Clomiphene citrate mild stimulation improved follicular development outcomes in PCOS women with high luteinizing hormone and poor ovarian response: A case report.

INTRODUCTION: Polycystic ovary syndrome (PCOS) is a main cause of anovulatory infertility in women of reproductive age. About 30% to 50% of patients with PCOS has high serum basal luteinizing hormone (LH) levels, and almost 5% of PCOS women with high LH have poor ovarian response (POR). We reported a case of a PCOS woman with high basal LH levels who canceled due to POR during two consecutive controlled ovarian stimulation treatments, which was considered to be related to the suppression of LH levels during downregulation. Clomiphene citrate (CC) combined with human menopausal urinary gonadotropin (HMG) mild regimen did not affect LH levels and obtained good follicular development, providing a new treatment insight for patients with PCOS combined with POR. PATIENT CONCERNS: A 28-year-old PCOS woman with high basal LH levels, underwent IVF assisted pregnancy treatment in our hospital, whom canceled due to POR during two traditional controlled ovulation induction program. Follicular development was finally achieved with CC milder protocol. DIAGNOSIS: This patient with the diagnosis of PCOS was undergone IVF assisted pregnancy treatment in our hospital. INTERVENTIONS: CC protocol supports the development of follicular. OUTCOMES: CC protocol resulted in better follicular development and high-quality embryos due to the continuous maintenance of an elevated LH levels. CONCLUSION: PCOS women with poor ovarian response required relatively higher LH to maintain the normal development of follicles.

Pregnancy confirmed after controlled ovarian stimulation for infertility: A case report.

RATIONAL: Induction of ovarian stimulation by use of the gonadotropin-releasing hormone agonist (GnRHa) long protocol in the luteal phase is a common practice and results in stable pregnancy and live births; it is often used in patients with normal ovarian function. Some patients with normal ovulation may be pregnant before ovulation induction, which can be easily confirmed by asking the patient about cessation of menstruation. However, some pregnancy complications may cause vaginal bleeding along with normal menstrual blood loss; in such a situation, hormone levels can often mirror that seen in pituitary down-regulation and the value of ß-HCG may be less than 5 mIU/mL. Under these conditions, the physician might start the cycle of ovarian stimulation. During ovarian stimulation, the increase in ß-HCG can cause premature luteinization and follicle maturation disorder, and poor embryo quality, which can easily be overlooked. In this study, we report a case of pregnancy at the end of controlled ovarian stimulation induced by GnRHa long protocol in the luteal phase, followed by follicle maturation disorder and poor embryo quality. This case provided a reference and served as a cautionary note that could perhaps obviate occurrence of similar cases. PATIENT CONCERNS: A 30-year-old woman with a diagnosis of unexplained infertility was scheduled for in vitro fertilization embryo culture (IVF) at our clinic. Pregnancy was confirmed at the end of controlled ovarian stimulation, which was followed by follicular maturation disorder and poor embryo quality. DIAGNOSIS: The patient with a diagnosis of unexplained infertility was scheduled for IVF at our clinic. INTERVENTIONS: Oocyte retrieval was still arranged for her after confirmation of pregnancy. As per the ß-HCG level and the trans-vaginal ultrasound examination findings, we considered 2 possibilities: an adverse intrauterine pregnancy or extra-uterine pregnancy. Therefore, we decided to terminate the pregnancy; hence, 50 mg/d of mifepristone was given for 2 days, combined with 200 µg misoprostol. OUTCOMES: Elevated ß-HCG level had an adverse effect on maturation and fertilization of oocytes, and even embryo quality. CONCLUSION: Once pregnancy is confirmed, ovulation induction should be terminated as soon as possible.

Anti-Müllerian Hormone Accelerates Pathological Process of Insulin Resistance in Polycystic Ovary Syndrome Patients.

Insulin resistance (IR) is one of the most common features of polycystic ovary syndrome (PCOS), which is related to obesity. Whether increased anti-Müllerian hormone (AMH) levels in PCOS are involved in the pathogenesis of insulin resistance remains unclear. We investigated serum levels of leptin and AMH along with basic clinical and metabolic parameters in 114 PCOS patients and 181 non-PCOS women. PCOS patients presented higher fasting blood glucose, insulin concentrations and Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) in addition to body mass index (BMI), lipids profiles and hormone levels. HOMA-IR showed a positive correlation with BMI, AMH, leptin, and low-density lipoprotein-cholesterol (LDL-c) levels. Interestingly, AMH is strongly positively correlated with HOMA-IR and insulin concentrations for 1st and 2nd hours of glucose treatment after fasting. Among PCOS women with BMI≥25 kg/m2, high AMH level group showed an increased HOMA-IR when compared to normal AMH level. However, among PCOS women with normal BMI, women with high AMH presented an elevated fasting insulin levels but not HOMA-IR when compared to normal AMH group. In vitro treatment of isolated islet cells with high concentration of leptin (200 ng/ml) or high leptin plus high concentration of AMH (1 ng/ml) significantly enhanced insulin secretion. Importantly, co-treatment of AMH plus leptin upregulates the expression of pro-apoptotic proteins, such as Bax, caspase-3, and caspase-8 after incubating with a high level of glucose. These results suggest that AMH may involve in the pathological process of pancreatic ß-cells in obese PCOS women.

Onset of HLA-B27-associated diseases in diabetic patient during a period of religious fasting: A case report.

RATIONALE: The association between human leukocyte antigen B27 (HLA-B27) with its associated diseases is far from complete. The role of HLA-B27 in disease susceptibility is still not known, although many suggestions have been proposed. PATIENT CONCERNS: The patient was a 46-year-old policeman with a history of obesity, diabetes, and hypertension. He was a Shaolin lay disciple who fasted at the Shaolin temple for at least 1 week each year since 2014. DIAGNOSES: The patient suffered three different HLA-B27-associated diseases including acute anterior uveitis, ulcerative colitis, and ankylosing spondylitis, from 2014 to 2016 because of prolonged fasting. INTERVENTIONS: The patient accept standard treatment after the diagnosis of acute anterior uveitis, ulcerative colitis, and ankylosing spondylitis. OUTCOMES: The patient's symptoms and signs of acute anterior uveitis, ulcerative colitis, and ankylosing spondylitis were all relieved within one week after the clinical treatment. LESSONS: Our case suggested that prolonged fasting may lead to the onset of HLA-B27-associated diseases in diabetic patient.

Neutrophil extracellular traps involvement in corneal fungal infection.

PURPOSE: Neutrophils release neutrophil extracellular traps (NETs) when defending against invading microorganisms. We investigated the existence of NETs in fungal keratitis. METHODS: Fourteen patients with unilateral fungal keratitis were included. Detailed information about each patient was recorded, including (1) patient history (onset of symptoms and previous therapy), (2) ocular examination findings by slit-lamp biomicroscopy, (3) laboratory findings from direct smear examination and culture of corneal scrapings, (4) NET formation, and (5) treatment strategy and prognosis. Immunofluorescence staining was used to evaluate the existence of NETs on corneal scrapings. The relationship between the quantification of NETs and the clinical character of the fungal keratitis was identified. RESULTS: NETs were identified in all 14 patients. Patients with a higher grade of NET formation and fewer fungal hyphae always showed a good treatment response and a short course of infection. NETs were consistently found mixed with fungal hyphae in the corneal scrapings from infected patients. No statistical significance was found between the grade of NETs formed and the course of infection before presentation, and no relationship between the quantification of NETs and the size of the ulcer was found. CONCLUSIONS: The results suggest that NETs are involved in fungal keratitis. The number of NETs in infected corneas may provide a tool for evaluating the prognosis for fungal keratitis.

Hormone replacement therapy benefits meibomian gland dysfunction in perimenopausal women.

Meibomian gland dysfunction (MGD) is believed to be the leading cause of dry eye (DE) disease worldwide. The connection between aging and MGD has long been recognized. However, few studies have addressed the relationship between MGD and hormone replacement therapy (HRT) in perimenopausal women, and not have examined the prevalence of MGD in perimenopausal women. The purpose of this study was to address and evaluate the possible relationship between MGD and HRT in perimenopausal women.The results suggest that perimenopausal women have a high prevalence of DE related to perimenopausal symptoms. The study also shows that perimenopausal women who use HRT can gain benefits for DE as well as for perimenopausal symptoms. Physicians caring for women who are experiencing DE related to perimenopausal symptoms should consider HRT.

Differential effects of low- and high-dose glucocorticoids on the innate immunity of corneal epithelium in vitro.

PURPOSE: To evaluate the effect of glucocorticoids on the innate immunity of corneal epithelial cells. METHODS: Human corneal epithelial cell lines (HCEC) were co-cultured with LPS and hydrocortisone to determine whether hydrocortisone modulates the expression and function of TLR2, 4. The release of IL-6, 8 from cultured HCEC was measured in the presence and absence of specific blocking antibodies to TLR2, 4. The proteins of TLR2, 4 were also compared by Western blot. RESULTS: Incubation of HCEC with LPS upregulated the expression of TLR2, 4 and increased the release of IL-6, 8. This upregulation was enhanced by low-concentration hydrocortisone, but inhibited by high-concentration hydrocortisone. The concentration of IL-6, 8 was also enhanced by low-concentration hydrocortisone and inhibited by high-concentration hydrocortisone. CONCLUSIONS: Low-concentration hydrocortisone enhances the expression and function of TLRs in HCEC and provides evidence for a novel function of glucocorticoids in innate immunity.

High glucose may decrease the innate immune through TLRs in cornea epithelium.

PURPOSE: The purpose of this study was to investigate the high potential of glucose in inhibiting the innate immune in cultured human cornea epithelial cells (HCEC) and try to determine whether the role of high glucose on the HCEC relate to toll-like receptor (TLR)2 and TLR4. METHODS: Cells were cultured for 3 days in 5 mmol/l (normal glucose). Then high glucose (25 mmol/l) was added along with normal glucose with daily changes in media for 24 h. The cells were also treated with mannitol as an osmotic control. The cellular abundance of the mRNAs for TLR2 and TLR4 was determined by real-time polymerase chain reaction analysis. The proteins of TLR2 and TLR4 were also compared by immunofluorescent staining and western blot. The release of interleukin 6 (IL-6) and IL-8 from cultured HCEC was measured using enzyme-linked immunosorbent assays (ELISA) in the presence and absence of specific blocking antibodies to TLR2 and TLR4. RESULTS: Incubation of HCEC with high glucose showed that the mRNA expression of TLR2 and TLR4 was markedly inhibited. Immunofluorescent staining and western blot analysis confirmed that the protein expression of TLR2 and TLR4 was downregulated in response to high glucose. The result of ELISA also showed that the release of IL-6 and IL-8 can be inhibited by high glucose, but these inhibitions were partly counteracted after pretreatment with anti-TLR2 and/or anti-TLR4 monoclonal antibody. The results also showed that the osmotic control did not affect the expression of TLR2, TLR4, and IL-6, 8. CONCLUSIONS: High glucose may decrease the innate immune through TLRs in cornea epithelium.

Hydrocortisone suppression of the expression of VEGF may relate to toll-like receptor (TLR) 2 and 4.

PURPOSE: The purpose of this study was to investigate hydrocortisone's potential in inhibiting the expression of VEGF in cultured human cornea fibroblasts (HCF) and to determine whether the role of hydrocortisone in the expression of VEGF may relate to toll-like receptors 2 and 4 (TLR2 and TLR4). METHODS: Three different concentrations of hydrocortisone were used to stimulate the cultured HCF. The cellular abundance of the mRNAs for VEGF and TLR2, 4 were determined by real-time polymerase chain reaction analysis. The release of IL-6, IL-8, and VEGF from cultured HCF was measured using enzyme-linked immunosorbent assays (ELISA) in the presence and absence of specific blocking antibodies to TLR2, 4. The proteins of TLR2, 4 were also compared by Western blot. RESULTS: Following incubation of HCF with hydrocortisone, we found the mRNA expression of TLR2, 4 and VEGF were markedly inhibited. ELISA and Western blot analysis confirmed that protein expression of TLR2, 4 and VEGF was down-regulated in response to hydrocortisone. The result of ELISA also showed the release of IL-6 and IL-8 can be inhibited by hydrocortisone. But all these inhibitions were partly counteracted after pretreatment with anti-TLR2 and/or anti-TLR4 monoclonal antibody. CONCLUSIONS: Hydrocortisone may decrease the expression of VEGF through inhibiting TLR2, 4 activity in cultured human corneal fibroblasts.

Modulation of expression of Toll-like receptors in the human endometrium.

PROBLEM: The purpose of the study was to investigate the different expressions of toll-like receptors (TLRs) in the proliferative and the secretory phase of endometrial tissue. METHOD OF STUDY: Eight infertile women were included in this study. The endometrial tissues of proliferative and secretory phase were obtained from each woman. The tissues were evaluated for the expression of mRNA for TLR1-10 by reverse transcribed polymerase chain reaction (RT-PCR). The proteins of TLR2, 3, 4, and 9 were evaluated by Western blot. The mRNA and protein level of proliferative and secretory endometrial tissue from the same woman was compared. The data were analysed with SPSS15.0. RESULTS: TLR1-7, 9, and 10 mRNA were expressed throughout the menstrual cycle, but in the same woman, the expression of TLR2-6, 9, and 10 mRNA was higher during the secretory phase than that in the proliferative phase. The Western blot also showed that the protein expression of TLR2, 3, 4, and 9 was stronger in the secretory phase than that in the proliferative phase in the same woman. CONCLUSION: The expression of TLRs is cycle dependent in human endometrial tissue. The expressions of TLRs were higher in the secretory phase than that in the proliferative phase: this indicated that TLRs may be regulated by sex hormones throughout the menstrual cycle.

Effects of a cyclooxygenase-1-selective inhibitor in a mouse model of ovarian cancer, administered alone or in combination with ibuprofen, a nonselective cyclooxygenase inhibitor.

Nonsteroidal anti-inflammatory drugs (NSAIDs) are known to be potent inhibitors of the cyclooxygenases. The present study was designed to investigate the effects of a cyclooxygenase (COX)-1 inhibitor, SC-560, administered alone or in combination with ibuprofen on the growth inhibition of s.c. human ovarian SKOV-3 carcinoma and on angiogenesis. The effects of SC-560 and ibuprofen on tumor growth inhibition have been examined in mouse ovarian cancer models. Angiogenesis of both COX inhibitors was measured by reverse-transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. Prostaglandin E(2) (PGE(2)) levels in tumor tissues of mice were also determined by ELISA. The inhibitory rates in SC-560 group alone and in combination with ibuprofen group were 21.21% and 41.55%, respectively. In combination therapy with SC-560 and ibuprofen, tumor volumes were significantly reduced compared with that of control group (P < 0.05). In treatment groups, both COX inhibitors significantly reduced intratumor PGE(2) levels (all P < 0.01). Microvessel density (MVD) in tumor tissues were significantly decreased from 80.90 +/- 5.14 in vehicle-treated to 40.70 +/- 10.45 and 38.90 +/- 8.41 in SC-560 group alone and combination ibuprofen therapy (all P < 0.01). Ibuprofen was similar to the cyclooxygenase-1-selective inhibitor SC-560 in its ability to suppress the values of MVD of tumor tissues. SC-560 administered alone or in combination with ibuprofen inhibited the COX-associated up-regulation of VEGF. These studies demonstrate synergism between two COX inhibitors and that antiangiogenic therapy can be used to inhibit ovarian cancer growth.

Expression of toll-like receptors in the Fusarium solani infected cornea.

PURPOSE: To investigate the expression of toll-like receptors (TLRs) in the human cornea with Fusarium solani (F. solani) keratitis. METHODS: Five human corneas with F. solani keratitis and 5 healthy human corneas were evaluated for TLR1-10 mRNA by reverse transcribed polymerase chain reaction (RT-PCR) and relative real-time PCR. The proteins of TLR2, 4, and 9 were also compared by Western blot. The mean times between these corneas were collected, and the onset of treatment to surgery was 34.4 +/- 12.4 days, ranging from 15 to 47 days. The data were analyzed with SPSS15.0. RESULTS: TLR1-10 mRNA was expressed in both healthy and F. solani infected human corneas. The expression of TLR2, 4, 6, and 9 mRNA in the F. solani infected human corneas were upregulated. The Western blot showed the protein expression of TLR2, 4, and 9 was also upregulated in the corneas with F. solani keratitis than that of the healthy corneas. CONCLUSIONS: TLRs are expressed diversely in the F. solani infected human cornea, and TLR2, 4, and 9 may be implicated in the pathogenesis of F. solani infection in the cornea.

[Improvement of cervical microenvironment after treatment of chronic cervicitis with focused ultrasound in infertility women].

OBJECTIVE: To investigate the effects of focused ultrasound (FUS) treatment on cervical microenvironment in infertility women with chronic cervicitis. METHODS: A total of 10 women treated in Infertility Clinic of Obstetrics and Gynecology Hospital of College of Medicine of Zhejiang University were assessed. The women aged from 25 to 34 with normal menstrual cycles had moderate or severe cervical erosion. Ultrasound showed they had normal ovulation. The sperms of their husbands had normal motility and number. The FUS they received had a focal depth range of 4 - 6 mm, a resonant frequency of 9 - 11 MHz, and an output power of 3.5 W. The follow-up time was three months. The changes of cervical mucus and the results of postcoital test were evaluated. The clinical effects on cervical erosion and complications were also evaluated. RESULT: In 6 cases of severe cervical erosion, 2 were cured, 2 were improved significantly (more than 50 % reduction of erosion area) and 2 were improved (less than 50 % reduction of erosion area). In 4 cases of moderate cervical erosion, 2 were cured, 2 were improved significantly. After the treatment, the median of cervical mucus score during LH peak were increased from 11 to 13 (Wilcoxon test, P=0.014). Postcoital test showed that the median of highly motile sperm (grade III and grade II) of 5 high-power field were increased from 22 to 52 (Wilcoxon test, P=0.015). Mild side effects without medical intervention included vaginal fluid in all 10 women, vaginal spotting in 2 women. No serious side effects were observed. CONCLUSION: The focused ultrasound therapy can help to improve the cervical microenvironment for infertility women with moderate or severe cervical erosion without serious side effects.