Comparison of Serum Anti-Mullerian Hormone-Level Changes in Single-Port Laparoscopic Endometriotic and Non-Endometriotic Ovarian Cyst Enucleations.

OBJECTIVES: This study compared serum anti-Mullerian hormone (AMH) levels in endometriotic cysts (ECs) with those in non-ECs and analyzed changes thereof after single-port laparoscopic (SPL) ovarian cyst enucleation using vasopressin injection. METHODS: In total, 180 patients (EC group, n = 112; non-EC group, n = 68) who underwent SPL ovarian cyst enucleation were retrospectively reviewed. Their AMH levels were checked preoperatively, on postoperative day 10 (POD10), and on postoperative month 3 (POM3). Changes in AMH levels were analyzed according to tumor type and vasopressin use. RESULTS: The median initial and postoperative serum AMH levels in the EC group were significantly lower than those in the non-EC group (preoperation: 2.0 vs 3.8 ng/mL, P < 0.001; POD10: 1.0 vs 3.2 ng/mL, P < 0.001; POM3: 1.2 vs 3.6 ng/mL, P < 0.001). The postoperative decrease in AMH levels was higher in the EC group than the non-EC group on POD10 (0.8 vs 0.5 ng/mL, P = 0.011) but not on POM3 (0.7 vs 0.5 ng/mL, P = 0.164). Vasopressin injection during EC enucleation had no significant effect on the decrease in AMH levels on POD10 (vasopressin group vs non-vasopressin group: 1.0 vs 0.8 ng/mL, P = 0.253) and POM3 (vasopressin group vs nonvasopressin group: 1.4 vs 1.1 ng/mL, P = 0.242). CONCLUSIONS: AMH levels were lower at baseline and had higher decreasing rates after SPL surgery in the EC group relative to the non-EC group. Vasopressin injection might not protect the ovary from the postoperative decrease in AMH levels.

Impact of the Learning Curve on the Survival of Abdominal or Minimally Invasive Radical Hysterectomy for Early-Stage Cervical Cancer.

PURPOSE: The objective of this study was to define the learning curve required to attain satisfactory oncologic outcomes of cervical cancer patients who were undergoing open or minimally invasive surgery for radical hysterectomy, and to analyze the correlation between the learning curve and tumor size. MATERIALS AND METHODS: Cervical cancer patients (stage IA-IIA) who underwent open radical hysterectomy (n=280) or minimal invasive radical hysterectomy (n=282) were retrospectively reviewed. The learning curve was evaluated using cumulative sum of 5-year recurrence rates. Survival outcomes were analyzed based on the operation period ("learning period," P1 vs. "skilled period," P2), operation mode, and tumor size. RESULTS: The 5-year disease-free and overall survival rates between open and minimally invasive groups were 91.8% and 89.0% (p=0.098) and 96.1% and 97.2% (p=0.944), respectively. The number of surgeries for learning period was 30 and 60 in open and minimally invasive group, respectively. P2 had better 5-year disease-free survival than P1 after adjusting for risk factors (hazard ratio, 0.392; 95% confidence interval, 0.210 to 0.734; p=0.003). All patients with tumors < 2 cm had similar 5-year disease-free survival regardless of operation mode or learning curve. Minimally invasive group presented lower survival rates than open group when tumors ≥ 2 cm in P2. Preoperative conization improved disease-free survival in patients with tumors ≥ 2 cm, especially in minimally invasive group. CONCLUSION: Minimally invasive radical hysterectomy required more cases than open group to achieve acceptable 5-year disease-free survival. When tumors ≥ 2 cm, the surgeon's proficiency affected survival outcomes in both groups.

Survival outcomes of single-port access laparoscopic radical hysterectomy for early-stage cervical cancer.

BACKGROUND: Emerging data from the Laparoscopic Approach to Cervical Cancer trial (NCT00614211) suggested that minimally invasive surgery (MIS) for cervical cancer is correlated with worse survival outcomes than open surgery. This finding could be attributed to the different learning curves for laparoscopic surgery among surgeons. This study aimed to assess the feasibility, safety, and survival outcomes of single-port access (SPA) laparoscopic radical hysterectomy (LRH) for treating early cervical cancer. METHODS: This was a retrospective cohort study of consecutive patients with early-stage cervical cancer who underwent SPA LRH between 2009 and 2018 performed by a single surgeon with expertise in SPA laparoscopy using conventional instrumentation and a homemade glove port system. RESULTS: Type C (93.2%) and B (6.8%) radical hysterectomy were performed in 59 women with cervical cancer classified as IA (3.4%), IB (94.9%), and IIA (1.7%). Forty-one patients (69.5%) had squamous cell carcinoma and 32 patients (52.5%) had tumors < 2 cm. The median operative time was 235 (125-382) minutes. There were no perioperative complications or cases of conversion to open surgery. Postoperative complications, including chylous ascites, low hemoglobin, lymphedema, and vault dehiscence, were observed in 5 patients (8.5%). Median follow-up time was 3.1 (0.6-8.6) years and 3 patients experienced recurrence (1 local and 2 distant failures). Five-year disease-free survival was 94.9% (56/59) and the 5-year overall survival rate was 98.3% (58/59). CONCLUSIONS: SPA LRH is feasible and safe for patients with early-stage cervical cancer when performed by experienced surgeons without compromising the radicality and oncologic outcomes.

Hemorrhagic bulla: a rare presentation of scabies.

Scabies is a contagious skin disorder with multiple presentations, it can also cause nosocomial infection sometimes. Bullous scabies is its rare subtype with male predilection, and typically occurs in the elderly with a median age of 70 years. We herein report a 90-year-old man who was hospitalized in the ward presenting with generalized infection of scabies associated with hemorrhagic bullae on both feet and hands, and leading to a prevalence in the ward. All the lesions including the bullae had excellent response to 10% sulfur ointment alone. No relapse occurred in more than 9 months of follow-up.

Adenosine triphosphate-based chemotherapy response assay predicts long-term survival of primary epithelial ovarian cancer.

OBJECTIVE: The aim of this study is to analyze the long-term relapse-free survival and overall survival outcomes of primary ovarian cancer patients using adenosine triphosphate-based chemotherapy response analysis. METHODS: In total, 162 primary epithelial ovarian cancer patients who underwent chemotherapy response assay for carboplatin, cisplatin, and paclitaxel by adenosine triphosphate-based chemotherapy response analysis prior to chemotherapy between December 2006 and November 2016 were retrospectively reviewed. Chemosensitivity with single or combined three agents and clinical characteristics of patients were studied to understand their correlation with long-term relapse-free survival and overall survival. RESULTS: Median follow-up time was 61.4 (range 1 - 130) months. Univariate analysis showed the paclitaxel-sensitive group (HR = 0.625, 95%CI = 0.393 to 0.994), combined carboplatin and paclitaxel-sensitive group (HR = 0.574, 95%CI = 0.352 to 0.937), and combined carboplatin, cisplatin, and paclitaxel-sensitive group (HR = 0.489, 95%CI = 0.295 to 0.810) were highly associated with better relapse-free survival than their corresponding non-sensitive groups. The carboplatin-sensitive group (HR = 0.533, 95%CI = 0.303 to 0.939), cisplatin-sensitive group (HR = 0.433. 95%CI = 0.251 to 0.748), and combined carboplatin- and cisplatin-sensitive group (HR = 0.286, 95%CI = 0.142 to 0.576) were highly associated with better overall survival than their corresponding non-sensitive groups. Combined carboplatin, cisplatin, and paclitaxel chemosensitivity, together with International Federation of Gynecology and Obstetrics (FIGO) stage were independent predictors for relapse-free survival. Single or combined chemosensitivity of cisplatin and/or carboplatin, together with residual tumor size and FIGO stage, were significant independent predictors for overall survival on a multivariate hazard model. CONCLUSION: Paclitaxel sensitivity is a prognostic factor of long-term relapse-free survival in patients with epithelial ovarian cancer, but platinum sensitivity is a more important prognostic factor for long-term overall survival.

Genetic Profiles Associated with Chemoresistance in Patient-Derived Xenograft Models of Ovarian Cancer.

PURPOSE: Recurrence and chemoresistance (CR) are the leading causes of death in patients with high-grade serous carcinoma (HGSC) of the ovary. The aim of this study was to identify genetic changes associated with CR mechanisms using a patient-derived xenograft (PDX) mouse model and genetic sequencing. MATERIALS AND METHODS: To generate a CR HGSC PDX tumor, mice bearing subcutaneously implanted HGSC PDX tumors were treated with paclitaxel and carboplatin. We compared gene expression and mutations between chemosensitive (CS) and CR PDX tumors with whole exome and RNA sequencing and selected candidate genes. Correlations between candidate gene expression and clinicopathological variables were explored using the Cancer Genome Atlas (TCGA) database and the Human Protein Atlas (THPA). RESULTS: Three CR and four CS HGSC PDX tumor models were successfully established. RNA sequencing analysis of the PDX tumors revealed that 146 genes were significantly up-regulated and 54 genes down-regulated in the CR group compared with the CS group. Whole exome sequencing analysis showed 39 mutation sites were identified which only occurred in CR group. Differential expression of SAP25, HLA-DPA1, AKT3, and PIK3R5 genes and mutation of TMEM205 and POLR2A may have important functions in the progression of ovarian cancer chemoresistance. According to TCGA data analysis, patients with high HLA-DPA1 expression were more resistant to initial chemotherapy (p=0.030; odds ratio, 1.845). CONCLUSION: We successfully established CR ovarian cancer PDX mouse models. PDX-based genetic profiling study could be used to select some candidate genes that could be targeted to overcome chemoresistance of ovarian cancer.

Functional human TSHß splice variant produced by plasma cell may be involved in the immunologic injury of thyroid in the patient with Hashimoto's thyroiditis.

Hashimoto's Thyroiditis (HT) is the most common cause of hypothyroidism in areas of the world where iodine levels are sufficient. However, the pathogenesis of HT has not been completely elucidated. The first functional human TSHß splice variant was supposed to be involved in the pathology of Hashimoto's thyroiditis. The question remains as to which kind of intrathyroid cells expresses functional TSHß splice variant and whether there are expression variations of functional TSHß splice variant in the injured thyroid of patient with HT. For the answer to this question, immune-injured thyroids were obtained from 30 patients with HT. Localization study of functional TSHß splice variant in injured thyroid was done by immunofluorescence double staining. Transcription and translation level of functional TSHß splice variant were detected by using qRT-PCR and semi-quantitative immunohistochemistry method, respectively. The correlation between expression level of functional TSHß splice variant and degree of thyroid follicles damage was assessed. It was firstly identified that functional TSHß splice variant was predominately expressed by plasma cells infiltrated around follicles and germinal center in injured thyroid of patient with HT. Of particular interest, the TSHß splice variant was expressed at significantly higher levels in the thyroid tissues of patients with HT than that in the normal thyroid tissues, furthermore, expression level of TSHß splice variant was positive related with the degree of follicles damage in thyroid of patient with HT. These findings defined the immune-derived functional TSHß splice variant that resided in the thyroid of patient with HT, which exerted the unique effects on the pathogenesis of HT, meanwhile, we considered these findings to have significant implications for understanding immune-endocrine interactions in a number of ways.

Angiotensin II receptor blocker telmisartan prevents new-onset diabetes in pre-diabetes OLETF rats on a high-fat diet: evidence of anti-diabetes action.

OBJECTIVES: This study aims to investigate the effects of telmisartan, pioglitazone and metformin administration on the prevention of new-onset type 2 diabetes mellitus in pre-diabetes Otsuka Long-Evans Tokushima Fatty (OLETF) rats fed with a high-fat diet (HFD). METHODS: OLETF rats 22 weeks of age were treated with pioglitazone (O-P), metformin (O-M), telmisartan (O-T) and low telmisartan starting from their pre-diabetes period. The weight, glucose tolerance and insulin sensitivity were measured. The lipid profiles were obtained. The abdominal subcutaneous (SC) and omental (OM) fat pads were dissected to measure the expression of mRNA and protein levels (adiponectin, proinflammatory cytokines, etc.). RESULTS: Telmisartan significantly reversed glucose tolerance and improved insulin resistance. The incidence rates of impaired glucose tolerance and type 2 diabetes in the O-P (χ(2) = 11.025, p=0.001) and O-T (χ(2)=5.495, p=0.019) groups were significantly reduced. The mRNA expression of proinflammatory cytokines was downregulated by telmisartan. The expression of adiponectin, PPARγ1 and γ2 was markedly improved by telmisartan and pioglitazone compared with the OLETF control (O-C) group. The correlation analysis showed that the systolic and diastolic blood pressures were not correlated with the homeostasis model assessment-insulin resistance (p>0.05). CONCLUSIONS: Telmisartan acts beneficially against diabetes-induced inflammation and improves insulin resistance in pre-diabetes OLETF rats fed with HFD. In view of this improved responsiveness to insulin sensitivity, telmisartan may prove to be a promising candidate for the intervention treatment of the pre-diabetes state.

Polyinosine-polycytidylic acid promotes excessive iodine intake induced thyroiditis in non-obese diabetic mice via Toll-like receptor 3 mediated inflammation.

BACKGROUND: Excessive iodine intake and viral infection are recognized as both critical factors associated with autoimmune thyroid diseases. Toll-like receptors (TLRs) have been reported to play an important role in autoimmune and inflammatory disorders. In this study, we aimed to clarify the possible mechanism of TLR3 involved in polyinosine-polycytidylic acid (poly(I:C)) promoting excessive iodine intake induced thyroiditis in non-obese diabetic (NOD) mice. METHODS: Both NOD and BALB/c mice were randomly assigned to four groups: control group (n = 5), high iodine intake (HI) group (n = 7), poly(I:C) group (n = 7) and combination of excessive iodine and poly(I:C) injection (HIP) group (n = 7). After 8 weeks, mice were weighed and blood samples were collected. All the mice were sacrificed before dissection of spleen and thyroid gland. Then, thyroid histology, thyroid secreted hormone, expression of CD3(+) cells and TLR3 as well as inflammatory mRNA level were evaluated. RESULTS: Both NOD and BALB/c mice from HI and HIP group represented goiter and increasing thyroid relative weight. Thyroid histology evidence indicated that only HIP group of NOD mice showed severe thyroiditis with lymphocytes infiltration in majority of thyroid tissue, severe damage of follicles and general fibrosis. Immunofluorescence staining results displayed a large number of CD3(+) cells in HIP NOD mice. Real-time polymerase chain reaction (PCR) results suggested interferon (IFN)-α increased over 30 folds and IFN-γ expression was doubled compared with control group, but interleukin (IL)-4 remained unchanged in HIP group of NOD mice thyroid. Meanwhile, over one third decrease of blood total thyroxine (TT4) and increased thyroid-stimulating hormone (TSH) was observed in HIP group of NOD mice. Only HIP group of NOD mice represented significantly elevation of TLR3 expression. CONCLUSION: Poly(I:C) enhanced excessive dietary iodine induced thyroiditis in NOD mice through increasing TLR3 mediated inflammation.

Tissue-engineered calcium phosphate cement in rabbit femoral condylar bone defects.

BACKGROUND: Calcium phosphate cement (CPC) is a favorable bone-graft substitute, with excellent biocompatibility and osteoconductivity. However, its reduced osteoinductive ability may limit the utility of CPC. To increase its osteoinductive potential, this study aimed to prepare tissue-engineered CPC and evaluate its use in the repair of bone defects. The fate of transplanted seed cells in vivo was observed at the same time. METHODS: Tissue-engineered CPC was prepared by seeding CPC with encapsulated bone mesenchymal stem cells (BMSCs) expressing recombinant human bone morphogenetic protein-2 (rhBMP-2) and green fluorescent protein (GFP). Tissue-engineered CPC and pure CPC were implanted into rabbit femoral condyle bone defects respectively. Twelve weeks later, radiographs, morphological observations, histomorphometrical evaluations, and in vivo tracing were performed. RESULTS: The radiographs revealed better absorption and faster new bone formation for tissue-engineered CPC than pure CPC. Morphological and histomorphometrical evaluations indicated that tissue-engineered CPC separated into numerous small blocks, with active absorption and reconstruction noted, whereas the residual CPC area was larger in the group treated with pure CPC. In the tissue-engineered CPC group, in vivo tracing revealed numerous cells expressing both GFP and rhBMP-2 that were distributed in the medullar cavity and on the surface of bony trabeculae. CONCLUSION: Tissue-engineered CPC can effectively repair bone defects, with allogenic seeded cells able to grow and differentiate in vivo after transplantation.

[Effects of naloxone on the expression of stem cell factor and C-kit receptor in combined oxygen-glucose deprivation of primary cultured human embryonic neuron in vitro].

OBJECTIVE: To explore the effects of naloxone on the expression of c-kit receptor (c-kit R) and its ligand stem cell factor (SCF) in human embryo neuronal hypoxic injury. METHODS: Serum-free cerebral cortical cultures prepared from embryonic human brains were deprived of both oxygen and glucose which would set up an environment more likely with that of in vivo ischemic injury. Neurons in 24-well culture plates were randomly divided into four groups: control group, hypoxia group, naloxone 0.5 microg/ml group and naloxone 10 microg/ml group. MTT assay and biological analysis were performed to study the cell death and the changes of extracellular concentrations of lactate dehydrogenase (LDH) after combined oxygen-glucose deprivation. Neurons in 25 ml culture flasks were also randomly allocated into four groups as previously described. Intracellular total RNA were extracted at different time points: pre-hypoxia, immediately after hypoxia, and 3, 6, 12, and 24 hours after reoxygenation. The changes of SCF/c-kit R mRNA expression in hypoxic neurons treated with different concentrations of naloxone pre and post oxygen-glucose deprivation were determined with RT-PCR. RESULTS: The cell vitality detected by MTT assay decreased significantly in hypoxia group and naloxone 0.5 microg/ml group when compared with control group (P<0.01), while no significant difference was found between naloxone 0.5 microg/ml group and hypoxia group or between naloxone 10 microg/ml group and control group. Extracellular concentration of LDH significantly increased in hypoxia group (P<0.05), while no difference was found between naloxone 0.5 microg/ml group and control group, between naloxone 0.5 microg/ml and hypoxia group, or between naloxone 10 microg/ml and control group (all P>0.05). Immediately after oxygen-glucose deprivation, the expression of SCF/c-kit R mRNA increased significantly (P<0.01). Among those the expression of SCF presented a distribution of double-peak value within 24 hours. After treated with different concentrations of naloxone, the peak value of each group were delayed to appear and went down with the increasing of naloxone concentration. The peak values in all treated groups were significantly different from that in control group (P<0.01). CONCLUSIONS: The expression of SCF/c-kit R mRNA increases at the early stage after combined oxygen-glucose deprivation. Naloxone 0.5 microg/ml can attenuate cell injuries and regulate the expression of SCF/c-kit R. Naloxone may protect neurons by modulating the expressions of some cytokines.

[Expression of truncated fragment of human OPG in CHO-DHFR- cells and its bioactivity characterization].

AIM: To obtain high level expression of recombinant human truncated osteoprotegerin (TOPG) with higher bioactivity in CHO-DHFR(-) cells. METHODS: The recombinant vector pcDNA3.1/DHFR-TOPG was constructed and transfected into CHO-DHFR(-) cells by the directions of LipofectAMINE 2000 for stable expression. The stable expression cell strains were screened by selective medium IMDM with 50 mL/L FCS, then serially passed in methotraxate (MTX) for gene amplification. The expression were analyzed by ELISA and RT-PCR. At last, the bioactivity analysis was performed in vitro. RESULTS: The expression level of recombinant truncated human OPG was up to 6 mg/L x 72 h, and it had significant suppression effect on the formation of OLC (P<0.05). CONCLUSION: Recombinant truncated human OPG has high expression and bioactivity. The results make it possible for further studying and clinical implying of OPG.

[Construction and expression of the eukaryotic expression plasmids of human TSHR extracellular domain].

AIM: To construct the eukaryotic expression plasmids of hTSHR extracellular domain and study their expression in CHO cells. METHODS: The human TSHR extracellular domain cDNAs, which were 188-403 bp and 407-904 bp, were amplified from human normal thyroid by RT-PCR. Two fragments were inserted into pcDNA3.1(D)/V5-His-TOPO.Then the recombinant plasmids pcDNA3.1-hTSHRf and pcDNA3.1-hTSHRe were transfected into CHO cells by Lipofectin after they were identified by restricting enzyme HindIII digestion analysis, PCR amplifying and DNA sequencing. RT-PCR and Western blot analysis were used to analyse hTSHR expression on mRNA and at protein levels. RESULTS: Two bands of 220 bp and 540 bp were amplified from CHO cells transfected by the recombinant plasmids pcDNA3.1-hTSHRf and pcDNA3.1-hTSHRe, respectively. Western blot analysis revealed that CHO cells transfected by pcDNA3.1-hTSHRf and pcDNA3.1-hTSHRe had strong bands with molecular weight of about 11 900 and 23 600, respectively. CONCLUSION: The recombinant plasmids have been successfully constructed. The transcription on CHO cells transfected by the recombinant plasmids has been proved by RT-PCR and eukaryotic expression has been confirmed by Western blot analysis. Our research will contribute to further study on gene expression in vivo and the establishment of animal models of Graves' disease.

[Effects of naloxone on glutamate release in combined oxygen-glucose deprivation of primary cultured human embryo neuron].

OBJECTIVE: To investigate the effects of naloxone on glutamate release in combined oxygen-glucose deprivation of primary cultured human embryo neurons. METHODS: The primary cultured embryonic human cortical neurons were demonstrated by immunocytochemical stain of neural filament (NF). The neurons were randomly allocated into control group, hypoxic group, and experimental group. The experimental group was further divided into three subgroups pretreated with different concentrations of naloxone (0.25, 5, 10 microg/ml). The neurons of hypoxic group and experimental group were deprived both oxygen and glucose for 1 hours followed by 24 hours of reoxygenation. Meanwhile, we used 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, high performance liquid chromatography (HPLC), and biological analysis to study the survival rate of neurons and the changes of extracellular glutamate and lactate dehydrogenase (LDH) levels after 24 hours of reoxygenation. RESULTS: One hour of oxygen-glucose deprivation followed by 24 hours of reoxygenation was associated with a large increase in extracellular LDH and glutamate and a significant decrease of cell vitality (P < 0.01). Naloxone exerted a concentration-dependent protection against neuronal injury provoked by combined oxygen-glucose deprivation. After reoxygenation, the extracellular concentrations of glutamate gradually decreased (P < 0.05, P < 0.01, respectively) and cell vitality increased (P < 0.01) with increase of the concentration of naloxone compared with control group. All of them returned to control level when naloxone was up to 10 microg/ml (P > 0.05). CONCLUSION: Naloxone protects neurons from hypoxic injury by inhibiting the release of glutamate and therefore alleviating the exciting toxicity.

[Anti-fungal test of composite agastache lotion on seven pathogenic fungi and its clinical application].

OBJECTIVE: To study the effect of Chinese composite recipe in treating mycotic infection. METHODS: The growth condition of 7 kinds of fungi cultured on the media containing composite agastache lotion (CAL, consisted of 5 Chinese drugs) of different concentration was observed. Result showed that CAL could inhibit 7 kinds of fungi. Based on the above anti-fungus test, 110 patients with skin tinea or genital candidiasis were treated separately with CAL, western medicine and combined (CAL and western) medicines, the therapeutic effects of the 3 groups were observed and compared. RESULTS: The therapeutic effect in patients treated with combined medicine was significantly better than that in the other two groups (P < 0.05). CONCLUSION: Combined use of CAL and western medicine could enhance the cure rate in treating skin tinea and genital candidiasis. Attention should be paid on studying Chinese anti-fungal agents.