Objective: To establish the methods for extraction and isolation of punicalagin from pomegranate peel, and further study the purification and quantification of punicalagin. Methods: Using an ultrasonic-assisted extraction method, punicalagin in pomegranate peel was extracted at room temperature by 50% ethanol with 20-fold volume of raw material. The content of punicalagin in the crude extract was determined by HPLC. To optimize the purification process of punicalagin, static adsorption and desorption experi-ments were employed to study five kinds of macroporous adsorbent resins (D101, A8-8, NKA-9, HPD-100 and HPD-500) for the one with the highest purification efficacy of punicalagin. In addition, the technical parameters of the macroporous adsorbent resin were opti-mized to obtain punicalagin with higher purity. Punicalagin was further separated and purified by using a reverse phase MCI GEL CHP20P column. Results:HPD500 resin showed the best ability to absorb and separate punicalagin in among five kinds of macro-porous adsorbent resins. The best technical parameters were as follows:the mass concentration of sample solution was 15 mg·ml-1 , the loading amount was 2BV, the pH was 2 and the eluting solvent was 8BV of 30% ethanol. With the best process as described a-bove, the content of punicalagin extracted from pomegranate peel increased from 10. 3% to 30. 7%. The obtained punicalagin could be further purified to 61. 3% from 30% in ethanol eluate by the reverse phase MCI GEL CHP20P column. Conclusion:HPD500 resin is the most effective in the purification of punicalagin from pomegranate peel, and the content of punicalagin can be dramatically increased after the purification by a reverse phase MCI GEL CHP20P column. The optimized process shows good reproducibility and stability.

Epithelial ovarian cancer is the most lethal gynecologic malignancy, drug?resistance will eventually appear after re?peated intermittent chemotherapy. However, second?line chemotherapy only works on a few patients, new drugs with better efficacy and less side effects are needed. There are a number of promising results of molecular targeted therapy in recent clinical trials for patients with recurrent epithelial ovarian cancer, such as Bevacizumab, Olaparib, Pazopanib, etc. They have been demonstrated to improve progression?free survival of patients with platinum?resistant ovarian cancer, which might become a new strategy for such patients. In this review, we summarized the current status and progress of targeted therapy for recurrent epithelial ovarian cancer.

BACKGROUND:Transformation growth factor-β(TGF-β) and brain-derived neurotrophic factor (BDNF) are the main regulatory factors in the process of spinal cord injury. There are many researches for TGF-βand BDNF pathogenesis in the spinal cord injury, but the regulation of Ginsenoside Rg1 intervention on TGF-βand BDNF in the spinal cord injury is rarely reported.
OBJECTIVE:To observe the effect of Ginsenoside Rg1 intervention on TGF-βand BDNF expression at themolecular protein levels, and to study the protection effect of Ginsenoside Rg1 on the spinal cord and nerve function after spinal cord injury.
METHODS:Experimental rats were randomly divided into blank control group, model group and Ginsenoside Rg1 group. In the model and Ginsenoside Rg1 groups, spinal cord injury model was established with the impact method in rats. In the Ginsenoside Rg1 group, rats were intraperitoneal y injected with 10 mg/kg Ginsenoside Rg1 24 hours after modeling, once per day, for 14 days. Rats in the blank control and model groups were injected with equal saline.
RESULTS AND CONCLUSION:Compared with the control group, serum malondialdehyde levels increased, the content of superoxide dismutase decreased, TGF-βexpression levels in spinal cord tissue increased, and BDNF expression levels decreased in the model and Ginsenoside Rg1 groups. Compared with the model group, serum malondialdehyde levels decreased, the content of superoxide dismutase increased, TGF-βexpression levels in spinal cord tissue decreased, and BDNF expression levels increased in the Ginsenoside Rg1 group. Ginsenoside Rg1 can protect the injury spinal cord in rats after spinal cord injury.

BACKGROUND:Chinese herb extracts can restore and protect the nervous system of rats through intervention of neural stem cels. OBJECTIVE:To explore the role of ginsenosides Rg1 in the proliferation and protection of neural stem cels. METHOD:Sprague-Dawley rats at pregnant 19 days were dissected to take out fetal rats, and then the hippocampal tissues from fetal rats were isolated to extract neural stem cels. Neural stem cels were co-cultured with DMEM/F12 medium containing 50 g/L ginsenosides Rg1 as intervention group, with DMEM/F12 medium as blank control group, and with DMEM/F12 containing 0.64% phenol as positive control group, respectively. MTT assay was used to detect the proliferation of neural stem cels in each group, and western blot method to detect the protein expression of brain-derived neurotrophic factor and transforming growth factor-β in neural stem cels. RESULTS AND CONCLUSION:Rat neural stem cels were round single cels with clear border at early period after isolation but at 2 days after inoculation, the cels were adherent and aggregated into smal cel spheres. Compared with the blank control group, the proliferative rate of neural stem cels was significantly increased in the ginsenosides Rg1 group (P < 0.05), but decreased in the positive control group (P < 0.05). Compared with the blank control group, in the ginsenosides Rg1 group, the expression of brain-derived neurotrophic factor was elevated, and the expression of transforming growth factor-β was reduced, indicating ginsenosides Rg1 has a certain effect to promote the proliferation of neural stem cels as wel as to protect the neural stem cels.

OBJECTIVETo investigate the synergistic effect of transforming growth factor-beta1 (TGF-beta1) and platelet-derived growth factor-BB (PDGF-BB) on the expression of integrin beta3, in periodontal membrane of rat orthodontic tooth.

METHODSAn orthodontic tooth movement model was established. Up to 32 experimental rats were randomly divided into four groups according to a random number table. The four groups were injected with 1% PBS, TGF-beta1 (5 ng), PDGF-BB (10 ng), and combined TGF-beta1 (5 ng) and PDGF-BB (10 ng) in the buccal submucosal, respectively. The volume injected in each group was 0.1 mL. The animals were then sacrificed on the 10th day. The left maxillary first molar and periodontal tissue were taken. Different expressions of integrin beta3 were detected in periodontal tissues through immunohistochemistry. Mean optical density (OD) values of the positive fields were examined. The data obtained were analyzed through ANOVA. The data followed normal distribution, and were compared via t-test.

RESULTSCompared with the control groups, the expression of integrin beta3 was higher in the experimental groupin tension sides (P < 0.01). Significant differences in tension sides between the single-injection groups and the combined group were observed (P < 0.01). Compared with the control groups, the expression of integrin beta3 was higher in the experimental group in compression sides (P < 0.05). In addition, there was no significant differences in compression sides between the single-injection groups and the combined group (P > 0.05).

CONCLUSIONIn terms of local regulatory factors, TGF-beta1 combined with PDGF-BB enhance the expression of integrin beta3 in the periodontal membrane and accelerate periodontal remodeling. The synergistic effect of the two growth factors is better than the single growth factor.

Objective To explore the effects of human umbilical cord mesenchymal stem cells (HUC?MSCs) on podocytic apoptosis and injury induced by high glucose (HG) and the underlying mechanisms. Methods Podocytes were divided into six groups according to treatment: ⑴ normal glucose group (NG);⑵high glucose group (HG);⑶mannitol control group (NG+Ma);⑷HUC?MSC co?culture group (HUC?MSCs); ⑸ recombinant human hepatocyte growth factor treatment group (rhHGF);⑹ neutralizing antibody group(HGF?NtAb). Cytometry and Hoechst staining were used to detect the apoptosis rates. Western blot was used to measure the ratio of active PARP to total PARP and the level of Bcl?2. Immunofluorescence was used to study podocytic apoptosis and injury. Neutralizing antibody (NtAb) was used to block its function and the recombinant cytokine was added to induce its function. Results High glucose induced podocytic apoptosis in a time?dependent manner, HUC?MSCs co?culture decreased the podocytic apoptosis rate and the expression of PARP (all P﹤0.05), increased the expression of Bcl?2, prevented the reduced expression and maintained the normal arrangement of podocytic podoplanin. The rhHGF prevented podocytic apoptosis and injury similarly to HUC?MSCs, the beneficial effect of HUC?MSC decreased when blockade of HGF. Conclusions HUC?MSCs co?culture ameliorates podocytic apoptosis and injure induced by HG, probably through secreting soluble HGF.

Purpose To investigate the clinicopathlogical characteristics, diagnosis and differential diagnosis of ovarian small cell car-cinoma hypercalcemic type ( OSCCHT) associated with acute renal dysfunction. Methods A case of OSCCHT associated with acute renal dysfunction was reported. The clinical and pathologic data, treatment and pathological examinations were analyzed and the related literatures were reviewed. Results A 29-year-olds women was presented to hospital with inappetence and significant weight loss for 2 months. The laboratory examination showed abnormal renal function, and pelvic cavity mass, possibly coming from adnexa of the uterus was seen by radiography. Serum levels showed significantly increased serum calcium with acute renal dysfunction. An emergency opera-tion was performed just after renal function partially recovered by 2 times hemodialysis. During surgery, right ovary tumor with a size of 12 cm × 10 cm × 10 cm was inspected. Microscopically, the tumor cells were arranged in a diffuse solid pattern, mutiple nodules were separated by fibrous tissue. some small folliculars and pseudoglandular cavities with acidophilia secretion within nodules could been ob-served. Tumor cells were medium to large with eosinophilic cytoplasm, round or oval vesicular nucleus, increased karyoplasmic ratio and pathologic mitosis. Immunohistochemistry revealed that the tumor cells expressed EMA, CKpan, C-erbB-2 ( +) , CA125 ( focal+) and Ki-67 proliferation index was about 60%, while ER, PR, Syn, CgA, PTH, Inhibin, CD99, AFP, PLAP, CD30 and CD20 were not expressed. Conclusion Ovarian small cell carcinoma hypercalcemic type with acute renal dysfunction is a very rare with a rapidly progressive and highly malignant tumor. The final diagnosis mainly lies on clinical information ( hypercalcaemia) , morphology and immunohistochemistry, combined with electron microscopy and molecular biological detection when necessary.

Objective Although the correlation between high risk human papilloma virus (hrHPV) infection and cervical cancer ( CC ) or cervical intraepithelial neoplasia ( CIN ) is well known , vaginal cancer ( VC ) or vaginal intraepithelial neoplasia ( VAIN) also caused by hrHPV has not received enough attention .This article aims to explore the clinical characteristics of VC or VAIN after operations of CC or CIN in order to provide evidence for the treatment of these diseases . Methods The clinical charac-teristics and treatment of 15 cases with VC or VAIN after operations of CC or CIN were reviewed from Jan 2010 to May 2013 in our hos-pital. Results The mean age was (53.6 ±10.82) years, ranged from 39 to 73 years.The duration from the first operation to devel-oped VAIN or VC was (25.07 ±18.31) months, ranged from 1 to 60 months.There are 4 cases developed VC, 4 cases VAINⅢand 2 cases VINⅡfrom 10 CC patients;and 3 cases developed VC , 2 cases VAINⅢfrom 5 CINⅢpatients.hrHPV test were positive in all 15 patients.Treatment in these series were performed including total vaginectomy in 8 patients (3 VC, 4 VAINⅢ and 1 VAINⅡpatients), pelvic lymphonectomy in 1;upper vaginectomy in 2 patients (1 VC, 1 VAINⅢ), radiation or chemo-radiation therapy in 3 (3 VC), interferon muscle injection combined with topical application of estrogen and acyclovir gel in 2 (1 VC, 1 VAINⅡ). Conclusion Careful follow-up after CC or CIN operations are very important because continued hrHPV infection may result VC and VAIN lesions.Vaginectomy may be the best therapy .Interferon muscle injection combined with topical application of estrogen and acyclovir gel are also alternatively therapy , especially for hard to operate patients . Radiation therapy seems to be not very adaptable for VAIN patients .

OBJECTIVE: To explore the effects of Salvia miltiorrhiza on renal morphology and renal function of rats with streptozotocin diabetes. METHODS: Thirty male Wistar rats were randomly divided into three groups, which were normal control group, untreated group and Salvia miltiorrhiza-treated group. Diabetic nephropathy was induced in rats of the last two groups by intraperitoneal injection of streptozotocin after unilateral nephrectomy. Then the rats in the normal control and untreated groups were fed with normal saline while those in the Salvia miltiorrhiza-treated group were fed Salvia miltiorrhiza preparation for 8 weeks. The glomerular volume (VG), kidney-to-body weight ratio (KW/BW), urinary albumin excretion rate (UAER) and creatinine clearance (Ccr) were observed. The expression levels of transforming growth factor-beta1 (TGF-beta1), connective tissue growth factor (CTGF), fibronectin (FN) and plasminogen activator inhibitor-1 (PAI-1) were detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) at the end of the experiment. RESULTS: UAER, Ccr, VG and KW/BW ratio were significantly higher in the untreated group than those in the normal control group (P<0.05). The expression levels of TGF-beta1, CTGF, PAI-1 and FN in the untreated group were also significantly higher as compared with those in the normal control group (P<0.05). UAER, Ccr, VG, KW/BW ratio and the levels of TGF-beta1, CTGF, PAI-1 and FN in the Salvia miltiorrhiza-treated group were obviously lower than those in the untreated group (P<0.05). CONCLUSION: Salvia miltiorrhiza can protect rats with streptozotocin diabetes from diabetic nephropathy by suppressing the over-expressions of TGF-beta1, CTGF, PAI-1 and FN in renal cortex.

OBJECTlVE To estabIish a simpIe,sensitive and quick method for determination of B7011 in rat pIasma. METHODS The method of protein precipitation with methanoI was used for pre-treatment of pIasma sampIes determined by Iiquid chromatography mass spectrometer. The Iinear reIa-tionship,intra-batch and inter-batch precision,specificity,matrix effect,recovery rate,the accuracy and stabiIity of the pIasma sampIes were vaIidated. The concentration of B7011 in pIasma was determined by LC-mS/ mS foIIowing a singIe intravenous injection of B7011 0.5 mg·kg-1 to rats. RESULTS The Iinear range of B7011 was 30-20 000 μg·L-1 ,the Iower Iimit of quantification was 30 μg·L-1 in pIasma,the in-tra-batch precision of 60,1000,16 000 and 10 000 ng·mL-1 was 5.61% -13.31%,2.31% -8.35%, 2.02%-9.47% and 4.0%-15.0% respectiveIy,and inter-batch precision was 10.05%,2.55%,3.75% and 8.58% respectiveIy. The recovery of 60,1000,and 16 000 μg·L-1 was 114.12%,109.2% and 101.06%respectiveIy. The average peak concentrations were 8373.28 and 8564.59 μg·L-1 ,the mean AUC was 98 400 and 104 000 μg·L-1·h and the t1/ 2z for B7011 was 41.7 and 63.6 h in bIood of maIe and femaIe rats, respectiveIy. CONCLUSlON The estabIished method is sensitive, fast and simpIe and concentration of B7011 in pIasma is determined by LC-mS/ mS foIIowing a singIe intravenous injection of B7011 0.5 mg·kg-1 to rats. It can satisfy the requirements of pharmacokinetic and toxicokinetic studies.