Alocasia macrorrhiza Represents a New Host of 'Candidatus Phytoplasma asteris'-Related Strains Associated with Yellows Symptoms in China.

Alocasia macrorrhiza, which belongs to the Araceae family, is an important landscape plant in China, and has of significant medicinal uses. In 2022, A. macrorrhiza displaying abnormal symptoms were found in Qionghai, Hainan Island of China (110°23'3.06″，19°7'56.29″). The incidence of symptomatic plants was about 40% in the sampled areas. The abnormal symptoms included that the ovoid leaves color turned yellow from green gradually, with ovoid leaves chlorosis, mesophyll tissue yellowing, miniature leaves and systemic wilting. The diseased symptoms suspected to be associated with phytoplasma according to the protocols of phytoplasma identification. In order to identify the pathogen, eleven diseased samples and three asymptomatic samples were collected from an area of about 40 hectares. Total DNAs were extracted from 0.10 g fresh plant leaf tissues using a CTAB DNA extraction method. PCR amplifications were performed using primers R16mF2/R16mR1 and fTuf1/rTuf1 specific for the phytoplasma 16S rRNA and tuf genes. Target PCR amplicons were obtained from the DNA of 11 diseased samples, whereas not from the DNA of the asymptomatic samples. The PCR products were cloned and sequenced by Biotechnology (Shanghai) Co., Ltd. (Guangzhou, China), and the obtained sequences were assembled, edited and analyzed using the EditSeq program and DNAMAN version 6.0. The phytoplasma 16S rRNA and tuf gene amplicons were 1336 and 930 bp in length, respectively. The sequences of all 16S rRNA and tuf amplicons in this study were identical. The sequencing data were deposited in GenBank with accession numbers OR466206 (16S rDNA) and OR513090 (tuf). According to the methods and protocols of phytoplasma identified and classification, the phytoplasma strain was described as Alocasia macrorrhiza yellows (AmY) phytoplasma, AmY-hn strain. BLAST search were conducted based on 16Sr RNA and tuf genes. The results showed that the AmY-hn had 100 % 16Sr RNA sequence identity (1336 bp out of 1336 bp) with that of 16SrI-B subgroup phytoplasmas like onion yellows phytoplasma (OY-M, AP006628). The AmY-hn had 100 % tuf sequence identity (930 bp out of 930 bp) with that of 16SrI-B subgroup phytoplasmas like OY-M. RFLP profiles obtained with iPhyClassifier demonstrated that AmY-hn strain was a member of the 16SrI-B subgroup with a similarity coefficient 1.00 to the reference phytoplasma strain (AP006628). Separated phylogenetic analysis based on 16S rRNA and tuf genes obtained with MEGA 7.0 using the neighbor-joining (NJ) method with 1000 bootstrap value indicated that AmY-hn clustered into one clade with phytoplasma strains of OY-M and chinaberry witches'-broom (KP662119) with 100 % and 87 % bootstrap value respectively. To our knowledge, this is the first report that a 'Candidatus Phytoplasma asteris'-related strain belonging to 16SrI-B subgroup infects A. macrorrhiza in China. The 16SrI-B subgroup 'Candidatus Phytoplasma asteris'-related strains can spread outwards through the plant A. macrorrhiza. Thus, the findings in the study will be beneficial to the detection of phytoplasmas which parasitic in this plant and the epidemic monitoring of the related diseases.

Molecular Identification of 'Candidatus Phytoplasma malaysianum'-Related Strains Associated with Areca catechu Palm Yellow Leaf Disease and Phylogenetic Diversity of the Phytoplasmas within 16SrXXXII Group.

Areca catechu palm is an important cash plant in Hainan Island of China and even tropical regions worldwide. Areca catechu palm yellow leaf (AcYL) disease caused by the phytoplasmas is a devastating disease for the plant production. In the study, the phytoplasmas associated with the AcYL diseases were identified and characterized based on the conserved genes of the phytoplasmas, and genetic variation and phylogenetic relationship of the phytoplasma strains in the 16SrXXXII group was demonstrated. The results indicated that Areca catechu palm showing yellow leaf symptoms were single infected by 'Candidatus Phytoplasma malaysianum'-related strains belonging to 16SrXXXII-D subgroup. BLAST and multiple sequence alignment analysis based on 16S rRNA and secA genes showed that the AcYL phytoplasmas shared 100% sequence identity and 100% homology with the 'Ca. Phytoplasma malaysianum'-related strains. Phylogenetic analysis indicated that the AcYL phytoplasmas and 'Ca. Phytoplasma malaysianum'-related strains belonging to 16SrXXXII group were clustered into one clade with a 100% bootstrap value. Based on computer-simulated digestions, 6 kinds of RFLP patterns within 16SrXXXII group were obtained and a novel subgroup in the 16Sr group was recommended to propose to describe the relevant strains in this 16Sr subgroup. To our knowledge, this is the first report that Areca catechu palm showing yellow leaf symptoms infected by 'Ca. Phytoplasma malaysianum'-related strains belonging to 16SrXXXII group. And a novel 16Sr subgroup 16SrXXXII-F was proposed based on the systematical analysis of genetic variation of all the phytoplasmas within 16SrXXXII group. The findings of this study would support references for monitoring the epidemiology and developing effective prevention strategies of the AcYL diseases.

Molecular Identification and Characterization of Two Groups of Phytoplasma and Candidatus Liberibacter Asiaticus in Single or Mixed Infection of Citrus maxima on Hainan Island of China.

The pathogens associated with citrus Huanglongbing symptoms, including yellowing and mottled leaves in Citrus maxima, an important economic crop on Hainan Island of China, were identified and characterized. In the study, detection, genetic variation and phylogenetic relationship analysis of the pathogens were performed based on 16S rRNA and ß-operon gene fragments specific to phytoplasma and Candidatus Liberibacter asiaticus. The results indicated that the pathogens-such as phytoplasma strains of CmPII-hn belonging to the 16SrII-V subgroup and CmPXXXII-hn belonging to the 16SrXXXII-D subgroup, as well as Candidatus Liberibacter asiaticus strains CmLas-hn-were identified in the diseased plant samples, with numbers of 12, 2 and 6 out of 54, respectively. Among them, mixed infection with the 16SrII-V subgroup phytoplasma and Candidatus Liberibacter asiaticus was found in the study, accounting for 7.4% (four samples). The phytoplasma strains of CmPII-hn-Tephrosia purpurea witches' broom, Melochia corchorifolia witches' broom and Emilia sonchifolia witches' broom-were clustered into one clade belonging to the 16SrII-V subgroup, with a 99% bootstrap value. The phytoplasma strains of CmPXXXII-hn and Trema tomentosa witches' broom belonging to 16SrXXXII-D, and the other 16SrXXXII subgroup strains were clustered into one clade belonging to the 16SrXXXII group with a 99% bootstrap value. There were 16 variable loci in the 16S rRNA gene sequences of the tested 16SrXXXII group phytoplasma strains, of which two bases had an insertion/deletion. The strains of Candidatus Liberibacter asiaticus, identified in the study and the strains that had been deposited in GenBank, were in one independent cluster with a 99% bootstrap value. To our knowledge, this is the first report showing that Citrus maxima can be infected by 16SrII-V and16SrXXXII-D subgroup phytoplasmas in China. Moreover, this is also the first report in which the plants are co-infected by 16SrII-V subgroup phytoplasmas and Candidatus Liberibacter asiaticus. More comprehensive and detailed identification and characterization of the pathogens associated with the diseased symptoms in Citrus maxima on the island in China would be beneficial for epidemic monitoring and for the effective prevention and control of related plant diseases.

Carica papaya Represents a New Host of 16SrI-B Subgroup Phytoplasma Associated with Yellow Symptoms in China.

Carica papaya Linn, belonging to the Caricaceae family, is an economic and medicinal plant, which is widely cultivated in tropical and subtropical countries (Soib et al., 2020). Beginning in 2021, abnormal symptoms of Carica papaya exhibiting leaf yellow, crinkle and leaflet were found in Wanning city of Hainan Province, China. The diseased symptoms of the plant, with about 20 % incidence in the sampling regions, were suspected to be induced by phytoplasma, a phloem-limited and could not be cultured in vitro prokaryotic pathogen. Total DNAs were extracted from 0.10 g fresh leaves of symptomatic or asymptomatic Carica papaya using CTAB DNA extraction method (Doyle and Doyle, 1990). PCR reactions were performed using primers R16mF2/R16mR1 (Gundersen and Lee, 1996), secAfor1/secArev3 (Hodgetts et al., 2008) and AYgroelF/AYgroelR (Mitrovic et al., 2011) specific for phytoplasma 16S rRNA, secA and groEL gene fragments. PCR products of the 16S rRNA, secA and groEL gene target fragments of phytoplasma were obtained from the DNA of eight diseased Carica papaya samples whereas not from the DNA of the asymptomatic plant samples. The PCR amplicons of the three genes were cloned and sequenced by Biotechnology (Shanghai) Co., Ltd. (Shanghai, China) and the sequences data were deposited in GenBank. The 16S rRNA, secAgroEL gene of phytoplasma was in length of 1326 (GenBank accession: OL625608), 716 (OL630087) and 1300 (OL630088) bp separately, putatively encoding 238 (secA) and 432 (groEL) amino acids sequence. The phytoplasma strain was named as Carica papaya yellow phytoplasma (CpY), CpY-hnwn strain. A blast search based on 16Sr RNA gene of CpY-hnwn showed 100 % sequence identity with that of 16SrI aster yellows group members (16SrI-B subgroup), such as Onion yellows phytoplasma strain OY-M (AP006628), Chinaberry witches'-broom phytoplasma (CWB) strain CWB-hnsy1 (KP662119) and CWB-hnsy2 (KP662120), Rapeseed phyllody phytoplasma isolate RP166 (CP055264). RFLP analysis based on the 16S rRNA gene fragment of CpY-hnwn was performed by the interactive online phytoplasma classification tool iPhyClassifier (Zhao et al., 2009) indicated that the phytoplasma strain is a member of 16SrI-B subgroup. Blast search based on secA gene of CpY-hnwn showed 100 % sequence identity with that of CWB strains CWB-gdgz (KP662182), CWB-jxnc (KP662180) and CWB-fjya (KP662178) belonging to 16SrI-B subgroup. Blast search based on groEL gene of CpY-hnwn showed 99.77 % sequence identity with that of mulberry dwarf phytoplasma (AB124809) and 99.69 % sequence identity with that of Onion yellows phytoplasma strain OY-M (AP006628) and Rapeseed phyllody phytoplasma isolate RP166 (CP055264). Phylogenetic analysis based on the 16S rRNA gene fragments performed by MEGA 7.0 employing neighbor-joining (NJ) method with 1000 bootstrap value (Kumar et al., 2016; Felsenstein, 1985) indicated that the CpY-hnwn phytoplasma strain was clustered into one clade with the phytoplasma strains of OY-M (AP006628), RP166 (CP055264), CWB-hnsy1 (KP662119), CWB-hnsy2 (KP662120) and areca palm yellow leaf (KF728948), with 100 % bootstrap value. To our knowledge, this is the first report that a 16SrI-B subgroup phytoplasma infects Carica papaya in Hainan Province, a tropical island of China. Carica papaya was previously reported to be infected by 16SrXII-O subgroup phytoplasmas in Nigeria (Kazeem et al., 2021), 16SrII-U subgroup in Hainan Province of China (Yang et al., 2016). The findings in this study indicated that one plant couldthe phytoplasmas belonging to different 16Sr groups, which would be beneficial to the specific detection of the pathogens and the epidemic monitoring of the related diseases. References: Doyle, J.J. and Doyle, J.L. 1990. Focus 12: 13-15. Felsenstein, J. 1985. Evolution 39: 783-791. Gundersen, D.E. and Lee, I.M. 1996. Phytopath. Medit. 35: 144-151. Hodgetts, J., et al. 2008. Int. J. Syst. Evol. Microbiol. 58: 1826-1837. Kazeem, S.A., et al. 2021. Crop Prot. 148: 105731. Kumar, S., et al. 2016. Mol. Biol. Evol. 33: 1870-1874. Mitrovic, J., et al. 2011. Ann. Appl. Biol. 159: 41-48. Soib, H.H., et al. 2020. Molecules, 25: 517. Yang, Y., et al. 2016. Int. J. Syst. Evol. Microbiol. 66: 3485-3491. Zhao, Y., et al. 2009. Int. J. Syst. Evol. Microbiol. 59: 2582-2593.

Utilization and expenses of outpatient services among tuberculosis patients in three Chinese counties: an observational comparison study.

BACKGROUND: The China-Gates TB project Phase II implemented case-based payment reform in three Chinese counties in 2014, designed specifically for patients diagnosed with Tuberculosis (TB). This study aimed to examine the changes in utilization and expenses of outpatient services before and after the reform implementation, among TB patients in the three counties in China. METHODS: We collected quantitative data using surveys in 2013 (baseline year) and 2015 (final year). We used outpatient hospital records to measure service utilization and medical expenses of TB patients. We conducted qualitative interviews with local health authorities, officers of health insurance agencies, and hospital managers (n = 18). We utilized three focus group discussions with hospital staff and TB doctors and nurses. The χ2 tests and Mann-Whitney U tests were used to analyse quantitative data, and the thematic analysis using a framework approach was applied to analyse qualitative data. RESULTS: Dantu and Yangzhong counties enacted TB-specific case-based payment method in 2014. Jurong County maintained global budget payment but raised the reimbursement rate for TB care. Compared to the baseline, the percentage of TB patients in Dantu and Yangzhong with eight or above outpatient visits increased from 7.5 to 55.1% and from 22.1 to 53.1% in the final survey, respectively. Jurong experienced the opposite trend, decreasing from 63.0 to 9.8%. In the final survey, the total outpatient expenses per patient during a full treatment course in Dantu (RMB 2939.7) and Yangzhong (RMB 2520.6) were significantly higher than those in the baseline (RMB 690.4 and RMB 1001.5, respectively), while the total outpatient expenses in Jurong decreased significantly (RMB 1976.0 in the baseline and RMB 660.8 in the final survey). Health insurance agencies in Dantu and Yangzhong did not approve the original design with outpatient and inpatient expenses packaged together, revealed by qualitative interviews. Furthermore, staff at designated hospitals misunderstood that health insurance agencies would only reimburse actual expenses. Many TB doctors complained about their reduced salary, which might be due to decreased hospital revenue generated from TB care after the payment method reform. CONCLUSIONS: The intended effect on cost containment of case-based payment was not evident in Dantu and Yangzhong. In Jurong, where the global budget payment system maintained with the reimbursement rate enhanced, we found an effect on cost containment, but the quality of TB care might be compromised. The TB-specific case-based payment method could be redesigned to combine payment on outpatient and inpatient expenses and to set an appropriate payment standard for TB care during a full treatment course. Local health insurance agencies have to provide explicit explanations on the payment method. TB care providers should be provided with proper incentives. Monitoring and evluaiton on the quality of TB care should be undertaken at regular intervals.

[Clinicopathological characteristics and prognosis of uterine malignant mixed Mullerian tumor].

OBJECTIVE: To evaluate the clinicopathological characteristics, treatment and prognosis of uterine malignant mixed mullerian tumor. METHODS: The clinical, pathologic and follow-up data of 16 patients with uterine malignant mixed Mullerian tumor treated in our hospital between March, 2003 and June, 2015 were analyzed. RESULTS: The 16 patients had a median age of 58 years at diagnosis, and 13 of them were postmenopausal. The number of patients with FIGO stage Ia, Ib, II, IIIa, IIIc2, and IV was 7, 3, 1, 3, 1, and 1, respectively. In 15 patients who received uterine segment diagnostic curettage, pathological examination all reported malignant results. Among the 15 patients having serum CA125 level test upon admission, 2 had elevated CA125 levels. The overall and disease-free survival rates of the 16 patients were 75% and 68.8%, respectively, and the 3-year survival rate of 13 patients who were followed up for at least 3 years was 72.7%. Two out of 12 patients receiving retroperitoneal lymph?node?dissection?had had postoperative recurrence, as compared with 3 out 4 who did not had the operation; tumor recurrence was found in 3 out of 13 patients receiving postoperative chemotherapy, as compared with 2 out of 3 patients who did not have chemotherapy; tumor recurrence occurred in 1 out of 10 patients receiving radiotherapy, as compared with 4 out of 6 patients without radiotherapy. The recurrence rates in 11 patients with FIGO stage I-II was 18.2%, and that among the 5 patients with FIGO stage III-IV was 60.0%. CONCLUSIONS: Uterine segment diagnostic curettage has a high diagnostic value for uterine malignant mixed Mullerian tumor. FIGO stage is the important prognostic factor for these patients, and early?diagnosis, accurate surgical staging, platinum-based chemotherapy and postoperative pelvic radiotherapy are all associated with a better prognosis.

Genetic polymorphisms in estrogen-related genes and the risk of breast cancer among Han Chinese women.

Exposure to high levels of estrogen is considered an important risk factor for susceptibility to breast cancer. Common polymorphisms in genes that affect estrogen levels may be associated with breast cancer risk, but no comprehensive study has been performed among Han Chinese women. In the present study, 32 single-nucleotide polymorphisms (SNPs) in estrogen-related genes were genotyped using the MassARRAY IPLEX platform in 1076 Han Chinese women. Genotypic and allelic frequencies were compared between case and control groups. Unconditional logistic regression was used to assess the effects of SNPs on breast cancer risk. Associations were also evaluated for breast cancer subtypes stratified by estrogen receptor (ER) and progesterone receptor (PR) status. Case-control analysis showed a significant relation between heterozygous genotypes of rs700519 and rs2069522 and breast cancer risk (OR = 0.723, 95% CI = 0.541-0.965, p = 0.028 and OR = 1.500, 95% CI = 1.078-2.087, p = 0.016, respectively). Subgroup comparisons revealed that rs2446405 and rs17268974 were related to ER status, and rs130021 was associated with PR status. Our findings suggest that rs700519 and rs2069522 are associated with susceptibility to breast cancer among the Han Chinese population and have a cumulative effect with three other identified SNPs. Further genetic and functional studies are needed to identify additional SNPs, and to elucidate the underlying molecular mechanisms.

[Tetramethoxystilbene, a selective CYP1B1 inhibitor, suppresses adipogenesis of C3H10T1/2 pluripotent stem cells].

OBJECTIVE: To investigate the inhibitory effects of tetramethoxystilbene, a selective CYP1B1 inhibitor, on adipogenic differentiation of C3H10T1/2 multi-potent mesenchymal cells. METHODS: In vitro cultured C3H10T1/2 cells at full confluence were induced by adipogenic agents (10 µg/ml insulin, 2 µmol/L dexamethasone and 0.5 mmol/L 3-isobutyl-1-methylxanthine) and exposed simultaneously to TMS at the final concentrations of 1.0, 2.0 or 4.0 µg/ml. Oil Red-O staining was used to observe the cell differentiation. The expression of peroxisome proliferator-activated receptor gamma (PPARγ) and its target genes cluster of differentiation 36 (CD36) and fatty acid binding protein 4 (FABP4) were quantified by real-time RT-PCR and Western blotting. RESULTS: Oil Red-O staining and TG contents revealed that TMS suppressed induced differentiation of C3H10T1/2 cells. TMS exposure of the cells dose-dependently decreased both mRNA and protein expressions of PPARγ, a key nuclear transcription factor during adipogenesis, and also lowered the mRNA expressions of PPARγ target genes CD36 and FABP4. CONCLUSION: TMS can suppress adipogenic differentiation of C3H10T1/2 cells by inhibiting PPARγ

[Umbilicus metastasis in patients with epithelial ovarian cancer : clinical features of 21 patients].

OBJECTIVE: To analyze the clinical features, treatments and prognosis of patients with Sister Mary Joseph's nodule of umbilicus (SMJN) from epithelial ovarian cancer (EOC) patients. METHODS: Among a total of 2642 pathologically diagnosed EOC cases, 21 cases with SMJN were histopathologically diagnosed and had an age range of 40-66 years at Sun Yat-sen University Cancer Center between January 1991 and January 2011. Their clinical data were retrospectively analyzed. RESULTS: The incidence of SMJN in EOC was 0.79 %. The 1, 2 and 5-year survival rates were 61.8%, 26.8% and 9.5% respectively. The diagnosis was confirmed via local excision biopsy, fine-needle aspiration biopsy or gross pathological diagnosis. Univariate analysis showed that patients with progressive disease or relapsing with umbilical metastasis after treatment had worse prognosis than those diagnosed at pre-treatment (22 vs 6 months, P < 0.01) . Patients with suboptimal cytoreductive surgery and/or less than 6 circles of chemotherapy or palliative treatment had worse prognosis than those with optional cytoreductive surgery during 6-8 circles of chemotherapy (21 vs. 4 months). Multivariate analysis showed that the time to diagnose and treatment regimen were independent predictors of survival (relative risk = 41.28, P < 0.01). CONCLUSIONS: SMJN is a rare manifestation of EOC. Improving the diagnostic vigilance, optimal debulking surgery plus regular chemotherapy and other new individualized postoperative treatments may arrest the progression of EOC and prolong patient survival.

Neurogenic locus notch homolog protein 4 and brain-derived neurotrophic factor variants combined effect on schizophrenia susceptibility.

OBJECTIVES: To investigate the relationships between single-nucleotide polymorphisms (SNPs) in NOTCH4 and brain-derived neurotrophic factor (BDNF) with schizophrenia among Han Chinese in Southern China. METHODS: Two NOTCH4 SNPs (rs520688 and rs415929) and two BDNF SNPs (rs2030324 and rs12273539) were examined in 464 schizophrenics and 464 healthy controls from Hunan province in South China, using the Sequenom MassARRAY® iPLEX System. RESULTS: In the study population, rs520688 and rs2030324 were significantly associated with schizophrenia. A decreased risk of schizophrenia was associated with the rs520688 GA genotype (p = 0.035), whereas an increased risk of schizophrenia was associated with the rs2030324 CC/CT genotype (p = 0.044). The genotype distributions of rs415929 in NOTCH4 and rs12273539 in BDNF did not differ significantly between the case and control groups. Although no allele-allele interactions were detected between rs520688 and rs2030324, recombination analysis revealed a combined effect of the two on the susceptibility to schizophrenia, with GA-TT decreasing and CT/CC-GG/GA increasing the risk of schizophrenia. CONCLUSION: In conclusion, rs520688 in NOTCH4 and rs2030324 in BDNF are significantly associated with schizophrenia among Han Chinese in Southern China. The two had a combined effect on the susceptibility to schizophrenia among Han Chinese in Southern China, but this may not be caused by an allele-allele interaction.

A clinicopathological aspect of primary small-cell carcinoma of the uterine cervix: a single-centre study of 25 cases.

AIMS: Small-cell carcinoma is a variant of poorly differentiated neuroendocrine carcinoma. Primary small-cell carcinoma of the cervix (SCCC) is recognised as a rare and aggressive malignant tumour with poor prognosis. In this study, the authors report 25 Chinese cases of SCCC, with a particular focus on their clinical and pathological characteristics. MATERIAL AND METHODS: The records of 25 patients from 4075 Chinese patients with cervical cancer were collected and reviewed, including the patients' age, initial symptoms, cervical tumour size, International Federation of Gynaecology and Obstetrics clinical stage, lymph-node metastasis, treatments and follow-up results. Immunohistochemical detection was performed for cytokeratin, epithelial membrane antigen, neuron-specific enolase (NSE), synaptophysin (Syn), chromogranin A (CgA), neuronal cell adhesion molecules (CD56), thyroid transcriptional factor-1 and S100 protein (S100). RESULTS: The median age of 25 patients with SCCC was 43.7 years. The most common symptom was abnormal vaginal bleeding. Histologically, there were 19 'homogenous' SCCC samples and six samples of SCCC mixed with adenocarcinoma. The proportion of SCCC samples with positive immunoreactivity were 100.0% for NSE, 96.0% for Syn, 68.0% for CD56, 76.0% for CgA, 40.0% for thyroid transcriptional factor-1, 84.0% for epithelial membrane antigen, 68.0% for cytokeratin and 8.0% for S100, respectively. Every patient received one to three types of treatments, including surgery, chemotherapy and radiotherapy. The median survival time of patients was 20.9 months after diagnosis. CONCLUSION: The higher proportion of positive labelling of Syn, CD56, CgA, and NSE in SCCC implicated that they are valuably applied in a differential diagnosis of the malignancy. The patients with SCCC receive one to three types of therapies, including surgery, chemotherapy and radiotherapy, and have a poor prognosis.

[Clinical analysis of 12 cases of ovarian carcinosarcoma].

OBJECTIVE: To evaluate the clinical pathological characteristics, treatment and prognosis of ovarian carcinosarcoma. METHODS: The clinical, pathological and follow-up data of 12 cases of ovarian carcinosarcoma treated in Cancer Center of Sun Yat-sen University from May, 2002 to May, 2009 were analyzed retrospectively. RESULTS: The 12 patients with ovarian carcinosarcoma had a median age of 55 years at diagnosis, among whom 10 were postmenopausal women. The patients sought medical attention for such symptoms as pelvic and/or abdominal pain, abdominal distention and ascites. Ten patients showed elevated serum CA125 level ad admission, and postoperative chemotherapy resulted in lowered CA125 level within normal range in 7 of them; in 8 cases, CA125 level increased with disease recurrence. Pelvic mass was found by such imaging examinations as CT, MRI and ultrasound in all cases. A definite diagnosis was obtained by postoperative pathological examination. All the patients received surgical resection and platinum-based adjuvant chemotherapy. Two patients achieved disease-free survival after the treatment. Disease relapse occurred in 10 cases within 2 years after surgery, among whom 2 showed disease remission after a secondary surgery and/or chemotherapy, and 1 was receiving chemotherapy; death occurred in 5 cases, and 2 cases were lost to the follow-up. CONCLUSIONS: Ovarian carcinosarcoma has a poor prognosis. Primary surgery and platinum-based postoperative adjuvant chemotherapy is the main treatment for ovarian carcinosarcoma. The prognosis of ovarian carcinosarcoma is associated with the residual disease after surgery. The patients with disease recurrence may obtain remission and survival through a secondary surgery and/or chemotherapy. Serum CA125 can be used as a marker for monitoring the chemotherapeutic effect in clinical observation and follow-up visits.

[Effects of pH and Cu2+ on the detection of the Microcystin-LR based on fluorescence immunoassay and their elimination].

The effects of pH and Cu2+ on the detection of the Microcystin-LR based on evanescent wave all-fiber immunosensor (EWAI) and their elimination were studied. The experimental results indicated that both low pH and high pH greatly affected the immunoassay results of MC-LR. When pH was less than 6 or higher than 8, the fluorescence signals detected by immunosensor system obviously reduced with the decrease or increase of pH. While pH ranged from 6 to 8, IC50 and the linear working range of MC-LR calculated from the calibration curve were 1.01-1.04 microg/L and 0.12-10.5 microg/L, respectively, which was favourable for MC-LR immunoassay. Low concentration Cu2+ rarely affected the detection performance of MC-LR. When the concentration of CuSO4 was higher than 5 mg/L, the fluorescence signal detected by EWAI clearly decreased. Furthermore, when the concentration of CuSO4 equalled to 10 mg/L, the fluorescence signal detected reduced 70%. The influence of Cu2+ on the immunoassay could effectively be compromised when 1% EDTA was added to the pre-reaction mixture.

Matrix Effects on the Microcystin-LR Fluorescent Immunoassay Based on Optical Biosensor.

Matrix effects on the microcystin-LR fluorescent immunoassay based on the evanescent wave all-fiber immunosensor (EWAI) and their elimination methods were studied. The results indicated that PBS and humic acid did not affect the monitoring of samples under the investigated conditions. When the pH was less than 6 or higher than 8, the fluorescence signals detected by immunosensor systems were obviously reduced with the decrease or increase of pH. When the pH ranged from 6 to 8, IC(50) and the linear working range of MC-LR calculated from the detection curves were 1.01∼1.04 µg/L and 0.12∼10.5 µg/L, respectively, which was favourable for an MC-LR immunoassay. Low concentrations of Cu(2+) rarely affected the detection performance of MC-LR. When the concentration of CuSO(4) was higher than 5 mg/L, the fluorescence signal detected by EWAI clearly decreased, and when the concentration of CuSO(4) was 10 mg/L, the fluorescence signal detected was reduced by 70%. The influence of Cu(2+) on the immunoassay could effectively be compromised when chelating reagent EDTA was added to the pre-reaction mixture.

[Progress of evanescent wave fluorescence immunosensors in environmental detection].

The evanescent wave fluorescence immunosensor, which employs evanescent wave produced in interface of the waveguide when the light transmits on the manner of total internal reflection in the waveguide, is relatively new kind of sensor on the basis of fluorescence immunoassay principle. This sensor has many advantages including strong specificity, high sensitivity, rapid detection, cost-effective and simple operation, which will be widely applied for environmental detection, clinical medicine and food industry. The principle, structure and characteristics of evanescent wave fluorescence immunosensors were covered and the applications of this kind of sensor in environmental detection were introduced.

[Influence of magnetic field on membrane flux and CaCO3 crystallization in the unstirred dead-end NF process].

With reference to a control system operating under same conditions, unstirred dead-end NF processes of magnetic pretreated solution containing Ca2+ were conducted with NF and NF270 nanofiltration membranes to study the influence of magnetic field on the membrane flux and the crystallization of CaCO3 on membrane surface. Ca2+ in the solution was 3.6 mmol/L, and the hardness was equal to the alkalinity. When experimental solution with initial volume of 300 mL was concentrated 2 times, the flux of membranes treating general solution presents a monotonously falling trend, while the flux of membranes treating magnetic solution decreased firstly and then increased, and then failed again. The average flux in treating magnetic solution was higher than that of the general solution, which indicated that magnetic pretreatment could ameliorate parts of membrane performance. More crystals were formed on membranes treating magnetic solution according to the SEM observation and the chemical analysis. It was assumed that magnetization could speed crystallization on membrane surface, which resulted in concentration falling in the polarization layer and flux increasing accordingly. The formed crystals that may decrease effective membrane areas were all deposited on the membrane surface, so the flux decreased finally. Average flux was higher while more crystals were formed on membranes, which conflicted to the conventional concept of membrane fouling and could not be explained by the serial-resistance model, implied the crystallization on membrane surface unnecessarily detered membrane performance.