9 and ripening of pregnant cervix / 第三军医大学学报

Objective　To investigate whether matrix metalloproteinase-9 (MMP-9) was synthesized in pregnant cervix during parturition and its source and distribution. Methods　Cervical species (n=10, each weighing about 0.3 g) were taken from pregnant women immediately after delivery. Other cervical species (n=7) were served as negative control from those non-pregnant women but undergoing uterotomy due to other benign diseases. Immunohistochemical method (ABC) was carried out to detect the expression of MMP-9, with a monoclonal antibody against MMP-9. Results　Positive staining of MMP-9 was found in the cytoplasm of polymorphonuclear leukocytes (PMN) that had infiltrated into cervix or located in blood vessels of cervix. Scattered light positive staining were found in some interstitial cells of the cervix. No other cells including fibrocytes and lymphocytes were positive to MMP-9. No positive staining was found in control tissues. Conclusion　There are strong expressions of MMP-9 in pregnant cervix in term labor, derived mainly from infiltrated PMN. MMP-9 may be an important regulator in the process of cervical ripening.

Relationship of matrix metalloproteinase-9 and ripening of pregnant cervix / 第三军医大学学报

Objective　To investigate whether matrix metalloproteinase-9 (MMP-9) was synthesized in pregnant cervix during parturition and its source and distribution. Methods　Cervical species (n=10, each weighing about 0.3 g) were taken from pregnant women immediately after delivery. Other cervical species (n=7) were served as negative control from those non-pregnant women but undergoing uterotomy due to other benign diseases. Immunohistochemical method (ABC) was carried out to detect the expression of MMP-9, with a monoclonal antibody against MMP-9. Results　Positive staining of MMP-9 was found in the cytoplasm of polymorphonuclear leukocytes (PMN) that had infiltrated into cervix or located in blood vessels of cervix. Scattered light positive staining were found in some interstitial cells of the cervix. No other cells including fibrocytes and lymphocytes were positive to MMP-9. No positive staining was found in control tissues. Conclusion　There are strong expressions of MMP-9 in pregnant cervix in term labor, derived mainly from infiltrated PMN. MMP-9 may be an important regulator in the process of cervical ripening.