Regulatory Effect of Liuwei Dihuangtang and Guifu Dihuangtang on Low-dose Hydrocortisone Reduction and Discontinuance in Mice with Drug-induced Deficiency Syndrome / 中国实验方剂学杂志

Objective:To study the effect of the kidney-tonifying recipes on mouse syndrome and adrenal function induced by low-dose hydrocortisone reduction and discontinuation. Method:The 48 male ICR mice were randomly divided into normal control group, hydrocortisone group, Liuwei Dihuangtang group and Guifu Dihuangtang group, with 12 rats in each group. Each group was given 1.32 mg·kg-1·d-1 hydrocortisone for 28 days, and half a dose for 7 days, and the administration was discontinued 14 days before modeling. In the reduction and discontinuation phase, 12.5 g·kg-1·d-1 Liuwei Dihuangtang and 13.5 g·kg-1·d-1 Guifu Dihuangtang were simultaneously administered to the corresponding group for 21 d. On the 28th and 49th day of the experiment, the characteristic information of mice was detected by the experimental methodology of syndrome differentiation and treatment in mice (holding power, body surface infrared temperature). The mice were sacrificed on the 50th day of the experiment, the spleen and thymus were separated, and the spleen and thymus index were calculated, serum corticosterone content is measured by enzyme-linked immunosorbent assay (ELISA), the total RNA of the adrenal gland was extracted, and the expressions of Star, cholesterol-side-chain cleavage enzyme (Cyp11a1), Cyp21a1, Cyp11b1, low-density lipoprotein receptor (Ldlr), scavenger receptor class B type I (Scarb1/SRB1), Hmgcr, acyl-CoA-cholesterol acyltransferase-1 (Acat1), hormone sensitive lipase (Lipe/HSL), insulin-inducible gene 1 (Insig1) and sterol regulatory element binding transcription factor 2 (Srebf2) were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). Western blot was used to detect the expressions of LDLR, SRB1, CYP11A1 and cytochrome P450 family 21 subfamily A member 2 (CYP21A2) in adrenal gland. Result:Compared with the normal control group, the body weight and the body mean infrared temperature of the model group were significantly decreased after 28 days of continuous administration (P<0.01). After the administration of the drug, the mice in the model group showed a significant decrease in the grip strength (P<0.05), a significant increase in the thymus index (P<0.01), a significant decrease in the serum corticosterone content (P<0.05). Expressions of adrenal Cyp21a1 and Hmgcr genes decreased (P<0.05, P<0.01), Lipe and Acat1 gene expressions increased significantly (P<0.01), adrenal CYP11A1, SRBI protein expressions decreased significantly, and LDLR protein expression increased. Compared with the model group, the body weight of Guifu Dihuangtang group decreased significantly after 21 days of treatment with traditional Chinese medicine (P<0.01), the spleen index of Liuwei Dihuangtang group decreased significantly (P<0.01), expressions of Cyp11a1, Cyp21a1, Acat1 and Lipe genes in Guifu Dihuangtang group were significantly increased (P<0.05, P<0.01), expressions of Ldlr and Scarb1 genes were significantly decreased (P<0.05), and expressions of Ldlr, Acat1 and Lipe genes in Liuwei Dihuangtang group were significantly decreased (P<0.05, P<0.01), and expressions of CYP11A1 and SRBI proteins in Liuwei Dihuangtang and Guifu Dihuangtang increased. Conclusion:Guifu Dihuangtang can promote the recovery of adrenal function in mice with drug-induced deficiency syndrome by correcting expression of adrenal cortex synthase.

Clinical observation of a triple procedure of stimultaneous penetrating keratoplasty combined with extracapsular cataract extraction and intraocular lens implantation / 眼科新进展

Objective To evaluate the operative outcomes of a triple procedure including simultaneous penetration keratoplasty (PKP),extracapsular cataract extraction (ECCE) and intraocular lens (IOL) implantation,and to investigate the relationship between postoperative corneal refractive power and preoperative lens diopter.Methods This retrospective analysis study involved 15 patients who had undergone a triple procedure surgery in Beijing You'an hospital from April to October 2016.Outcomes including the best corrected visual acuity (BCVA),intraocular pressure (IOP),corneal refractive power,axial length,postoperative complications,corneal endothelial cell counts and the survival of corneal graft were determined one year after surgery.Results All corneal grafts were transparent and corneal endothelium were (1974.20 ±472.82) cell · mm-2.The mean postoperative LogMAR visual acuity (0.80 ±0.27) had a significant improvement compared with the mean preoperative LogMAR visual acuity (2.63 ±0.62) (t =13.042,P ＜0.001).There were no statistically significant differences in preoperative IOP (15.27 ± 2.37) mmHg (1 kPa =7.5 mmHg) and postoperative data (14.53 ± 3.04) mmHg (t =0.685,P =0.505),preoperative axial length (23.69 ±2.01) mm and postoperative data (23.62 ±2.12)mm (t =-0.138,P=0.893)and preoperative keratometry (45.01 ± 1.66) D of the control eye and postoperative data (42.56 ± 5.48) D (t =1.202,P =0.260).The postoperative spherical equivalent refractive was (0.40 ±4.65) D,and the target refraction was (0.58 ±0.25)D.Conclusion The triple procedures are safe and effective for the treatment of patients with coexisting corneal pathologies and cataracts.Selection of emmetropia lens diopter may result in the satisfactory postoperative visual acuity.However,unpredictable postoperative corneal curvature changes will still affect the final refractive state.

Research Progress in PTEN as Target Spot in TCM for Prevention and Treatment of Primary Liver Cancer / 中国中医药信息杂志

Phosphatase and tensin homolog deleted on chromosome ten (PTEN) is an important negative regulatory molecule in the development and progression of primary liver cancer and other cancers. TCM has unique advantages in the prevention and treatment of primary liver cancer. This article reviewed the research progress in PTEN as target spot in TCM for prevention and treatment of primary liver cancer in the past 10 years, and provided references for further research and clinical application.

Effect of Osthole on Adrenocortical Function in Y1 Mouse Adrenocortical Tumor Cells / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of osthole (Ost) on adrenocortical function in Y1 mouse adrenocortical tumor cells.</p><p><b>METHODS</b>Y1 mouse adrenocortical tumor cells were taken as subjects in this experiment. In 10.0%, 1.0%, and 0.1% serum DMEM-F12 medium, Y1 cells were treated with 1, 10, 25, 50, 100, and 200 micromol/L Ost for 24 and 48 h. 0.1% Dimethyl Sulfoxide (DMSO) was taken as negative control group and 1 mmol/L (Bu) 2cAMP as positive control group. Cell growth morphology was observed under inverted microscope. Contents of corticosterone were tested by ELISA. Expression levels of steroids synthase such as Star, Cyp11a1, Cyp21a1, Hsd3b2, Cyp11b1, Cyp11b2, Cyp17a1, and Hsd17b3 mRNA were detected by Real time quantitative PCR (RT-qPCR).</p><p><b>RESULTS</b>Y1 cell proliferation was obviously inhibited by 100 and 200 micromol/L Ost, and its inhibitory effect was more significant in 0.1% serum medium. Compared with the negative control group, gene expressions of Star, Cyp11a1 , Cyp21a1, Hsd3b2, Cyp11b1, Cyp17a1, and Hsd17b3 were significantly enhanced in the posi- tive control group (P < 0.05). Y1 cell corticosterone levels significantly increased in 50 micromol/L Ost treatment group after 24-and 48-h intervention (P < 0.05). Contents of corticosterone increased more obviously in 25 and 50 +/- mol/L Ost treatment groups after 48-h intervention, as compared with 24-h intervention (P < 0.01). After 24-h intervention, expression levels of Star, Cyp21a1, and Hsd3b2 genes were significantly up-regulated in 25 and 50 lLmol/L Ost groups (P < 0.05). Star gene expression was further enhanced after 48-h intervention (P < 0.05). However, Ost showed no effect on Cyp11a1 (P > 0.05). Additionally, gene expressions of Cyp11b1 and Cyp17a1 were significantly enhanced by 10, 25, and 50 pLmolIL Ost after treatment for 24 and 48 h (P < 0.05). Ost showed no obvious effect on Cyp11b2 and Hsd17b3 expressions.</p><p><b>CONCLUSION</b>Ost could regulate adrenal cortex function and promote corticosterone synthesis and secretion through strengthening gene expressions of steroidogenic enzymes.</p>

A single institution, retrospective study of treatment experience in primary mediastinal germ cell tumors: elucidating the significance of systemic chemotherapy / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Primary malignant germ cell tumors (GCTs) of mediastinum are rare neoplasms. We introduce our institutional experience in managing patients with primary malignant GCTs of the mediastinum, focusing on the analysis of therapeutic modalities.</p><p><b>METHODS</b>A retrospective review was done in 39 consecutive patients with mediastinal malignant GCTs treated in our institution between 1991 and 2007.</p><p><b>RESULTS</b>A total of 39 patients were enrolled in this study with a median age of 27 years. The 5-year overall survival (OS) and progression-free survival (PFS) rates of the whole population were 60.2% and 57.7%, respectively. Stratified by the histology, 18 patients (46.2%) had seminoma and 21 patients (53.8%) had nonseminomatous germ cell tumors (NSGCTs). The 5-year OS rate of patients with seminoma was 87.4% as compared with 36.7% in patients with NSGCTs (P = 0.0004). The 5-year PFS rate was also significantly higher in seminoma patients (87.4% vs. 31.6%, P = 0.003). For 19 patients with NSGCTs managed with multi-modality treatment, chemotherapy exposure appeared to impact the prognosis. The 5-year OS rate was 44.9% in patients with chemotherapy exposure as compared with 20.0% in patients without it (P = 0.43).</p><p><b>CONCLUSION</b>Our study confirmed the significance of systemic chemotherapy in the treatment of primary mediastinal GCTs.</p>

The correlation between adrenal corticosteroids and cytokines expressions in mice of different syndromes / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the cytokines expressions in the adrenal gland and its correlation with serum adrenal corticosteroids in mice of different syndromes.</p><p><b>METHODS</b>Using the quantitative four diagnosis and syndrome differentiation methods, 60 normal mice and 190 H22 liver cancer bearing mice were syndrome typed. Serum corticosterone and aldosterone were tested by ELISA, and mRNA expressions of cytokines in the adrenal gland were detected using Real-time PCR.</p><p><b>RESULTS</b>Mice of different syndromes were obtained, such as normal mice of no syndrome, normal mice of vigorous qi syndrome, normal mice of qi deficiency syndrome, liver cancer bearing mice of excessive evil toxic syndrome, liver cancer bearing mice of evil lying in the middle syndrome, liver cancer bearing mice of weak evil toxic syndrome, and liver cancer bearing mice of poisonous pathogenic factors and qi deficiency syndrome. The serum corticosteroids were significantly higher in the liver cancer bearing mice than in the normal mice (P < 0.05). The cortex hormones increased most significantly in the liver cancer bearing mice of excessive evil toxic syndrome (P < 0.05). Compared with the normal mice, IL-1beta, IL-2, IL-6, IL-10, IL-12alpha, IL-12beta, and TNF-alpha gene expressions increased in the liver cancer bearing mice, while only expressions of IL-1alpha and IL-5 decreased. But the expressions of IL-13 and transforming growth factor beta1 (TGF-beta1) showed no regularity. The expressions of IL-4 and INF-alpha were not detected in all mice. It is notable that the more severe degree of poisonous pathogenic factors, the higher the expressions of serum corticosterone and aldosterone levels as well as IL-6, the lower expressions of IL-1beta, IL-2, IL-5, IL-12alpha, IL-12beta, and TNF-alpha.</p><p><b>CONCLUSIONS</b>The increased serum corticosteroid level in liver cancer bearing mice could possibly be induced by chronic tumor stress, partial cytokines were involved in the synthesis and secretion of the adrenal hormone. Of them, IL-6 might positively regulate the secretion of corticosteroids, while IL-1beta, IL-2, IL-5, IL-12alpha, IL-12beta, and TNF-alpha might negatively regulate their secretions.</p>

Analysis of the dynamic changes of blood hormone levels in H22 liver cancer mice of poisonous pathogenic factors syndromes to different degrees / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the dynamic changes of blood hormone levels in H22 liver cancer mice of poisonous pathogenic factors syndromes (PPFS) to different degrees.</p><p><b>METHODS</b>Two hundred and twenty mice were injected with H22 tumor cells from their armpits. On the ninth day after inoculation the mice of severe poisonous pathogenic factors syndrome (SPPFS) and of mild poisonous pathogenic factors syndrome (MPPFS) were screened. Besides, another normal control group consisting of 30 mice was set up. The mice were killed on the tenth and eleventh day after inoculation (as the 1st and 2nd time window). The weight of the tumor, the wet weight of the thymus and the spleen were weighed. The plasma adrenocorticotropic hormone (ACTH), corticosterone, aldosterone, thyroid hormone T3 and T4, testosterone, TNF-alpha, and IFN-gamma were detected by ELISA. All the aforesaid laboratory parameters were analyzed.</p><p><b>RESULTS</b>The tumor weight was obviously larger in mice of the SPPFS group than in those of the MPPFS group at the same time window (P < or = 0.05). Compared with the normal control group, the thymus was obviously atrophied (P < or = 0.05), the spleen was significantly enlarged (P < or = 0.05), the plasma ACTH significantly increased (P < or = 0.05) in the SPPFS group at the two time windows. But the increment of ACTH was less in the MPPFS group. The plasma corticosterone showed similar tendency as that of ACTH. At the 1st time window the plasma testosterone significantly increased in the two groups (P < or = 0.05). The plasma testosterone and T4 showed a decreasing tendency in the SPPFS group. The plasma TNF-alpha and IFN-gamma levels showed an increasing trend in the two groups. Correlation study showed that the degree of PPFS was negatively correlated with qi deficiency (r = -0.766, P < or = 0.05) and T4 (r = -0.738, P < or = 0.05). The degrees of PPFS was positively correlated with the plasma ACTH level (r = 0.635, P < or = 0.05). The degree of qi deficiency was positively correlated with yang heat syndrome (r = 0.632, P < or = 0.05). The plasma ACTH was negatively correlated with T4 (r = -0.504, P < or = 0.05). The plasma testosterone was positively correlated with TNF-alpha (r = 0.619, P < or = 0.05).</p><p><b>CONCLUSIONS</b>PPFS occurs naturally and shows difference to different degrees in the development of H22 liver cancer. The disorders of neuroendocrine hormones and the suppression of the immune function show dynamic changing trends.</p>

Characteristics of gene expression of adrenal cortical steroid synthetase and its regulatory factor in mice with H22 liver cancer of different patterns / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the characteristics of gene expression of adrenal cortical steroid synthetase and its regulatory factor in mice with H22 liver cancer of different patterns.</p><p><b>METHODS</b>Syndromes revealed in mice with H22 tumor were differentiated by quantified four diagnostic methods and syndrome differentiation, and mice with commonly encountered patterns (A: evil-toxin accumulation pattern, B: qi-deficiency pattern, C: yang-qi deficiency pattern and D: qi-yin-yang deficiency pattern) were screened out for subjecting to the study. Two batches of GeneChip Mouse Exon 1.0 ST Array detection were performed in the selected mice for detecting the gene expressions of adrenal cortical steroid synthetase and its regulatory factor, with the analysis performed put stress on the differential expressions in mice of various syndrome patterns.</p><p><b>RESULTS</b>Data obtained from the two batches detection showed well repeatability, in which similar genes of high or low expression emerged. The adrenal cortical steroid synthetase genes, such as Cyp11a1, Star, Cyp11b2, Cyp21a1, Hsd3b and Hsd17b were highly expressed, with few difference among the four patterns. However, Cyp11a1 was down-regulated and Cyp1b2 up-regulated in all patterns; Hsd3b1 and Cyp21a1 down-regulated in pattern A and B, but up-regulated in pattern C and D. As for the expressions of the relative regulatory factors, Cyb5b and Wnt4 were down-regulated but Fdx1, Fdxr, Hsd11b1, Por, Agt and Nr 0b1 were up-regulated in all patterns; Nr5al down-regulated in pattern A but up-regulated in other three patterns; Nr4al and Nr4a2 up-regulated in pattern A and down-regulated in the others.</p><p><b>CONCLUSIONS</b>The adrenal cortical steroid synthetase genes are rather conservative and stable in mice bearing H22 liver cancer, part of the expression might be correlated to the condition of disease and essence of syndromes, embodying the differences among different patterns in the same disease.</p>

A method of acutely isolating rat dorsal root ganglion neurons for patch-clamp study of single-channel / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To establish a method of acutely isolating dorsal root ganglion (DRG) neurons for patch clamp study of single-channel.</p><p><b>METHODS</b>DRG neurons of rats were acutely isolated by enzymatic digestion and mechanical blowing.</p><p><b>RESULTS</b>The acutely isolated DRG cells were easy to form the higher sealing resistance (> 5G Omega), which lowered noise level, so that pA-level single channel currents could be recorded.</p><p><b>CONCLUSION</b>The acutely isolated DRG neurons in this study are an ideal for patch-clamp study of single-channel.</p>

Expression of Pax6 gene in retinoblastoma / 眼科

Objective To detect the gene expression of Pax6 in retinoblastoma (Rb).Design Experimental study.Participants Six cases of fresh Rb tumors and two established Rb cell lines.Six cases of retina tissues as normal control group.Methods Pax6 gene ex- pression was assessed in two established Rb cell lines,six primary tumors and six cases of retina tissues using the reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot.Main Outcome Measures Expression of Pax6 gene in fresh retinoblastoma tis- sue and cell lines.Results The mRNA of Pax6 gene was positive in all two established Rb cell lines and the six primary Rb tumors using RT-PCR.The protein of Pax6 gene was also positive in Western blot.Pax6 mRNA and protein expression in the Rb tumors were increased compared with normal control group (P

Interleukin-1 receptor antagonist eye drops promoting high-risk corneal allografts survival in rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Immune rejection is the main reason of grafts failure after corneal transplantation. This study was to determine whether interleukin-1 receptor antagonist (IL-1ra) eye drops could prolong corneal allografts survival in high-risk corneal orthotopic allotransplantation in rat model and to study the effect of IL-1ra on the expression of CD1-positive cells in the grafts.</p><p><b>METHODS</b>For all experiments, the Sprague-Dawley (SD) rats' corneas were transplanted into Wistar rats' eyes. High-risk transplants included those that had been sutured into Wistar recipient beds with corneal neovascularization induced by placement of three interrupted sutures in the host cornea 7 days earlier. All the animals were divided, in a masked fashion, into three treatment groups and one control group. Each treatment group received IL-1ra eye drops of different concentrations (1 mg/ml, 3 mg/ml, or 5 mg/ml, respectively) four times a day for 30 days. The control group received 0.9% normal saline (NS) eye drops in the same way as the treatment groups. All allografts were evaluated for signs of rejection from the first day after surgery. Ten days later, corneal specimens were processed to examine the expression of CD1-positive cells and histopathological changes.</p><p><b>RESULTS</b>The survival time of the transplants was 5.80 +/- 0.79, 5.89 +/- 1.05, 6.78 +/- 0.83, and 9.00 +/- 2.36 days respectively in the control or three treatment groups. Compared with the control group, 1 mg/ml IL-1ra eye drop did not prolong the survival time of the allografts (t = 0.210, P > 0.05). However, 3 mg/ml and 5 mg/ml IL-1ra eye drop did prolong the survival time of the grafts (t >or= 2.627, P < 0.05), with the latter showing more obvious effect. Immunohistochemical examinations showed a significant decrease in inflammatory cell and CD1-positive cell infiltration in IL-1ra treated groups compared with the control group.</p><p><b>CONCLUSIONS</b>IL-1ra can promote corneal allograft survival in a dose-dependant manner by reducing the infiltration of CD1-positive cells in high-risk corneal transplantation.</p>

The current status and future in limbal stem cells research / 眼科

The discovery of limbal stem cells is the key advancement in ophthalmology in the 20th century.Many clinical researches have proved that limbal stem cells transplantation is an effective method to treat severe ocular surface diseases.However, some important issues associated with the identification,isolation,standardized culture in vitro and differentiation after transplantation are still not clear and need to be paid more attention.Strengthening the underlaying research in this field will play important role in treating corneal blindness.

Study on cytokines and dendritic cells involving immunologic inhibition of rat high-risk corneal transplantation induced by SEB / 眼科

objective To explore the role of cytokines and dendritic cells (DCs) in rat high-risk corneal allograft survival pro- longed by superantigen Staphylococcal Enterotoxin B (SEB) and to compare the different effects between SEB and glucocorticoid. Design Experimental study.Participants Fisher 344 and Lewis rats.Methods The Fisher 344 and Lewis inbred rats were used for high-risk penetrating keratoplasty model.All of the Lewis rat recipients were divided into three groups by blinded fashion.GroupⅠand GroupⅡrats were injected intraperitoneally with 0.2ml saline buffer or SEB (75?g/ml) respectively at 4-day intervals on three occasions before transplantation.GroupⅢrats were injected subconjunctivally with 0.1ml dexamethasone (1mg/ml) daily from the first day after surgery for 2 weeks.The allograft survival was examined under slit-lamp.The concentration of interleukin IL-1?,IL-2,IL-4,IL-5,IL-6, IL-10,TNF-?in rat aqueous humor and peripheral blood were measured by liquichip and the cytokine and CD11c,CD80,MHC-Ⅱex- pression in corneal grafts were examined by immunohistochemestry staining.Main Octcome Measures Mean survival time of the allo- grafts,the level of cytokines in aqueous humor,peripheral blood and corneal grafts.Results Compared with group control,the grafts mean survival time was delayed about 3.8d in group SEB (P=0.00) and 7.1d in group dexamethasone(P=0.01).But there was no signifi- cant difference between group SEB and dexamethone (P=0.26).Liquichip test showed that the level of IL-1?in aqueous humor was re- duced and IL-4,IL-6 and IFN-?,ascended.Only IFN-?,and IL-6 could be found in peripheral blood,and the changing shift of them was similar to that in aqueous humor.The immunohistochemistry staining showed that the expression of IL-2 in rat corneal grafts was signif- icantly decreased but IL-4,IL-6 and IL-10 elevated.The expression of DCs in group SEB was similar to that in group control,which was elevated after keratoplasty,but the phenotype of DCs was not the same.There were predominantly mature DCs in group control while immature DCs in group SEB.Conclusions The immunological inhibiting effect of SEB is same to glucocorticoid,but the mecha- nism is different.SEB can modulate immune response,which might induce immune inhibition via the local production of cytokines and effect on DCs maturation involving corneal graft rejection prevention.