Mechanism of Bushen Zhuyun Prescription on Improving Luteal Function of Brown Norway Rats Based on MAPKs Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the possible mechanism of Bushen Zhuyun prescription （BSZYP） to reduce the level of ovarian apoptosis in Brown Norway （BN） rats with luteal phase deficiency （LPD）. MethodFifty SPF female BN rats were randomly divided into a model group， a dydrogesterone group （0.002 g·kg-1）， and a low （4.5 g·kg-1）， a medium （9 g·kg-1）， and a high-dose （18 g·kg-1） BSZYP groups， with ten rats in each group. The rats were administrated with corresponding drugs by gavage， once a day for three estrus cycle. Western blot was used to detected the protein expression levels of c-Jun NH2 terminal kinase （JNK）， extracellular signal-regulated kinase （ERK）， phosphorylated-ERK （p-ERK）， phosphorylated-JNK （p-JNK）， p38 mitogen-activated protein kinase （p38 MAPK）， phosphorylated-p38 MAPK （p-p38 MAPK ）， B-cell lymphoma （Bcl-2）， and Bcl-2 associated X protein （Bax） in ovary. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression levels of ERK， JNK， p38 MAPK， Bax， and Bcl-2 in ovary. Enzyme-linked immunosorbent assay （ELISA） was used to detect the serum progesterone （P） and estradiol （E2） levels. Hematoxylin-eosin （HE） staining was used to observe the ovarian tissue morphology. ResultCompared with the model group， the recovery of estrus cycle of rats in all BSZYP groups had statistical significance after 1-circle administration （P<0.05）. The ovarian tissue morphology in the low-dose BSZYP group was improved， and that in the medium and high-dose BSZYP groups was significantly improved with clear follicle， less vesicular follicle and atretic follicle， and more granular layers. The number of luteum， especially the fresh luteum， in the medium and high-dose groups was increased with smooth edge and large volume. The mRNA expression level of Bcl-2 was increased in all-dose BSZYP groups， while the mRNA expression level of Bax was significantly decreased in all-dose BSZYP group （P<0.05， P<0.01）. The mRNA expression levels of JNK and p38 MAPK were significantly decreased in the high-dose BSZYP group （P<0.01）， and the mRNA expression levels of ERK were increased in the low and medium-dose BSZYP groups （P<0.05）. The protein expression level of Bcl-2 was significantly increased in the medium and high-dose BSZYP groups （P<0.01）， and the protein expression level of Bax was significantly decreased in the all-dose BSZYP groups （P<0.01）. No significant difference was observed in the protein expressions of JNK， ERK， and p38 MAPK in the BSZYP groups. The protein expression levels of p-ERK in the ovarian tissues of rats were significantly inoreased in the medium and high-dose BSZYP group （P<0.01）， p-JNK， and p-p38 MAPK in the ovarian tissues of rats were significantly decreased in the medium and high-dose BSZYP group （P<0.01）. The level of E2 was increased in all-dose BSZYP groups （P<0.05， P<0.01）， and the level of P in the medium-dose BSZYP group was increased （P<0.05）. ConclusionBSZYP improved the serum sex hormones， restored the estrous cycle， reduced atretic follicle and vesiculation， and maintained luteal morphology and function of BN rats， so as to improve luteal function and treat luteal phase deficiency. The mechanism of BSZYP may be related to reduce the level of ovarian tissue apoptosis in BN rats by regulating MAPKs signaling pathway.

Regulation of Bushen Zhuyun Prescription on Ovarian Endoplasmic Reticulum Stress in Rats with Luteal Phase Defect / 中国实验方剂学杂志

Objective:To investigate the regulatory mechanism of Bushen Zhuyun prescription（BSZYP） on endoplasmic reticulum stress （ERS） in rats with luteal phase defect （LPD） induced by mifepristone. Method:Fifty SD rats were randomly divided into a blank group， a model group， a positive control group （dydrogesterone，0.02 g·kg^-1）， and low-（0.08 g·kg^-1）and high-dose （0.24 g·kg^-1） BSZYP groups. Western blot and Real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） were used to detect the mRNA and protein expression levels of immunoglobulin binding protein （BIP）， protein kinase R-like endoplasmic reticulum kinase （PERK）， inositol-requiring enzyme 1 （IRE-1）， activating transcription factor 6 （ATF6）， and C/EBP homologous protein （CHOP）. The enzyme-linked immunosorbent assay （ELISA） was used to detect the serum progesterone （P） and estradiol （E₂） levels. Result:Compared with the blank group， the model group showed the elevated protein expression of BIP， PERK， and CHOP （<italic>P</italic><0.01） and the dwindled mRNA expression of PERK and CHOP （<italic>P</italic><0.05）， while no significant difference was observed in the protein expression of IRE-1 and ATF6， mRNA expression of IRE-1， BIP， and ATF6， and serum E₂ and P levels. Compared with the model group， the positive control group displayed diminished protein expression of CHOP （<italic>P</italic><0.01）， while no significant difference was observed in the protein expression of PERK， IRE-1， BIP， and ATF6， mRNA expression of PERK， IRE-1， BIP， ATF6， and CHOP， and serum levels of E₂ and P. The protein expression of CHOP decreased （<italic>P</italic><0.01） and the mRNA expression of CHOP increased （<italic>P</italic><0.05） in the low-dose BSZYP group， while no significant difference was observed in the mRNA and protein expression of PERK， IRE-1， BIP， and ATF6， and serum E₂ and P levels. In the high-dose BSZYP group， the protein expression of PERK， BIP， and CHOP was down-regulated （<italic>P</italic><0.01）， and the mRNA expression of CHOP was up-regulated （<italic>P</italic><0.01）， while no significant difference was observed in the protein expression of IRE-1 and ATF6， mRNA expression of PERK， IRE-1， BIP， and ATF6， and serum E₂ and P levels. Conclusion:BSZYP can treat LPD by relieving ERS.

Modified Dihuang Decoction improves ovarian reserve in mice by regulating Bcl-2-related mitochondrial apoptosis pathway / 中国中药杂志

The present study investigated the effect of Modified Dihuang Decoction in improving ovarian reserve in mice through the Bcl-2-related mitochondrial apoptosis pathway. Forty-eight adult female BALB/c mice were randomly divided into the following six groups with eight mice in each group: a blank group, a model group, a femoston group(three cycles of treatment with 0.13 mg·kg~(-1) estradiol tablets for 2 days and 1.43 mg·kg~(-1) estradiol and dydrogesterone tablets for 3 days), and high(64.74 g·kg~(-1))-, medium(43.16 g·kg~(-1))-, and low-dose(21.58 g·kg~(-1)) Modified Dihuang Decoction groups. Mice in other groups except the blank group received a single intraperitoneal injection of 12 mg·kg~(-1) cyclophosphamide and 1.2 mg·kg~(-1) busulfan to induce a model of diminished ovarian reserve(DOR), while those in the blank group received an equal volume of normal saline. Mice were treated with corresponding drugs for 15 d from the 36 th day, once per day, and the mice in the blank group and the model group were treated with an equal volume of normal saline. The general condition and oestrous cycle were observed. The serum hormone levels were detected with the enzyme-linked immunosorbent assay(ELISA). The morphological changes of ovaries were observed by HE staining. Western blot was used to detect the protein expression of cysteinyl aspartate specific proteinase-9(caspase-9), cleaved caspase-3, Bcl-2 associated X protein(Bax), Bcl-2, superoxide dismutase-2(SOD-2), and glutathione peroxidase-1(GPx-1). The mRNA expression of Bax and Bcl-2 was detected by real-time fluorescence-based quantitative polymerase chain reaction(real-time PCR). The results showed that compared with the blank group, the model group showed body weight loss, disordered oestrous cycle, elevated serum levels of follicle-stimulating hormone(FSH) and luteinizing hormone(LH), reduced serum levels of estradiol(E_2), anti-mullerian hormone(AMH), and inhibin B(INHB), the declining number of ovarian follicles and granulosa layers, increased number of atretic follicles, up-regulated protein expression of caspase-9, cleaved caspase-3, and Bax and Bax mRNA expression in ovaries, and down-regulated protein expression of Bcl-2, SOD-2 and GPx-1, and Bcl-2 mRNA expression. Compared with the model group, the Modified Dihuang Decoction groups displayed restored body weight and oestrous cycle, decreased serum levels of FSH and LH, elevated serum levels of E_2, AMH, and INHB, increased number of ovarian follicles, thickened granulosa layers, and declining number of atretic follicles. Additionally, the protein expression of caspase-9, cleaved caspase-3, and Bax, and Bax mRNA expression was down-regulated, and the protein expression of Bcl-2, SOD-2, and GPx-1, and Bcl-2 mRNA expression was up-regulated. The results suggest that Modified Dihuang Decoction can regulate endocrine hormone, promote follicle growth and improve ovarian reserve by enhancing ovarian anti-oxidant capacity, inhibiting the Bcl-2-related mitochondrial apoptosis pathway, and further inhibiting cell apoptosis.

The manifestation of videonystamography in the patients with posterior circulation ischemia vertigo / 临床耳鼻咽喉头颈外科杂志

To investigate the changes of vedionystamography(VNG)in patients with posterior circulation ischemia vertigo(PCIV).Fifty patients who complained of vertigo and imbalance with PCI were selected as experimental group for testing of visual nystamography(VNG).Thirty normal subjects were chosen as control group.The result was analyzed.The results of VNG in PCIV group and the control group were compared.The abnormal ratio were as follows:(4%,0;>0.05)for Spontaneous nystagmus,(68%,10%;<0.01)for Saccade Test,(42.0%,6.7%;<0.01)for Tracking Test,(44%,0;<0.01)for Optokinetic Test,(78%,10%;<0.01)for Positional Test,respectively.The intensity of positional nystagmus in those patients was(4.12±3.46)°/s,which was much higher than that of the control group(<0.01).One or more abnormal findings for visual-oculomotor system examination were shown in 37 patients(74%).Both vestibular central and peripheral system can be involved in PCIV.VNG test has clinical significance in differential diagnosis and lesion location.The abnormal ratio of visual nystamography in PCIV group reaches 92%(46/50).These results suggest that VNG be used as an important accessory diagnostic tool for patients with PCIV.

Regulation of BuShen ZhuYun Decotion on gonadotropin in rat pituitary cells / 医学研究生学报

Objective The mechanism of luteal phase defect remains unclear. To investigate the mechanism of BuShen ZhuYun Decoction on the gonadotropin secretion in the pituitary gland, we observed the effects of medicated serum of BuShen ZhuYun Decotion on the secretion of gonadotropin-follicle-stimulating hormone (FSH) and luteotropic hormone (LH) in rat pituitary cells.Methods The BuShen ZhuYun Decotion was administered to the female SD rats by gavage to prepare the serum containing BuShen ZhuYun Decoction. The CCK-8 method was used to detect the effect of cetrorelix acetate powder for injection, medicated serum and gonadotropin releasing hormone (GnRH) on cell activity. In the maximum non-toxic concentration, we used cetrorelix acetate powder for injection to block the GnRH receptor (GnRHR) in pituitary cells and established the GnRHR antagonistic model. Then we treat the model group with medicated serum (model group). Moreover, we established the blank group (normal pituitary cells), the cetrorelix group (intervented with cetrorelix for 6 hours), and medicated serum group (intervented with medicated serum for 24 hours). 20nmol/L GnRH was used to stimulate cells for 6h. The contents of FSH and LH in the supernatant of each group and the mRNA expression of FSHβ, LHβ and GnRHR were detected.Results Compared with that of the blank group, the supernatant levels of FSH and LH in the Cetrorelix group decreased significantly \[(3.91±0.36) mIU/mL vs (2.26±0.22) mIU/mL, (8.94±0.57) mIU/mL vs (3.35±0.59) mIU/mL, P<0.05)\]. In contrast, the levels of LH significantly increased \[(8.94±0.57) mIU/mL vs (10.79±0.60) mIU/mL, P<0.05)\]; Compared with the cetrorelix group, the levels of FSH and LH in both medicated serum group and model group increased significantly (P<0.05). Compared with the blank group, the mRNA level of FSH and LH in the cetrorelix group decreased significantly \[(0.95±0.23) mIU/mL vs (0.58±0.12) mIU/mL, (0.98±0.14) mIU/mL vs (0.27±0.21) mIU/mL, P<0.01) \], and the mRNA expression of GnRHR increased in the cetrorelix group \[(0.97±0.13) mIU/mL vs (1.77±0.26) mIU/mL, P<0.01) \]; The mRNA levels of FSH and LH in the medicated serum group were increased (P<0.05). Compared with the cetrorelix group, the mRNA expression of FSHβ mRNA and LHβ were both increased in the medicated serum group and model group (P<0.05), the mRNA expression of GnRHR decreased (P<0.01).Conclusion It is suggested that the therapeutic mechanism of BuShen ZhuYun Decotion may be related to the improvement of GnRH receptor expression.

The research progress of immunotherapy in head and neck squamous cell carcinoma / 天津医药

Despite the continuous progress and development of surgical and comprehensive treatment in recent years, the 5-year survival rate of patients with head and neck squamous cell carcinoma (HNSCC) has not been significantly improved.Therefore,improving treatment efficiency and therapeutic strategies are urgently needed.As one promising novel therapy,immunotherapy has been gradually applied in the treatment of multiple tumors including HNSCC with low toxicity and high specificity.In the immunotherapy of HNSCC,the researchers have further developed a combined systemic therapy from single cytokine therapy and achieved good effects. On the other hand, multiple cancer vaccine therapies including protein/polypeptide-dendritic cell vaccine have been put into clinical trials.In addition,immunotherapy against PD-1 and other immune checkpoints have received extensive attention,and relevant inhibitory antibodies have also been approved for the treatment of recurrent or metastatic HNSCC. Here, we briefly make a review about the progress of multiple immunotherapies for the treatment of HNSCC,including cytokine therapy,vaccine therapy and immune checkpoint therapy.

A study on the application of povidone - iodine diluent in intraoperative nursing of cervical cancer surgery / 预防医学

Objective To investigate the effect of povidone - iodine diluent on proliferation and apoptosis of cervical cancer cell HeLa and to provide the theoretical basis for its clinical application. Methods Human cervical cancer cell line HeLa in logarithmic growth phase were treated with different dilutions of povidone - iodine and the cells treated with physiological saline were set as the control group. The cells viability,morphological change,formation of apoptotic bodies,cell apoptosis and the apoptosis - related protein expression in HeLa cells were assessed by MTT assay,Hoechst33342 staining, AnnexinV / PI flow cytometry and Western blotting. Results Povidone - iodine diluent remarkably inhibited human cervical cancer cell line HeLa growth in a concentration - dependent manner. The inhibitory rates of HeLa cells were 25. 3% , 30. 8% ,33. 4% ,60. 3% ,71. 2% ,85. 3% ,89. 1% and 91. 2% when the concentration of povidone - iodine solution were 0. 001% ,0. 005% ,0. 01% ,0. 05% ,0. 1% ,0. 5% ,1% and 2% ,respectively. The nuclear chromatin of HeLa cells treated with povidone - iodine dilution was agglutinated and contracted,and the nucleus was fragile and appeared apoptotic body,with dense and dense stain or fragment dense staining. With the increase of the concentration of povidone -iodine dilution,the apoptotic rate of HeLa cells increased,so were Caspase - 8 ,Caspase - 3 and cleaved PARP. Conclusion Diluted povidone - iodine can strongly inhibit the proliferation of human cervical cancer cell line HeLa and the possible mechanism was the promotion of apoptosis.

Effects of bushen zhuyun recipe on protein expressions of estrogen receptor, progesterone receptor and integrin alpha5 and beta3 in endometrium of rats at the implantation stage / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effects of Bushen Zhuyun Recipe (BZR) on protein expressions of estrogen receptor (ER), progesterone receptor (PR) and integrin alpha5 and beta3 in endometrium of rats at the implantation stage, for exploring the possible mechanism of the recipe in treating luteal phase defect (LPD) infertility.</p><p><b>METHODS</b>Female SD rats were randomly divided into 6 groups, the blank group, the model group, the WM group treated by Western medicine, and the three BZR groups treated by low-, middle- and high-dose BZR respectively. Rats were made to pregnancy and sacrificed at the implantation stage, their middle segment of uterus, about 1 cm in length was gotten for detecting the protein expressions by Western blot. Results The protein expressions of endometrial ER and PR were significantly higher, while those of integrin alpha5 and beta3 were significantly lower than those of the control group (P < 0.05). The protein expressions of endometrial ER and PR were significantly lower, but those of integrin alpha5 and integrin beta3 were higher in rats treated by middle- and high- dose BZR than those in model rats (P < 0.05).</p><p><b>CONCLUSION</b>BZR can raise the receptivity of rats' endometrium through down-regulating the expressions of ER, PR and increasing the protein expression of integrin alpha5 and beta3 in endometrium and thus to enhance the pregnant rate.</p>

Surgical management of type II orbital neurofibromatosis / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the clinical manifestations and surgical treatment of Type II orbital neurofibromatosis.</p><p><b>METHODS</b>From Jan, 2001, to Oct, 2006, clinical data of 16 cases with type II orbital neurofibromatosis were retrospectively analyzed in the Department of Ophthalmology, Ninth People's Hospital, Medical School of Shanghai Jiao Tong University.</p><p><b>RESULTS</b>All patients had orbito-temporal or intra-orbital neurofibromatosis, combined with periorbital deformities. After partial tumor resection, the orbital reconstruction and blepharoplasty were performed in the same stage. The patients were followed up for 6-12 months with no relapse. The vision was improved in 6 cases and kept the same in 10 cases. The cosmetic results were satisfactory in all cases. 5 cases were re-operated because of relapse of blepharoptosis.</p><p><b>CONCLUSIONS</b>The function and appearance can be markedly improved after one-stage partial tumor resection, orbital reconstruction and blepharoplasty in patients with type II orbital neurofibromatosis.</p>

Effect of Chinese herb Tripterygium wilfordii Hook F monomer triptolide on apoptosis of PC12 cells induced by Abeta1-42 / 生理学报

Recent studies indicate that beta-amyloid (Abeta) is the key factor to cause neuronal degeneration in Alzheimer's disease (AD). In the present study, we set up an Abeta induced PC12 cell damage modle and studied the protective effect and related mechanisms of T(10), monomer extracted from Chinese herb Tripterygium wilfordii Hook F. PC12 cells were treated with different concentrations of Abeta (5x10(-4), 5x10(-3), 5x10(-2), 5x10(-1), 5, 50 micromol/L) for 48 h, cell viability was detected by MTT conversion. The apoptotic rate of PC12 cells was quantitatively determined using FACS assay. After PC12 cells were treated with 1x10(-11) mol/L T(10) for 48 h and then co-treated with 50 micromol/LAbetafor 48 h, the apoptotic rate and the change in intracellular Ca(2+) concentration of PC12 cells were analyzed by FACS assay and confocal, respectively. It was found that 5 micromol/L Abeta decreased the cell viability to 66.3% and 50 micromol/L Abeta decreased it to 55.1%, significantly different from that of the control group. After treatment with 50 micromol/L Abeta for 48 h, the apoptotic rate of PC12 cells increased obviously. The apoptotic rate was 5.37% in the control group, while after treatment with 0.5, 5 and 50 micromol/L Abeta for 48 h, the apoptotic rate of PC12 cells went up to 10.19%, 8.02% and 16.63%, respectively. At the same time, the concentration of intracellular Ca(2+) increased greatly after treatment with 50 micromol/L Abeta for 48 h. At the concentration of 1x10(-11) mol/L T(10) remarkably inhibited the apoptosis induced by 50 micromol/L Abeta. In the naive group, the apoptotic rate was 4.83%. The apoptotic rate went up to 17.24% after treatment with 50 micromol/L Abeta for 48 h. After co-treatment with 1x10(-11) mol/L T(10) and 50 micromol/L Abeta, the apoptotic rate decreased to 8.91%, significantly different from that of the control group. At the same time, at the concentration of 1x10(-11 )mol/L T(10) remarkably inhibited the increase of intracellular Ca(2+) concentration induced by Abeta. The results indicate that T(10) has obvious protective effect on PC12 cells, which may be related to the inhibition of the cell apoptosis and increment of intracellular Ca(2+) concentration induced by Abeta.