Identification and characterization of camptothecin tailoring enzymes in Nothapodytes tomentosa.

Camptothecin is a complex monoterpenoid indole alkaloid with remarkable antitumor activity. Given that two C-10 modified camptothecin derivatives, topotecan and irinotecan, have been approved as potent anticancer agents, there is a critical need for methods to access other aromatic ring-functionalized congeners (e.g., C-9, C-10, etc.). However, contemporary methods for chemical oxidation are generally harsh and low-yielding when applied to the camptothecin scaffold, thereby limiting the development of modified derivatives. Reported herein, we have identified four tailoring enzymes responsible for C-9 modifications of camptothecin from Nothapodytes tomentosa, via metabolomic and transcriptomic analysis. These consist of a cytochrome P450 (NtCPT9H) which catalyzes the regioselective oxidation of camptothecin to 9-hydroxycamptothecin, as well as two methyltransferases (NtOMT1/2, converting 9-hydroxycamptothecin to 9-methoxycamptothecin), and a uridine diphosphate-glycosyltransferase (NtUGT5, decorating 9-hydroxycamptothecin to 9-ß-D-glucosyloxycamptothecin). Importantly, the critical residues that contribute to the specific catalytic activity of NtCPT9H have been elucidated through molecular docking and mutagenesis experiments. This work provides a genetic basis for producing camptothecin derivatives through metabolic engineering. This will hasten the discovery of novel C-9 modified camptothecin derivatives, with profound implications for pharmaceutical manufacture.

An E3 ubiquitin ligase CSIT2 controls critical sterility-inducing temperature of thermo-sensitive genic male sterile rice.

Thermo-sensitive genic male sterile (TGMS) lines are the core of two-line hybrid rice (Oryza sativa). However, elevated or unstable critical sterility-inducing temperatures (CSITs) of TGMS lines are bottlenecks that restrict the development of two-line hybrid rice. However, the genes and molecular mechanisms controlling CSIT remain unknown. Here, we report the CRITICAL STERILITY-INDUCING TEMPERATURE 2 (CSIT2) that encodes a really interesting new gene (RING) type E3 ligase, controlling the CSIT of thermo-sensitive male sterility 5 (tms5)-based TGMS lines through ribosome-associated protein quality control (RQC). CSIT2 binds to the large and small ribosomal subunits and ubiquitinates 80S ribosomes for dissociation, and may also ubiquitinate misfolded proteins for degradation. Mutation of CSIT2 inhibits the possible damage to ubiquitin system and protein translation, which allows more proteins such as catalases to accumulate for anther development and inhibits abnormal accumulation of reactive oxygen species (ROS) and premature programmed cell death (PCD) in anthers, partly rescuing male sterility and raised the CSIT of tms5-based TGMS lines. These findings reveal a mechanism controlling CSIT and provide a strategy for solving the elevated or unstable CSITs of tms5-based TGMS lines in two-line hybrid rice.

Perceptions of transitional care services among patients with percutaneous transhepatic biliary drainage and multicentre health professionals: A qualitative study.

BACKGROUND: Patients with percutaneous transhepatic biliary drainage (PTBD) need regular drainage tube care after discharge, and transitional care can help solve this problem. However, few studies have focused on the quality of transitional care, the perceptions of patients with drainage tubes after discharge and those of healthcare professionals. AIM: This study is aimed at exploring the real experience and perceptions of transitional care services among healthcare professionals and PTBD patients who have been discharged with tubes and at providing references for future transitional care service development. DESIGN: The study uses a qualitative descriptive design. The reporting method followed Consolidated Criteria for Reporting Qualitative Research guidelines. METHODS: Semistructured interviews were conducted with PTBD patients who had been discharged with tubes and multicentre healthcare professionals using the purpose sampling method. The thematic analysis method was used for analysis. RESULTS: Thirteen PTBD patients from one hospital and 12 healthcare professionals from three hospitals were interviewed. The analysis of the patient interview data revealed three themes, namely, recognition of the value of transitional care services, patients have some unmet needs and perception of transitional care service pathways. Six subthemes were also identified. The analysis of the interview data of healthcare professionals revealed two themes, namely, harvest and challenges in transitional care services work and expectations for future development of transitional care services. Four subthemes were also identified. CONCLUSIONS: The transitional care of discharged patients with PTBD tubes deserves the attention of clinical workers, and a series of measures should be taken to improve transitional care services. PATIENT/PUBLIC CONTRIBUTION: Patients were involved in the formulation of interview questions for this study, and during the interviews, patients presented their suggestions for transitional care services. Healthcare professionals participated in this study as interviewees, and no members of the public were involved in this study.

The E3 ubiquitin ligase CSIT1 regulates critical sterility-inducing temperature by ribosome-associated quality control to safeguard two-line hybrid breeding in rice.

Two-line hybrid breeding can fully utilize heterosis in crops. In thermo-sensitive genic male sterile (TGMS) lines, low critical sterility-inducing temperature (CSIT) is vital to safeguard the production of two-line hybrid seeds in rice (Oryza sativa), but the molecular mechanism determining CSIT is unclear. Here, we report the cloning of CSIT1, which encodes an E3 ubiquitin ligase, and show that CSIT1 modulates the CSIT of thermo-sensitive genic male sterility 5 (tms5)-based TGMS lines through ribosome-associated quality control (RQC). Biochemical assays demonstrated that CSIT1 binds to the 80S ribosomes and ubiquitinates abnormal nascent polypeptides for degradation in the RQC process. Loss of CSIT1 function inhibits the possible damage of tms5 to the ubiquitination system and protein translation, resulting in enhanced accumulation of anther-related proteins such as catalase to suppress abnormal accumulation of reactive oxygen species and premature programmed cell death in the tapetum, thereby leading to a much higher CSIT in the tms5-based TGMS lines. Taken together, our findings reveal a regulatory mechanism of CSIT, providing new insights into RQC and potential targets for future two-line hybrid breeding.

BOTRYOID POLLEN 1 regulates ROS-triggered PCD and pollen wall development by controlling UDP-sugar homeostasis in rice.

Uridine diphosphate (UDP)-sugars are important metabolites involved in the biosynthesis of polysaccharides and may be important signaling molecules. UDP-glucose 4-epimerase (UGE) catalyzes the interconversion between UDP-Glc and UDP-Gal, whose biological function in rice (Oryza sativa) fertility is poorly understood. Here, we identify and characterize the botryoid pollen 1 (bp1) mutant and show that BP1 encodes a UGE that regulates UDP-sugar homeostasis, thereby controlling the development of rice anthers. The loss of BP1 function led to massive accumulation of UDP-Glc and imbalance of other UDP-sugars. We determined that the higher levels of UDP-Glc and its derivatives in bp1 may induce the expression of NADPH oxidase genes, resulting in a premature accumulation of reactive oxygen species (ROS), thereby advancing programmed cell death (PCD) of anther walls but delaying the end of tapetal degradation. The accumulation of UDP-Glc as metabolites resulted in an abnormal degradation of callose, producing an adhesive microspore. Furthermore, the UDP-sugar metabolism pathway is not only involved in the formation of intine but also in the formation of the initial framework for extine. Our results reveal how UDP-sugars regulate anther development and provide new clues for cellular ROS accumulation and PCD triggered by UDP-Glc as a signaling molecule.

Characterization of two putative norlaudanosoline methyltransferases from Aristolochia debilis.

In view of the nephrotoxicity, hepatotoxicity, and carcinogenicity of aristolochic acids (AAs), the removal of AAs from plants becomes an urgent priority for ensuring the safety of Aristolochia herbal materials. In this study, based on the root-predominant distribution of aristolochic acid I (AAI) in Aristolochia debilis, transcriptome sequencing, in combination with phylogenetic analyses, and gene expression pattern analysis together provided five candidate genes for investigating AAI biosynthesis. Comprehensive in vitro and in vivo enzymatic assays revealed that Ab6OMT1 (6-O-methyltransferase) and AbNMT1 (N-methyltransferase) exhibit promiscuity in substrate recognition, and they could act in a cooperative fashion to achieve conversion of norlaudanosoline, a predicted intermediate in AAI biosynthetic route, into 3'-hydroxy-N-methylcoclaurine through two different methylation reaction sequences. These results shed light on the molecular basis for AAI biosynthesis in Aristolochia herbs. More importantly, Ab6OMT1 and AbNMT1 may be employed as targets for the metabolic engineering of AAI biosynthesis to produce AAs-free Aristolochia herbal materials.

O-Alkylazoxymycins A-F, Naturally Occurring Azoxy-Aromatic Compounds from Streptomyces sp. Py50.

Six new azoxy-aromatic compounds (o-alkylazoxymycins A-F, 1-6) and two new nitrogen-bearing phenylvaleric/phenylheptanoic acid derivatives (o-alkylphemycins A and B, 7 and 8) were isolated from Streptomyces sp. Py50. Their structures were elucidated based on HRESIMS, NMR, UV spectroscopic analyses, and X-ray crystallographic data. O-Alkylazoxymycins A-F (1-6) are the first natural examples of azoxy compounds with the azoxy bond attached to the ortho-position of the phenylheptanoic acid or phenylvaleric acid moiety. Compounds 1, 5, and 6 were active against Epidermophyton floccosum with MIC50 values ranging from 10.1 to 51.2 µM. A plausible biosynthetic pathway of 2 and 3 was proposed.

Environment-sensitive genic male sterility in rice and other plants.

Environment-sensitive genic male sterility is a type of male sterility that is affected by both genetic and environmental factors. Environment-sensitive genic male sterile lines are not only used in two-line hybrid breeding but are also good materials for studying plant-environment interactions. In this study we review the research progress on environment-sensitive genic male sterility in rice from the perspectives of epigenetic, transcriptional, posttranscriptional, posttranslational and metabolic mechanisms as well as signal transduction processes. While significant progress has been made in the genetics, gene cloning and understanding of the molecular mechanisms of environment-sensitive genic male sterility in recent years, the relevant regulatory network is still poorly understood in rice. We therefore also review studies of environment-sensitive genic male sterility in Arabidopsis and other crops, hoping to promote research in this field in rice. Finally, we analyse the challenges posed by environment-sensitive genic male sterility and provide corresponding suggestions. This review will contribute towards an understanding the molecular genetics of environment-sensitive genic male sterility and its application in two-line hybrid breeding in rice and other species.

Application of a new body surface-assisting puncture device in percutaneous transforaminal endoscopic lumbar discectomy.

BACKGROUND: Accurate puncture and localization are critical for percutaneous transforaminal endoscopic lumbar discectomy surgery. However, several punctures are often required, followed by X-ray fluoroscopy, which can increase surgical risk and complications. The aim of this study was to demonstrate a new body surface-assisting puncture device that can be used in percutaneous transforaminal endoscopic lumbar discectomy and to assess its clinical effectiveness. METHODS: Three hundred and forty-four patients were treated with percutaneous transforaminal endoscopic lumbar discectomy surgery in the Spinal Surgery Department of Taian City Central Hospital, China, between January 2020 and February 2022. Of these, 162 patients (the locator group) were punctured using a body surface-assisting puncture device while and 182 patients (the control group) were punctured using the traditional blind puncture method. The number of punctures, radiation dose during X-ray fluoroscopy, operation time, and surgical complications were compared between the two groups. RESULTS: The average number of punctures was 2.15 ± 1.10 in the locator group which was significantly lower than that in the control group (5.30 ± 1.74; P < 0.001). The average X-ray fluoroscopy radiation dose in the locator group was significantly lower at 2.34 ± 0.99 mGy, compared with 5.13 ± 1.29 mGy in the control group (P < 0.001). The mean operation time was also significantly less in locator group (47.06 ± 5.12 vs. 62.47 ± 5.44 min; P = 0.008). No significant differences in surgical complications were found between the two groups (P > 0.05). CONCLUSION: The use of a new body surface-assisting puncture device in percutaneous transforaminal endoscopic lumbar discectomy surgery can significantly reduce the number of punctures and X-ray fluoroscopy radiation dose, as well as shortening the operation time, without increasing surgical complications. This device is cheap, easy to operate, and suitable for all hospitals and spine surgeons, especially for small hospitals, with also no extra costs for patients.

Comparison of the safety and efficacy of hepatic resection and radiofrequency ablation in the treatment of single small hepatocellular carcinoma: systematic review and meta-analysis.

Background: The present study aimed to compare the safety and efficacy of minimally invasive hepatic resection (HR) and radiofrequency ablation (RFA) in the treatment of single small hepatocellular carcinoma via systematic review and meta-analysis. Methods: We conducted electronic literature searches of PubMed, Embase, and The Cochrane Library databases, A case-controlled trial comparing HR and RFA in the treatment of single small hepatocellular carcinoma published between 2010 and January 15, 2022 was searched and reported outcomes were overall survival, postoperative complications, intraoperative blood loss, duration of surgery, and recurrence. Literature met inclusion criteria were screened out, and the quality of the methodology used in the included literature was evaluated using the Newcastle-Ottawa Scale (NOS). Meta-analysis was performed using RevMan 5.4 software. Results: Eleven articles were included, including 2,001 patients, including 1,071 in the RFA group and 930 in the HR group. Offset risk assessment results are published offset. Meta-analysis showed that the overall survival rate at 1, 3, and 5 years in RFA group was higher than that in HR group, the difference between the two groups was statistically significant at 1 year [odds ratio (OR) =1.57, 95% confidence interval (CI): 1.14, 2.17, P=0.006], 3 years (OR =1.81, 95% CI: 1.42, 2.30, P<0.00001], and 5 years (OR =1.87, 95% CI: 1.47, 2.37, P<0.00001). The incidence of postoperative complications was significantly lower in the RFA group than in the human resources group [risk ratio (RR) =1.75, 95% CI: 1.02, 3.00, P=0.04], and length of hospital stay [standard mean difference (SMD) =2.92, 95% CI: 0.54, 5.30, P=0.02], operation time (SMD =2.87, 95% CI: 2.57, 3.16, P<0.00001) were shorter than those in HR group. However, the recurrence rate of RFA group was higher than that of HR group (OR =0.49, 95% CI: 0.36, 0.66, P<0.00001). Conclusions: RFA has the advantage of having more advantages and fewer complications in the treatment of small hepatocellular carcinoma. Minimal complications can occur for patients when achieving satisfactory treatment results. A new treatment option is available for clinicians in the treatment of small liver cancer.

Methylesterification of cell-wall pectin controls the diurnal flower-opening times in rice.

Flowers are the core reproductive organ of plants, and flowering is essential for cross-pollination. Diurnal flower-opening time is thus a key trait influencing reproductive isolation, hybrid breeding, and thermostability in plants. However, the molecular mechanisms controlling this trait remain unknown. Here, we report that rice Diurnal Flower Opening Time 1 (DFOT1) modulates pectin methylesterase (PME) activity to regulate pectin methylesterification levels of the lodicule cell walls, which affect lodicule swelling to control diurnal flower-opening time. DFOT1 is specifically expressed in the lodicules, and its expression gradually increases with the approach to flowering but decreases with flowering. Importantly, a knockout of DFOT1 showed earlier diurnal flower opening. We demonstrate that DFOT1 interacts directly with multiple PMEs to promote their activity. Knockout of PME40 also resulted in early diurnal flower opening, whereas overexpression of PME42 delayed diurnal flower opening. Lower PME activity was observed to be associated with higher levels of pectin methylesterification and the softening of cell walls in lodicules, which contribute to the absorption of water by lodicules and cause them to swell, thus promoting early diurnal flower opening. Higher PME activity had the opposite effect. Collectively, our work uncovers a molecular mechanism underlying the regulation of diurnal flower-opening time in rice, which would help reduce the costs of hybrid breeding and improve the heat tolerance of flowering plants by avoiding higher temperatures at anthesis.

The structural characteristics and the substrate recognition properties of RNase ZS1.

TMS5 encodes an RNase ZS1 protein that can process ubiquitin-60S ribosomal protein L40 family (UbL40) mRNAs to regulate thermo-sensitive genic male sterility in rice. Despite the importance of this protein, the structural characteristics and substrate recognition properties of RNase ZS1 remain unclear. Here, we found that the variations in several conservative amino acids alter the activation of RNase ZS1, and its recognition of RNA substrates depends on the structure of RNA. RNase ZS1 acts as a homodimer. The conserved amino acids in or adjacent to enzyme center play a critical role in the enzyme activity of RNase ZS1 and the conserved amino acids that far from active center have little impact on its enzyme activity. The cleavage efficiency of RNase ZS1 for pre-tRNA-MetCAU35 and UbL401 mRNA with cloverleaf-like structure was higher than that of pre-tRNA-AspAUC9 and UbL404 mRNA with imperfect cloverleaf-like structure. This difference implies that the enzyme activity of RNase ZS1 depends on the cloverleaf-like structure of the RNA. Furthermore, the RNase ZS1 activity was not inhibited by the 5' leader sequence and 3' CCA motif of pre-tRNA. These findings provide new insights for studying the cleavage characteristics and substrate recognition properties of RNase ZS.

Ubiquitinome Profiling Reveals the Landscape of Ubiquitination Regulation in Rice Young Panicles.

Ubiquitination, an essential post-transcriptional modification (PTM), plays a vital role in nearly every biological process, including development and growth. Despite its functions in plant reproductive development, its targets in rice panicles remain unclear. In this study, we used proteome-wide profiling of lysine ubiquitination in rice (O. sativa ssp. indica) young panicles. We created the largest ubiquitinome dataset in rice to date, identifying 1638 lysine ubiquitination sites on 916 unique proteins. We detected three conserved ubiquitination motifs, noting that acidic glutamic acid (E) and aspartic acid (D) were most frequently present around ubiquitinated lysine. Enrichment analysis of Gene Ontology (GO) annotations and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of these ubiquitinated proteins revealed that ubiquitination plays an important role in fundamental cellular processes in rice young panicles. Interestingly, enrichment analysis of protein domains indicated that ubiquitination was enriched on a variety of receptor-like kinases and cytoplasmic tyrosine and serine-threonine kinases. Furthermore, we analyzed the crosstalk between ubiquitination, acetylation, and succinylation, and constructed a potential protein interaction network within our rice ubiquitinome. Moreover, we identified ubiquitinated proteins related to pollen and grain development, indicating that ubiquitination may play a critical role in the physiological functions in young panicles. Taken together, we reported the most comprehensive lysine ubiquitinome in rice so far, and used it to reveal the functional role of lysine ubiquitination in rice young panicles.

Molecular Control and Application of Male Fertility for Two-Line Hybrid Rice Breeding.

The significance of the climate change may involve enhancement of plant growth as well as utilization of the environmental alterations in male fertility (MF) regulation via male sterility (MS) systems. We described that MS systems provide a fundamental platform for improvement in agriculture production and have been explicated for creating bulk germplasm of the two-line hybrids (EGMS) in rice as compared to the three-line, to gain production sustainability and exploit its immense potential. Environmental alterations such as photoperiod and/or temperature and humidity regulate MS in EGMS lines via genetic and epigenetic changes, regulation of the noncoding RNAs, and RNA-metabolism including the transcriptional factors (TFs) implication. Herein, this article enlightens a deep understanding of the molecular control of MF in EGMS lines and exploring the regulatory driving forces that function efficiently during plant adaption under a changing environment. We highlighted a possible solution in obtaining more stable hybrids through apomixis (single-line system) for seed production.

High-Expression of Neuropilin 1 Correlates to Estrogen-Induced Epithelial-Mesenchymal Transition of Endometrial Cells in Adenomyosis.

Epithelial-mesenchymal transition (EMT) induced by estrogen contributes to the development of adenomyosis. However, the exact underlying mechanism remains mostly obscure. We hypothesized that a transmembrane glycoprotein neuropilin 1 (NRP1) was critical in the EMT induced by estrogen, accelerating the development of adenomyosis. We firstly investigated the expression pattern of NRP1 in endometrium samples from women with adenomyosis. We found that NRP1 expression was significantly increased in the endometrium of uterine adenomyosis, especially in the ectopic endometrium. To determine the role of NRP1 in the EMT in endometrial cells, we used an NRP1 overexpression retrovirus to up-regulate the NPR1 expression in human endometrial cells (HEC-1-A). Endometrial cells infected with NRP1 retroviruses showed a high expression of NRP1 and exerted a mesenchymal phenotype, characterized by down-regulation of E-cadherin and Occludin, up-regulation of α-SMA and N-cadherin, and enhanced migration. Then, we found that 17ß-estradiol (E2) up-regulated the expression of NRP1 in endometrial cells in a dose-dependent manner, which was eliminated by raloxifene, a selective estrogen receptor inhibitor. Importantly, NRP1 shRNA significantly suppressed the EMT induced by E2 in endometrial cells. And NRP1 shRNA significantly inhibited the phosphorylation of Smad3 and restored the expressions of Slug and Snail1 mRNA. Collectively, these data highlight the possible role of NRP1 in the EMT in the development of adenomyosis and provide a potential therapeutic target for adenomyosis patients.

HMS1 interacts with HMS1I to regulate very-long-chain fatty acid biosynthesis and the humidity-sensitive genic male sterility in rice (Oryza sativa).

Environment-sensitive genic male sterility (EGMS) lines are used widely in two-line hybrid breeding in rice (Oryza sativa). At present, photoperiod-sensitive genic male sterility (PGMS) lines and thermo-sensitive genic male sterility (TGMS) lines are predominantly used in two-line hybrid rice, with humidity-sensitive genic male sterility (HGMS) lines rarely being reported. Here, it is shown that HUMIDITY-SENSITIVE GENIC MALE STERILITY 1 (HMS1), encoding a ß-ketoacyl-CoA synthase, plays key roles in the biosynthesis of very-long-chain fatty acids (VLCFAs) and HGMS in rice. The hms1 mutant displayed decreased seed setting under low humidity, but normal seed setting under high humidity. HMS1 catalyzed the biosynthesis of the C26 and C28 VLCFAs, contributing to the formation of bacula and tryphine in the pollen wall, which protect the pollen from dehydration. Under low-humidity conditions, hms1 pollen showed poor adhesion and reduced germination on the stigmas, which could be rescued by increasing humidity. HMS1-INTERACTING PROTEIN (HMS1I) interacted with HMS1 to coregulate HGMS. Furthermore, both japonica and indica rice varieties with defective HMS1 exhibited HGMS, suggesting that hms1 potentially could be used in hybrid breeding. The results herein reveal the novel mechanism of VLCFA-mediated pollen wall formation, which protects pollen from low-humidity stress in rice, and has a potential use in hybrid crop breeding.

Purification and characterization of a bioactive alpha-fetoprotein produced by HEK-293 cells.

Alpha-fetoprotein (AFP) is a biomarker that is used to diagnose hepatocellular carcinoma (HCC) and can promote malignancy in HCC. AFP is an important target in the treatment of liver cancer. To obtain enough AFP to screen for AFP inhibitors, we expressed and purified AFP in HEK-293 cells. In the present study, we produced AFP in the cells and harvested highly pure rAFP (or recombinant expression AFP in HEK-293 cells). We also analysed the bioactivity of rAFP and found that rAFP promoted growth of the human HCC cells, antagonize paclitaxel inhibition of HCC cell proliferation, suppress expression of active caspase-3, and promote expression of Ras and survivin. This study provides a method to produce significant amounts of AFP for use in biochemical assays and functional studies and to screen AFP inhibitors for use in HCC therapy.

[EZH2 gene silenced by siRNA suppresses the growth and invasion of endometrial carcinoma cells].

OBJECTIVE: To explore the effects on cell proliferation and invasion as well as molecular basis after suppressing EZH2 expression in endometrial carcinoma cells by using siRNAs. METHODS: RT-PCR was used to examine the expression of EZH2 in endometrial carcinoma and their paracancerous tissues. SiRNAs targeting to EZH2 were transfected to endometrial carcinoma cells, and MTT, FACS, and boyden assays were utilized to examine cell proliferation, cell cycle change, and cell invasion. Finally, the molecular mechanisms of EZH2 on cell function alteration were investigated. RESULTS: Compared with paracancerous tissues, increased expression trend of EZH2 mRNA was showed in endometrial carcinoma tissues. Further, knocking down EZH2 expression inhibited cell growth, cell cycle transition from G1 to S phase, and cell invasion ability. Molecular basis indicated that suppression of EZH2 downregulated the expression of E2F1 and MMP9 and upregulated tumor suppressor p21 expression. CONCLUSION: EZH2 expression is increased in endometrial carcinoma tissues. Knocking down EZH2 expression suppresses the cell growth, cell cycle transition and cell invasion by downregulated E2F1 and MMP9, and upregulated tumor suppressor p21 expression.

[Effect of formaldehyde on expressions of Fas apoptosis pathway-related genes of ovary tissues in female rats].

OBJECTIVE: To explore the mechanism of formaldehyde inducing ovarian toxicity in female rats by observing the effect of formaldehyde on the expression of Fas and caspase-8 mRNA, and the activity of caspase-3 and caspase-8 of ovary tissues in female rats. METHODS: Forty female Sprague-Dawley(SD) rats were randomly divided into a control group and 3 formaldehyde groups at different concentrations. The rats in the formaldehyde groups were intraperitoneally injected different doses of formaldehyde (20.0,2.0 and 0.2 mg/kg) continuously for 14 days.After 14 days, all rats were sacrificed and their ovaries were collected for detecting the expression of Fas and caspase-8 mRNA with RT-PCR, the protein expression of Fas with Western blot, and the activities of caspase-8 and caspase-3 with spectrophotometric method. RESULTS: Compared with the control group, the expression of Fas mRNA and its protein and caspase-8 mRNA and the activity of caspase-8 and caspase-3 of ovary tissues in the rats treated with formaldehyde significantly increased with dose (P<0.05). CONCLUSION: The increase of Fas gene expression and the activity of caspase-8 and caspase-3 may be the important mechanism of ovarian toxicity induced by formaldehyde in female rats.

[Clinical observation of young, middle-aged and elderly women with endometrial carcinoma].

OBJECTIVE: To explore the clinical features, diagnosis, treatment and prognosis of endometrial carcinoma in young, middle-aged and elderly women. METHODS: We retrospectively analyzed the clinical data of 82 cases of endometrial carcinoma in young, middle-aged women and 33 cases of endometrial cacinoma in elderly women. RESULTS: The rates of adenocarcinoma in young, middle-aged and elderly groups were 74.4% and 75.5%, respectively. The young,middle-aged and elderly patients with Stage I endometrial cancer were 64.6% and 69.7%, and those with Stage III and IV were 15.9% and 15.2%, respectively. The histological Grade 1 carcinoma of endometrium in young,middle-aged and elderly women were 70.7% and 60.6%, respectively. The young, middle-aged women without myometrial invasion were more than the elderly women (42.8% vs 15.6%, P < 0.01). The young, middle-aged women with myometrial invasion more than half of myometrial wall were less than the elderly women (10.4% vs 40.6%, P < 0.01). The rate of chemical treatment after the surgery in the elderly women was more than that of the young, middle-aged women (P < 0.05). The 5-year survival rate of the young, middle-aged women was obviously higher than that of the elderly women (92.79% vs 72.21%, P < 0.05). CONCLUSION: Adenocarcinoma and well-differentiated cells are the main pathological characteristics of endometrial carcinoma both in the young, middle-aged and the elderly women. Most young, middle-aged and el-derly patients can be diagnosed and treated in the early stage. Early diagnosis and reasonable treatment can improve the prognosis. The prognosis of the young, middle-aged patients is obviously better than that of the elderly patients, and the myometrial invasion depth may be the main difference.