Objective To investigate the relationship between cysteine-rich protein 61 ( Cyr61 ) and phosphatidylinositol 3-kinase ( PI3K ) signal pathway on cell proliferation and apoptotic in human ovarian carcinoma cells.Methods Recombinant human Cyr61 (rhCyr61) was pretreated with ovarian carcinoma cells.The expression of Cyr61 protein was detected by confocal spectral microscopy.Then treated the ovarian carcinoma cells with PI3K transduction inhibitors (LY294002) for 24 hours.Cell apoptosis was detected by flow cytometry (FCM).Cell viability was determined by methyl thiazolyl tetrazolium (MTT) method.The mRNA expressions of Cyr61,the protein levels of protein kinase B ( PKB),phospho-PKB and Cyr61 were assaved by real time-PCR and western blot analysis,respectively.Results The Cyr61 and phospho-PKB protein expression in two ovarian carcinoma cells (OV2008 and OVCAR-3 ) were increased in rhCyr61pretreated group.The decreasing of cell apoptosis [ ( 1.4 ±0.9)％,(2.1 ± 1.0)％ ] and increasing of cell proliferation [ ( 124.0 ± 1.8)％,( 133.0 ±2.2)％ ] was detected in the same time,compared with negative control group,there were significant difference ( P ＜ 0.05 ).After exposed to LY294002 for 24 hours,the apoptosis rate of OV2008 and OVCAR-3 in pretreated with rhCyr61 group exposed to LY294002 were (21.1 ± 1.6)％ and (26.4 ± 1.5 )％,respectively.Cells viability [ (59.0 ± 2.3 )％,(51.0 ± 2.0)％ ]was also significantly decreased in OV2008 and OVCAR-3 pretreated with rhCyr61 cells.Meanwhile,the mRNA expressions of Cyr61 (3.2 ± 0.8,6.2 ± 1.1 ) and the protein levels of phospho-PKB and Cyr61 were greatly decreased.Compared with negative control group,there were significant difference in OV2008 and OVCAR-3 cells (all P ＜ 0.0l ).Conclusions The activation of PI3K intracellular signaling pathways may lead to up-regulation of Cyr61 expression.Block PI3K signal pathway could significantly inhibit the expression of Cyr61,and may promote the apoptotic effects and inhibit the cell growth of ovarian carcinoma cells.

Objective To investigate the expression of steroidogenic factor 1 ( SF-1 ) in eutopic endometrium,ovarian endometriosis (EM) and adenomyosis.Methods From Jan.2008 to Dec.2010,30 patients with both ovarian endometriotic cysts and adenomyosis underwent hysterectomy,cystectomy or adenxectomy in Peking University First Hospital.The combination of ovarian EM and denomyosis were confirmed in 17 cases.In the mean time,endometria form 10 patients with cervical intraepithelial neoplasm ( CIN ) Ⅲ underwent hysterectomy were selected as controls.The expression of SF-l in eutopic endometrium,ovarian EM and adenomyosis was detected by immunohistochemistry staining.Results No immunoreactivity of SF-l was observed glandular cell and stromal cell in eutopic endometrium of both groups.14/17 of positive rate of SF-l were observed in lesions of ovarian endometriosis.SF-l protein was not expressed in the endometrial glandular cells of ovarian EM and in adenomyosis foci.Conclusion The differences of SF-l expression in ovarian endometrioma and adenomyosis foci might mediate the development of the disease.

Erchen Decoction is widely used to dampness-phlegm syndrome and it can be flexibly used in clinical application by my supervisor Professor OU Zheng-wu, especially in cough, pneumonia, asthma and influenza, which has obvious efficacy in treatment. This article reviewed five medical cases during the follow-up process with Professor OU in application of Erchen Decoction, with a purpose to share experience of famous doctors and improve TCM diagnosis and treatment level together.

Epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) targeted treatment has been a standard therapy for advanced non-small cell lung cancer (NSCLC), but it is not tolerated well by all patients. In China, some studies have reported that traditional Chinese medicinal herbs (TCMHs) may increase efficacy and reduce toxicity when combined with EGFR-TKI, but outside of China few studies of this kind have been attempted.

BACKGROUND:Rapid loss of liver-specific functions of the cultured hepatocytes limits the development of hepatocyte-based therapies. OBJECTIVE:To establish a three-dimensional culture system based on col agen hydrogel that enables to enhance liver-specific functions for a long period during culture of hepatocytes. METHODS:Hepatocytes were isolated from Sprague-Dawley rats and then encapsulated into liquid type Ⅰcol agen solution that was premixed with hepatocyte growth factor and Dulbecco's modified Eagle’s medium to create hepatocyte/col agen hydrogel compounds. The compounds were inoculated into a 96-wel plate. After gelation, culture medium was added. Light microscope, hematoxylin-eosin staining and transmission electron microscopy were used to examine the morphological characteristics and ultrastructure of the cultured hepatocytes. The cellsupernatant was col ected and tested for albumin secretion and urea synthesis. Periodic acid-schiff staining, immunofluorescence staining and quantitative real-time PCR were also used to further clarify liver-specific phenotype or function of the hepatocytes.RESULTS AND CONCLUSION:(1) Light microscope revealed that hepatocytes were round shape and distributed uniformly in col agen hydrogel. The three-dimensional hepatocyte culture system exhibited similarities to liver-like structure and tight junction were formed between hepatocytes after 14 days of culture. (2) Within the three-dimensional culture system, hepatocytes remained positive for periodic acid-schiff staining, albumin and hepatocyte nuclear factor-4αpositive after 14-day culture, which provided the convincing evidence of highly differentiated primary hepatocytes with functions of glycogen and albumin synthesis. (3) The albumin and urea productions in the three-dimensional culture system had a significantly higher level than in the two-dimensional culture, and could remain at a high level at least for 15 days. (4) The expression levels of hepatocyte-specific genes including Albumin, HNF-4α, Claudin-3, CYP1A1, CYP3A1 and G6P were significantly improved in the three-dimensional culture as compared with the two-dimensional culture. The col agen hydrogel based three-dimensional culture system provides a valuable model to enhance the hepatocyte functional maintenance and lay the foundation for the development of hepatocyte-based therapy for liver disease.

OBJECTIVETo analyze the rules for acupoint selection of acupuncture and moxibustion in domestic clinical treatment of perimenopausal syndrome based on data mining technology in modern times.

METHODSThe relevant literature were retrieved from Chinese Biomedical Literature Database (CBM), China National Knowledge Infrastructure (CNKI) and Wanfang database on this disease treated with clinical acupuncture and moxibustion in China from 1978 to 2013. The database of acupuncture-moxibustion prescription was set up. The relevant regulations of data mining technology were used to analyze the rules for acupoint selection.

RESULTSTotally, 211 papers, 254 acupuncture-moxibustion prescriptions and 130 acupoints were included. The total frequency of acupoints application was 2193 times, with 14 meridians involved. The utilization of the acupoints in the lower limbs and on the back were 33.0% (723/2193) and 23.8% (521/2193) and those of yin and yang meridians were 51.8% (1136/2193) and 44.0% (965/2193), respectively. The utilization of the specific acupoints accounted for 88.7% (1946/2193).

CONCLUSIONIn clinical treatment of perimenopausal syndrome with acupuncture and moxibustion in modern times, the acupoint selection from involved meridians is the basis, associated with multiple methods of acupoint combination; yin and yang meridians are equally important and the specific acupoints are considered particularly critical in application.

Acupuncture Points ; Acupuncture Therapy ; China ; Clinical Trials as Topic ; Female ; Humans ; Perimenopause ; physiology

OBJECTIVETo explore the role of Compound Danshen Injection (CDI) in regulating the expression of aquaporin 3 (AQP3) in human amnion epithelium cells (hAECs), and to study the relation between c-Jun N-terminal kinase (JNK) signal pathway and AQP3.

METHODShAECs were isolated and primarily cultured from term pregnancy with normal amniotic fluid volume and from term pregnancy with oligohydramnios, and then hAECs were further divided into four groups, i.e., the blank control group (A), the SP600125 group (B), the CDI group (C), and the SP600125 +CDI group (D). The cell viability was measured by cell counting kit-8 assay (CCK-8). The expression of total JNK, phosphorylated JNK, and AQP3 were determined by Western blot.

RESULTS(1) In hAECs with normal AFV or with oligohydramnios: There was no statistical difference in the cell viability or the expression of total JNK among the 4 groups (P > 0.05). But there was statistical difference in the expression of p-JNK (P < 0.05). Compared with A group, the expression of p-JNK was obviously down-regulated in B group, but obviously up-regulated in C group (P < 0.05). The expression of p-JNK was significantly lower in D group than in C group, but higher than that in A group or B group (P < 0.05).The AQP3 expression in the hAECs with normal amniotic fluid volume of C group and D group were higher than that in the A group (P < 0.05). However, there was no statistical difference in the AQP3 expression between C group and D group (P > 0.05). In hAECs with oligohydramnios, the expression of AQP3 obviously decreased in B group, but up-regulated in C group (both P < 0.05). The expression of AQP3 was lower in D group than in C group, but higher than in B group (P < 0.05).

CONCLUSIONCDI could regulate the AQP3 expression in hAECs with oligohydramnios via activating the JNK signal pathway.

Amnion ; cytology ; drug effects ; Aquaporin 3 ; metabolism ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; drug effects ; metabolism ; Female ; Humans ; JNK Mitogen-Activated Protein Kinases ; metabolism ; MAP Kinase Signaling System ; physiology

Objective To explore the value of transrectal ultrasound (TRS) in diagnosing middle ureteral calculi.Methods Patients who were suspected as middle urinary calculi by clinicians underwent transabdominal ultrasound (TAS) and transrectal ultrasound in the First Affiliated Hospital of Guangzhou Medical University and Guangzhou Hospital of Integrated Traditional and West Medicine from 2011 to 2012. The diagnosis were finally confirmed by a variety of methods .Results Forty patients were eventually diagnosed as middle ureteral calculi by X-ray,computer tomography (CT),surgery,intravenous pyelography (IVP)imaging and magnetic resonance imaging (MRI).The other 5 cases were diagnosed as ureteral tumor , ureteral cystis(confirmed by MRI)and oppressed ureteral by gynecology cystis (confirmed by surgery).TRS detected 37 of 40 middle ureteral calculi(92.5%).TAS detected 8 of 40 middle ureteral calculi(20.0%). The detection ratio of TRS was higher than that of TAS and the difference was statistically significant (P=0.00).TAS could display ureter across iliac blood vessels expansion in all 40 patients.Conclusions Using TRS in diagnosing middle ureteric calculi is quick , convenient with high sensitivity and worthy of wide application.TAS can display ureter across iliac blood vessels expansion and can help to diagnosis of middle ureteral calculi.

Objective To investigate the clinical states of enuresis children companied with spina bifida occulta(SBO)and study the efficient way of treatment.Methods The children with SBO were check out by X ray from a total of 121 children with bedwetting.Their parents were asked to complete the enuresis questionnaires.Urine routine test and B-ultrasound examination about kidney,bladder and ureter were also asked to be done.The clinic data of the 49 children were attained and analyzed.They were randomly divided into 2 groups,and given the controlled treatment.Group A[used 1-deamino-8-D-arginine vas-opressin(DDAVP)only] and group B(used DDAVP plus oxybutynin plus bladder training)treated for 12 weeks.Results There were totally 49 bedwetting children companied with SBO,and most of them(44 cases,89.8%)were severe type(bedwetting times≥7 times/week).Some of them coexisted with frequency,urgency,gentle urgency incontinence and microscopic hematuria(22 cases).Thirty cases were found the functional bladder capacity(FBC)decrease by B-ultrasound.The cure rates were 58.3%(group A)and 88.0%(group B)respectively.The relapse rates were 36.8%(group A)and 12.5%(group B)respectively after stopping treatment for 3 months.Conclusions SBO accounts for considerably higher rate in enuretic children.It might cause the disability of bladder function.The treatment plan with DDAVP plus oxybutynin plus bladder training can not only increase the cure rate but also lower the relapse rate.

italic>Cynanchum wallichii and Cynanchum otophyllum belong to the genus Cynanchum in the family Apocynaceae, and are important medicinal plants. In this study, we sequenced and assembled the chloroplast genomes of C. wallichii and C. otophyllum, and performed a phylogenetic analysis of the structural characteristics of their chloroplast genomes and their phylogenetic positions. The results showed that the chloroplast genomes of both C. wallichii and C. otophyllum had a typical tetrad structure, with 133 genes annotated, and the total GC contents of both were similar. Codon preference analysis showed that the relative synonymous codon usage in the chloroplast genomes of C. wallichii and C. otophyllum differed slightly, but the differences were not significant, and there was a strong A or U preference at the third codon position. In both chloroplast genomes, 91 and 103 simple sequence repeats were detected respectively, and the largest proportion of A/T type repeats. Nucleotide polymorphism analysis showed that the nucleotide diversity of the intergenic sequences in the chloroplast genome of genus Cynanchum were generally higher than those of the common gene sequences. A pair of primers was designed based on the high variation region of the chloroplast genome to identify C. wallichii and C. otophyllum. The phylogenetic analysis showed that the C. wallichii and Cynanchum thesioides were the closest relatives, while the C. otophyllum, Cynanchum bungei and Cynanchum wilfordii formed a stable monophyletic clade within the genus Cynanchum, and the three species were closely related. The comparative analysis of the chloroplast genomic characteristics and phylogeny of C. wallichii and C. otophyllum will provide a theoretical basis for the species identification of the two plants and for the study of genetic diversity and phylogeny of the genus Cynanchum.