[Role of the liver-enriched transcription factor binding site mutation in the C promoter region of hepatitis B virus genome for HBx-enhanced hepatitis B virus replication].

Objective: To construct a recombinant HBV replication-type plasmid with liver-enriched transcription factor binding site mutation at proximal of HBV C promoter in order to elucidate the role of HBx-enhanced HBV replication. Methods: Site-directed mutagenesis technology was used to construct a recombinant plasmid with liver-enriched transcription factor binding site mutation at proximal of HBV C promoter on the basis of wild-type HBV replicating plasmid and HBV replicating plasmid lacking HBx expression. Subsequently, plasmid transfection was carried out in HBV liver cancer cell replication model and mouse replication model, and HBV replication intermediates of cells and mouse liver tissue were extracted for detection. Results: Based on the HBV replicating plasmid, the HBV replicating plasmid with liver-enriched transcription factor binding site mutation at proximal of HBV C promoter was successfully constructed. HBx-enhanced HBV replication were detected in both the HBV liver cancer replication model and the mouse replication model. After mutating liver-enriched transcription factor binding site mutation at proximal of HBV C promoter, the effect of HBx on the enhancement of HBV replication was not significantly affected. Conclusion: HBx may not enhance HBV replication through liver-enriched transcription factor binding site mutation at proximal of HBV C promoter. The role of other liver-enriched transcription factor binding sites in HBx-enhanced HBV replication needs further study.

[Clinical approach and prognosis of continuous neurological care after intensive care for the patients with severe and refractory anti-N-methyl-D-aspartate receptor encephalitis].

Objective: To study the clinical features, continuous care and prognosis of the patients with severe and refractory anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis after intensive care unit (ICU). Methods: Clinical data of patients with severe and refractory anti-NMDAR encephalitis, who were transferred from ICU to general ward of neurology between December 2015 and October 2019, were retrospectively reviewed and analyzed in the study. Results: Twenty patients (11 females and 9 males) were enrolled in the study. The median course of disease when patients were transferred to general ward was 4.4 (2.0, 6.0) months. Six cases were alert, 6 cases were in a coma, 5 were in the early recovery phase and 3 were in the late recovery phase. Severe malnutrition, pneumonia, urinary tract infections, bedsores and leukocytopenia were common complications. Seven out of 18 patients were tested positive for cerebrospinal fluid anti-NMDAR antibodies with high titers (≥1∶100). During this continuous therapy stage,10 patients were treated with intravenous immunoglobulin (IVIg), 1 with methylprednisolone, 2 with rituximab, 1 with intrathecal methotrexate and 1 received intravenous cyclophosphamide. All Patients were prescribed a long-term immunotherapy (mycophenolate mofetil 1.5-3.0 g/d). Sixteen patients (80%) had good prognosis (modified Rankin Scale (mRS)≤2), and the mortality was 10%, with follow-up time of 17.0 (8.0, 27.0) months. Conclusions: Patients with anti-NMDAR encephalitis, who are transferred from ICU, have severely impaired neurologic function. These patients need long-term individualized immunotherapy and continuous neurological care. Good outcomes can be achieved in most patients.

Circular RNA hsa_circ_0008039 promotes proliferation, migration and invasion of breast cancer cells through upregulating CBX4 via sponging miR-515-5p.

OBJECTIVE: Breast cancer (BC) is the second most frequent malignancy worldwide. Hsa_circ_0008039 exerts the carcinogenic factors in BC. However, the pathogenesis of hsa_circ_0008039 involved in BC is still unclear. PATIENTS AND METHODS: The expression levels of hsa_circ_0008039, microRNA-515-5p (miR-515-5p) and chromobox homolog 4 (CBX4) in BC tissues and cells were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Cell proliferation, migration and invasion were assessed by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) and transwell assays, severally. The binding relationship among hsa_circ_0008039, miR-515-5p and CBX4 was predicted by starBase, then verified by the dual-luciferase reporter assay and immunoprecipitation (RIP) assay. The interaction between hsa_circ_0008039 and miR-515-5p was confirmed by RNA pull-down assay. The protein level of CBX4 was detected by Western blot assay. The biological role of hsa_circ_0008039 was detected by xenograft tumor model in vivo. RESULTS: Hsa_circ_0008039 was upregulated in BC tissues and cells, and expedited proliferation, migration and invasion of BC cells. MiR-515-5p was downregulated in BC tissues and cells and worked as a target of hsa_circ_0008039. CBX4 was highly expressed in BC tissues and cells, and contributed to proliferation, migration and invasion of BC cells. Hsa_circ_0008039 enhanced CBX4 expression by competitively binding to miR-515-5p, thereby promoting BC development. Hsa_circ_0008039 knockdown repressed BC tumor growth in vivo. CONCLUSIONS: These findings implicated that hsa_circ_0008039 contributed to proliferation, migration and invasion in vitro and promoted tumor growth in vivo by miR-515-5p/CBX4 axis in BC, suggesting a potential therapeutic strategy for BC treatment.

LncRNA ROR1-AS1 promotes colon cancer cell proliferation by suppressing the expression of DUSP5/CDKN1A.

OBJECTIVE: The purpose of this study was to explore the possible role of ROR1-AS1 in the pathogenesis of colon cancer and the underlying mechanism. PATIENTS AND METHODS: The expression levels of ROR1-AS1 in 75 colon cancer tissue samples and adjacent ones, as well as in cell lines were examined by quantitative Polymerase Chain Reaction (qPCR). Then, ROR1-AS1 overexpression plasmid and siRNA were transfected into colon cancer cells using liposome method. After that, Cell Counting Kit-8 (CCK-8) and plate colony formation assays were conducted to analyze cell proliferation, while flow cytometry was applied for the analysis of cell cycle and apoptosis. At last, the mechanism of action of ROR1-AS1 was further explored by nuclear separation, RNA binding protein immunoprecipitation (RIP) and chromatin immunoprecipitation (CHIP) assays. RESULTS: ROR1-AS1 level in colon cancer tissues was remarkably higher than that in normal tissues, and the expression in tumors of stage III and IV was remarkably higher than those of stage I and II. Meanwhile, tumors with diameters more than 5 cm had a higher ROR1-AS1 expression than those less than 5 cm. After transfection with ROR1-AS1 overexpression plasmid, the cell proliferation ability was enhanced, the G0/G1 phase time of cell cycle was shortened, and the apoptosis was suppressed. However, the opposite result was observed after ROR1-AS1 was downregulated. Furthermore, RIP showed that ROR1-AS1 can bind to enhancer of zeste homolog 2 (EZH2) and inhibit the expression of DUSP5, and thus be engaged in the proliferation and apoptosis of colon cancer cells. CONCLUSIONS: ROR1-AS1 is highly expressed either in colon cancer tissues or in cell lines, which is able to enhance cell proliferation, accelerate cell cycle, and inhibit cell apoptosis. The mechanism of ROR1-AS1 to participate in the development of colon cancer may be the downregulation of DUSP5 via combination with EZH2.

LINC01605 promotes the proliferation of laryngeal squamous cell carcinoma through targeting miR-493-3p.

OBJECTIVE: The purpose of this study was to elucidate the potential influence of LINC01605 on the progression of laryngeal squamous cell carcinoma (LSCC) and the underlying mechanism. PATIENTS AND METHODS: LINC01605 and microRNA-493-3p (miR-493-3p) levels in normal laryngeal tissues, LSCC tissues, and paired paracancerous tissues were detected. Regulatory effects of LINC01605 on proliferative ability and apoptosis in HEp-2 and AMC-HN-8 cells were assessed. Besides, the interaction between LINC01605 and miR-493-3p was evaluated by Dual-Luciferase reporter gene assay and Spearman's rank correlation analysis. Finally, rescue experiments were conducted to clarify the role of LINC01605/miR-493-3p axis in the progression of LSCC. RESULTS: LINC01605 was upregulated and miR-493-3p was downregulated in LSCC tissues. Knockdown of LINC01605 inhibited proliferative ability, and stimulated apoptosis in HEp-2 and AMC-HN-8 cells. Moreover, LINC01605 directly bound to miR-493-3p, and the former negatively regulated the level of the latter. In addition, miR-493-3p was able to reverse the regulatory effect of LINC01605 on proliferative ability in LSCC. CONCLUSIONS: LINC01605 is upregulated in LSCC tissues, and it promotes the malignant progression of LSCC via targeting miR-493-3p.

[Analysis of the way of thinking on pharmacology in Lu Zhiyi's Bencao Chengya Banji].

More than 410 kinds of prescriptions of medicines were presented, some of which were collected in Shennong Bencao Jing(, Shennong's Classic of Materia Medica) and other herbal works, are included in Lu Zhiyi's Bencao Chengya Banji(). He introduced the indications of these medicines by focusing on the name of the herbs, or the species of them, and the features and growth morphology of the herbs. He abstracted the efficacy of drugs on the human body. Therefore, he noted the indications of all the medicines list in the book.

BRAF mutation correlates with recurrent papillary thyroid carcinoma in Chinese patients.

PURPOSE: We investigated correlations of somatic BRAF V600E mutation and RET/PTC1 rearrangement with recurrent disease in Chinese patients with papillary thyroid carcinoma (ptc). METHODS: This prospective study included 214 patients with ptc histologically confirmed between November 2009 and May 2011 at a single institute. RESULTS: We found somatic BRAF V600E mutation in 68.7% and RET/PTC1 rearrangement in 25.7% of the patients. Although BRAF mutation was not significantly associated with clinicopathologic features such as patient sex or age, multicentric disease, thyroid capsule invasion, tumour stage, or nodal metastasis, it was significantly associated with recurrent disease. Multivariate analysis revealed that BRAF mutation and tumour size were independent risk factors associated with recurrent disease, with odds ratios of 9.072 and 2.387 respectively. The area under the receiver operating characteristic curve increased 8.3% when BRAF mutation was added to the traditional prognostic factors, but that effect was statistically nonsignificant (0.663 vs. 0.746, p = 0.124). RET/PTC1 rearrangement and nodal metastasis were significantly associated in all patients (p = 0.042), marginally associated in ptc patients (p = 0.051), but not associated in microptc patients (p = 0.700). RET/PTC1 rearrangement was not significantly associated with recurrent disease. CONCLUSIONS: BRAF positivity is an independent predictor of recurrent disease in ptc.

Androgen receptor inclusions acquire GRP78/BiP to ameliorate androgen-induced protein misfolding stress in embryonic stem cells.

Commitment of differentiating embryonic stem cells (ESCs) toward the various lineages is influenced by many factors, including androgens. However, the mechanisms underlying proteotoxic stress conferred by androgen receptor (AR) actions on embryonic cell fate remains unclear. Here we show that mouse ESCs display stress-related cellular phenotypes in response to androgens during early phase of differentiation. Androgen induced a significant increase in the percentage of ESCs and embryoid bodies with the intranuclear and juxtanuclear AR inclusions, which were colocalized with the E3 ubiquitin ligase, C terminus of Hsc70-interacting protein. Caspase-3 activity corresponded with AR expression, was enhanced in cells engaged more differentiation phenotypes. Androgen-mediated accumulation of AR aggregates exacerbated endoplasmic reticulum (ER) stress and rendered ESCs susceptible to apoptosis. Increasing expression levels of the ER chaperones, GRP78/BiP and GRP94, as well as ER stress markers, such as ATF6, phosphorylated PERK, GADD153/CHOP and spliced XBP-1 mRNA, were dramatically elevated in ESCs overexpressing AR. We found that androgen induced GRP78/BiP to dissociate from ATF6, and act as an AR-interacting protein, which was recruited into AR inclusions in ESCs. GRP78/BiP was also colocalized with AR inclusions in the cells of spinal bulbar muscular atrophy transgenic mouse model. Overexpression of GRP78/BiP suppressed ubiquitination of AR aggregates and ameliorated the misfolded AR-mediated cytopathology in ESCs, whereas knockdown of GRP78/BiP increased the accumulation of AR aggregates and significantly higher levels of caspase-3 activity and cell apoptosis. These results generate novel insight into how ESCs respond to stress induced by misfolded AR proteins and identify GRP78/BiP as a novel regulator of the AR protein quality control.

Clinical feature analysis of fatal pulmonary thromboembolism: experiences from 41 autopsy-confirmed cases.

AIM: Due to non-specific symptoms and imaging features, a timely and accurate diagnosis of pulmonary thromboembolism (PTE) is often difficult. This study aims to evaluate the frequency of, and risk factors for, autopsy-confirmed cases with fatal pulmonary thromboembolism (FPE) that were missed or misdiagnosed before death. MATERIAL AND METHODS: Forensic autopsies that were performed at the Center of Forensic Medicine in West China were retrospectively reviewed, and demographic and clinical data of autopsy-confirmed cases with FPE were collected. RESULTS: There were 41 cases with pathologically confirmed FPE, which represents 7.3% (41/558) of autopsy cases that documented sudden death in hospital. Of those 41 cases, only 14.6% (6/41) were correctly diagnosed before death, and 85.4% (35/41) were missed or misdiagnosed. According to medical records, bowel movements and out-of-bed activity were the major triggers of FPE death, and 90% of cases had at least two of the known risk factors for PTE. Increasing age, orthopedic surgery, and multiple traumas were the most common risk factors. Additionally, of the 41 cases with FPE, 51.2% (21/41) died in the Orthopedic Department. CONCLUSIONS: FPE was common in older patients who had a recent history of surgery and multiple traumas. Increasing the early diagnosis of PTE in high-risk patients may be useful for reducing the incidence of FPE.

Effect of retinoic acid on implantation and post-implantation development of mouse embryos in vitro.

BACKGROUND: This study was designed to examine the embryotoxic potential of retinoic acid (RA) at the blastocyst stage and during early post-implantation development of mouse embryos in vitro. METHODS AND RESULTS: All-trans retinoic acid (t-RA) was administered to ICR mice embryos at a dose level of 0, 0.001 micromol/l, 0.1 micromol/l and 10 micromol/l throughout in-vitro culture. A total of 404 embryos was randomly assigned to all different dose groups. The percentage of embryos in later stages of development changed depending upon the dose of RA used. Exposure to 10 micromol/l of t-RA at the blastocyst stage, implanted blastocyst stage or early oocyte stage was also found to cause different degrees of retardation of embryo development and embryo death. These findings demonstrate, for the first time, that RA exerts an adverse effect on embryo growth during the early post-implantation stages of development, in comparison with day 3 to day 8 of gestation in vivo. CONCLUSIONS: Although isotretinoin (13-cis-retinoic acid) is effective for the therapy of cystic acne and other dermatological disorders, retinoid treatment should be avoided at the early post-implantation stage of gestation.

Determination of the efficiency of controlled ovarian hyperstimulation in the gonadotropin-releasing hormone agonist-suppression cycle using the initial follicle count during gonadotropin stimulation.

PURPOSE: Our purpose was to evaluate the relationship between the initial follicle count during gonadotropin stimulation after gonadotropin-releasing hormone (GnRH) agonist suppression and the efficiency of controlled ovarian hyperstimulation (COH) in patients receiving treatment with assisted reproductive technologies (ARTs). METHODS: A total of 338 COH procedures in 291 couples was performed with cycles that reached the stage of oocyte retrieval. The ovarian antral follicle number was measured using transvaginal ultrasonography at the folliculometry during gonadotropin stimulation by GnRH agonist suppression in patients undergoing ARTs. Controlled ovarian hyperstimulation was accomplished using GnRH agonist down-regulation combined with FSH and menotropin stimulation. The characteristics of oocytes after retrieval and embryos after in vitro culture and the pregnancy rates were assessed. RESULTS: The procedures performed included 195 ET cycles, 129 TET cycles, and 14 incomplete cycles. The treatment cycles were divided into four categories according to the antral follicle number (i.e., < or = 5, 6-10, 11-15, and > or = 16) at the first folliculometry to evaluate the influence of various factors. The antral follicle count correlated significantly with the patient age, dosage of gonadotropins, serum estradiol concentration, number of antral follicles (> or = 13 mm) while receiving hCG injections, number of oocytes retrieved, and, later, number of embryos transferred. There was a trend toward an increasing number of pregnancies per cycle as the number of antral follicles increased (14.7, 26.5, 44, and 45%, respectively). CONCLUSIONS: We were able to predict the efficiency of COH and outcome of ARTs based on the follicle count during the first folliculometry during gonadotropin stimulation after GnRH agonist suppression. The results of the folliculometry significantly predicted the ovarian response to COH and the outcome of ARTs in the current treatment cycle.

Effects of retinoic acid on pre-implantation embryo development in mice.

BACKGROUND: In a previous study we had investigated the effect of in utero retinoic acid (RA) exposure on early post-implantation development at the blastocyst stage before implantation and immediately after implantation to understand the possible roles of RA in embryogenesis. The results showed that excess RA affected early post-implantation embryogenesis adversely. We designed the present study to investigate the effect of in utero RA exposure on pre-implantation embryos. METHODS: In the prospective animal study, pregnant female mice received early pre-implantation peanut oil with 50 mg/kg t-RA or 100 mg/kg t-RA by oral gavage on the morning of day 1 and 2 or late pre-implantation exposure on the night of day 2 and morning of day 3 of gestation. Mice were sacrificed late in day 3. The number and morphology embryos were recorded. RESULTS: All mice given oral RA were sacrificed on the same day. The mean number of embryos per mouse and the percentage of different embryo stages in the t-RA treated mice administered at early pre-implantation embryo or late pre-implantation embryo stage were not significantly different from the controls. The mean number of embryos per 50 mg/kg mouse and the percentage of expanded blastocysts or early blastocysts/morulas did not differ from controls. In addition, the percentage of expanded blastocysts or early blastocysts/morulas was also not significantly different from the control group in 100 mg/kg late pre-implantation mice. CONCLUSION: Mouse embryo development may not experience dose related adverse effects from non-physiological RA exposure during pre-implantation.

Effects of induction anesthetic agents on outcome of assisted reproductive technology: a comparison of propofol and thiopental sodium.

BACKGROUND: The use of propofol, as compared to barbiturates (e.g. thiopental), for short surgical procedures has been associated with more rapid recovery from the procedure. Propofol, an intravenous anesthetic drug, is frequently used as an adjunct to transvaginal oocyte retrieval but little is known about its effects upon fertilization, embryo development, and pregnancy rate when used drug the for induction of general anesthesia. This study was conducted to compare the outcome of assisted reproductive technology (ART) with the use of propofol versus thiopental sodium for the induction of general anesthesia during oocyte retrieval. METHODS: In this retrospective study, 92 cases of infertile patients who underwent oocyte retrieval under the induction of general anesthesia with, respectively, either propofol (Group I, 72 cases) or thiopental sodium (Group II, 20 cases) were compared for fertilization rate, cleavage rate, and pregnancy rate. RESULTS: The fertilization rate was 68.9% for Group I and 66.7% for group II (p = 0.614). The cleavage rate was 96.5% for Group I and 94.8% for Group II (p = 0.294). The rate of good embryo grading and poor embryo grading was, respectively, 85.1% and 14.9% for Group I, versus 85.7% and 14.3% for Group II (p = 0.887). The pregnancy rate was 30.5% for Group I and 20.0% for Group II (p = 0.354). The implantation rate and abortion rate was, respectively, 9.1% and 18.2% for Group I versus 7.2% and 25.0% for Group II (implantation rate, p = 0.590; abortion rate, p = 0.600). CONCLUSION: There were no significant differences between these two groups for fertilization rate, cleavage rate, pregnancy rate, implantation rate and abortion rate. We suggest cautious use of propofol for the procedure of oocyte retrieval despite its associated more rapid post-operative recovery including less nausea/vomiting.

Two different timings of intrauterine insemination for non-male infertility.

PURPOSE: Our purpose was to assess the simplicity and convenience of treatment scheduled not on weekends, by comparing two different timings of intrauterine insemination (IUI) protocol. METHODS: A prospective observational study of two different protocols of intrauterine insemination was designed. Two hundred and ten infertile couples with normal spermiograms were included in this study. Fifty-eight couples were treated with IUI 26 to 28 h after human chorionic gonadotropin (hCG) injection plus timed intercourse within a 12- to 18-hr period and 147 couples had IUI 36 to 38 hr after hCG injection and timed intercourse within a 12- to 18-hour period. Pregnancy rates were compared with two different protocols of IUI. RESULTS: The mean age, duration, and causes of infertility and the cycle characteristics following follicular stimulation were similar between the two groups. The cycle characteristics of follicular stimulation in the two treatment groups were not different. There also were no significant differences between the groups in the type of sperm concentration, sperm motility, and the percentage of sperm with normal morphology per insemination. The number of follicles greater than 17 mm per patient was not significantly different between the two groups. The pregnancy rate per cycle also was similar between the two groups in men with lower motile sperm numbers (< 40 x 10(6)) (23.6% vs. 23.4%) and in men with higher sperm numbers (> or = 40 x 10(6)) (25% vs. 24.4%). CONCLUSIONS: The different timing but similar efficacy of these two IUI protocols provides a practical choice to clinicians. The availability of both protocols may avoid unnecessary scheduling of clinical and laboratory work on weekends and holidays in women participating in controlled ovarian hyperstimulation and IUI programs for treatment of non-male infertility.

Seminal plasma zinc levels and sperm motion characteristics in infertile samples.

BACKGROUND: Zinc (Zn) in seminal plasma stabilizes the cell membrane and nuclear chromatin of spermatozoa. It may also have an antibacterial function. However, extremely high concentrations of Zn (10 to 100 x the normal range) may inhibit sperm motility and the function of the mannose receptor on the sperm head. In this study, we analyzed the correlation between Zn levels in seminal plasma and the characteristics of semen as measured by conventional and computer aided sperm analysis (CASA). METHODS: One hundred fifteen infertile couples were recruited for conventional semen analysis and CASA from December 1995 through January 1996, and Zn levels in semen samples were determined by flame atomic absorption spectroscopy (AAS). RESULTS: A good correlation in a positive direction (r = 0.73, p = 0.0001) was noted between the total amount of Zn per ejaculate and the Zn concentration. The Zn concentration in seminal plasma was negatively correlated with the seminal pH (r = -0.35, p = 0.0081). There was no significant correlation between the total amount of Zn per ejaculate and sperm characteristics, including sperm count, motility (% sperm count), progressive motility (% motility), rapid motility (% motility), average path velocity (VAP, microns/s), straight-line velocity (VSL, microns/s), curvilinear velocity (VCL, microns/s), amplitude of lateral head displacement (ALH, microns), beat/cross frequency (BCF, beats/s), straightness (STR), and linearity (LIN). There was also no significant correlation between the Zn concentration in seminal plasma and the above sperm characteristics. CONCLUSION: The characteristics of semen as determined by conventional semen analysis or CASA bore no correlation with total seminal Zn amount or Zn concentrations in the ejaculates. Routine determination of the Zn concentration in seminal plasma offers no advantages in infertility work-ups.

Comparable clinical outcomes of tubal embryo transfer for oligoastheno-teratozoospermia treated with intracytoplasmic sperm injection and for female infertility treated with in vitro fertilization.

BACKGROUND: Female and male indications may each have their negative impacts on the success of assisted reproductive technologies. Reports regarding the outcomes of in vitro fertilization (IVF) vs. intracytoplasmic sperm injection (ICSI) are controversial. This study was conducted to investigate whether the clinical outcome of tubal embryo transfer (TET) for oligoastheno teratozoospermia treated with ICSI is different from that of tubal embryo transfer for female infertility treated with IVF. METHODS: From January 1997 to December 1998, results of tubal embryo transfers of 54 IVF (IVF-TET) cycles for female infertility were retrospectively compared with those of 49 ICSI (ICSI-TET) cycles for oligoastheno-teratozoospermia. RESULTS: The fertilization rates were 78.1% +/- 21.8%, and 78.0% +/- 21.9% for treatments with IVF-TET and ICSI-TET, respectively. The implantation rates were 13.8% and 21.2%, respectively. The pregnancy rates per transfer were 46.2% and 48.9%, and the abortion rates were 14.8% and 12.2% for treatments with IVF-TET and ICSI-TET, respectively. The above rates for the IVF-TET and ICSI-TET groups were comparable. CONCLUSION: Male infertility due to oligoastheno teratozoospermia treated with ICSI-TET appears to have a comparable outcome to female infertility treated with IVF-TET.

Clinical implications of intracytoplasmic sperm injection using cryopreserved testicular spermatozoa from men with azoospermia.

OBJECTIVE: To investigate whether sperm obtained by testicular sperm extraction (TESE) and cryopreserved well before intracytoplasmic sperm injection (ICSI) can serve as an effective sperm source. STUDY DESIGN: The role of cryopreserved testicular spermatozoa was evaluated in a retrospective analysis of consecutive ICSI cycles using fresh or cryopreserved sperm; they were followed by prospective, planned treatment using cryopreserved sperm with a modified ICSI procedure. Sixteen men (22 cycles) with obstructive or nonobstructive azoospermia were included in the retrospective analysis. Another 25 men (29 cycles) were in the planned treatment group. Following these series, the pregnancy outcomes were compared between ICSI cycles with fresh or cryopreserved testicular sperm. RESULTS: In the retrospective phase, 14 ICSI cycles were performed using fresh sperm, with 8 using cryopreserved sperm. There were no statistically significant differences between the two groups in any outcome measure. Planned treatment with cryopreserved sperm resulted in a fertilization rate of 84% and an embryo transfer rate of 89%. Thirteen couples (44%) achieved pregnancy (five ongoing, six delivered). These rates were similar to those in the retrospective phase of the study. All couples in the planned cryopreservation group had multiple aliquots (6.5 +/- 2.1) of sperm remaining after the first cycle. CONCLUSION: Cryopreserved sperm obtained by TESE can be used as an effective sperm source in ICSI cycles. Planned cryopreservation allows multiple aliquots to be stored for use in subsequent cycles and thus avoids the need for repeat biopsies.

Influence of polyvinylpyrrolidone on the outcome of intracytoplasmic sperm injection.

OBJECTIVE: To assess the effectiveness of a procedure for intracytoplasmic sperm injection (ICSI) modified so as not to use polyvinylpyrrolidone (PVP) and to examine clinical outcome. STUDY DESIGN: Seventy-seven cycles of ICSI were performed over a one-year period. PVP was used for sperm immobilization in 39 of these cycles and was eliminated from the other 38 cycles. Difference in fertilization rate, cleavage rate, parthenogenetic activity, clinical pregnancy rate, ongoing pregnancy rate and grading of preembryos between the two groups was compared. RESULTS: The non-PVP group had a higher fertilization rate (57.63% vs. 84.43%, P < .001) and better preembryo quality (chi 2 = 6.80, P = .009) than the PVP group. There was no significant difference in cleavage rate, parthenogenetic activity, clinical pregnancy rate and ongoing pregnancy rate between the two groups. CONCLUSION: Performing ICSI without PVP may improve the fertilization rate and preembryo grading. However, further study with a larger cohort is necessary to determine whether the modified procedure can increase the pregnancy rate.

The expression of DAZL1 in the ovary of the human female fetus.

OBJECTIVE: To determine whether DAZL1 is expressed in human fetal ovarian tissue. DESIGN: The presence of DAZL1 expression was determined by reverse transcriptase polymerase chain reaction (RT-PCR). SETTING: Academic tertiary care medical center and research unit of university. PATIENT(S): Five female abortuses between the 19th and 22nd week of gestational age. INTERVENTION(S): Fetal ovarian tissues were collected immediately after the cessation of the heart beat. MAIN OUTCOME MEASURE(S): The product of RT-PCR. RESULT(S): DAZL1 expression was detected in all five samples. CONCLUSION(S): DAZL1 is not only expressed in human testes but also in ovaries. It may play a role in germ cell survival and gonad development in both sexes.

Multiple pregnancy with adnexal torsion after in vitro fertilization: case report.

Assisted reproductive techniques (ART) are widely accepted procedures for infertile couples. Rare complications, like heterotopic pregnancy, bilateral tubal pregnancy, and adnexal torsion during pregnancy, have been diagnosed with increasing frequency after ART. We present a case of an early triplet pregnancy complicated with adnexal torsion. The patient was pregnant through in vitro fertilization. Early ultrasound examination revealed a triplet pregnancy within the uterine cavity. At 7 weeks' gestational age, an acute onset of lower abdominal pain, progressive abdominal distension, and massive internal bleeding prompted emergency laparotomy. The right ovary was enlarged, twisted, necrotic and hemorrhagic. Attempts to preserve the ovary failed because of the friable nature of the affected ovary, and an oophorectomy had to be performed. Although the removed ovary contained a corpus luteum, the pregnancy continued smoothly after only short luteal support. A precise pre-surgery diagnosis in our case was difficult based on the patient's initial clinical presentation. However, with high clinical suspicion in addition to color Doppler ultrasound, the physician should be able to make an early decision for an exploratory laparotomy or laparoscopy, gaining the benefit of more conservative treatment.