[Analysis of clinical features of cervical precancerous lesions in postmenopausal women].

Objective: To analyze the characteristics of high-grade squamous intraepithelial lesion (HSIL) diagnosed by cervical tissue sampling in postmenopausal women. Methods: A retrospective study was performed on 2 013 patients with HSIL diagnosed by cervical tissue sampling under colposcopy and treated by cervical conization at the First Affiliated Hospital of Zhengzhou University from June 2017 to November 2018, to compare the difference of patients' clinical features, HPV test, liquid-based thin-layer cytology (TCT), performance of colposcopy and biopsy pathology, pathology after cervical conization between 439 postmenopausal patients and 1 574 pre-menopausal patients. Results: (1) Clinical features: the proportion of contact bleeding showed no significant difference between postmenopausal patients and pre-menopausal patients [4.3% (19/439) vs 6.4% (101/1 574); χ²=2.672, P=0.102]. Among the patients with contact bleeding, the proportion of cervical cancer after cervical cone resection was significantly higher in postmenopausal patients compared with pre-menopausal patients [10/19 vs 22.8% (23/101); χ²=7.157, P=0.007]. Among the patients found by routine screening, the proportion of cervical cancer after cervical cone resection was significantly higher in postmenopausal patients compared with pre-menopausal patients [9.0% (38/420) vs 4.3% (63/1 473); χ²=14.726, P<0.01]. The proportion of smooth cervix was higher in postmenopausal patients compared with pre-menopausal patients [63.6% (279/439) vs 35.5% (558/1 574); χ²=111.601, P<0.01]. (2) High-risk HPV infection: there was no significant difference in the high-risk HPV positive rate between the postmenopausal group and the pre-menopausal group [92.0% (404/439) vs 94.4% (1 486/1 574); χ²=3.394, P=0.065]; the HPV 16 infection was the most common type, but there was no significant difference in the HPV 16 infection rate between the two groups [65.8% (289/439) vs 68.0% (1 070/1 574); χ²=0.722, P=0.395]. (3) TCT test: TCT test results included negative for intraepithelial lesion and malignancy (NILM), atypical squamous cell of undetermined signification (ASCUS), atypical squamous cells cannot exclude high-grade lesion (ASC-H), low grade squamous intraepithelial lesion (LSIL), HSIL, compared with the different results of TCT examination, there were not statistically significant difference between postmenopausal and pre-menopausal patients (all P>0.05). (4) The performance of colposcopy: the proportion of insufficient colposcopy and the proportion of cervical type Ⅲ conversion area were higher in postmenopausal patients compared with pre-menopausal patients [87.5% (384/439) vs 32.5% (511/1 574), P<0.01; 80.0% (351/439) vs 21.9% (344/1 574), P<0.01]. The proportion and positive rate of endocervical curettage (ECC) in postmenopausal patients were higher than those in pre-menopausal patients [35.3% (155/439) vs 20.4% (322/1 574), P<0.01; 67.7% (105/155) vs 53.1% (171/322), P=0.003]. The proportion of lesions involving the vaginal wall was higher in postmenopausal patients compared with pre-menopausal patients [5.9% (26/439) vs 1.0% (16/1 574); χ²=40.443, P<0.01]. There was a positive correlation between vaginal wall lesions and cervical lesions in postmenopausal patients (r=0.660, P<0.01). (5) Postoperative pathology: the positive rate of margin and the proportion of pathological escalation after cervical conization were significantly higher in postmenopausal patients compared with pre-menopausal patients [14.6% (64/439) vs 4.8% (75/1 574), 10.9% (48/439) vs 5.5% (86/1 574); P<0.01]. Conclusions: Colposcopy in postmenopausal women is often inadequate, and the cervix is mostly type Ⅲ transformation zone. The lesion in postmenopausal women is more likely to involve the cervical canal and vaginal wall. Clinical attention should be paid to cervical tube curettage and comprehensive examination of the vaginal wall. The high rate of positive margins and a high proportion of pathological upgrading after cervical conization in postmenopausal patients requires further active intervention.

[Grading evaluation of operative complications and analysis of related risk factors in patients with stage â endometrial cancer treated by robotic-assisted and traditional laparoscopic surgery].

Objective: To investigate the surgical complications in the treatment of stage Ⅰ endometrial cancer by robotic-assisted laparoscopy, the risk degree of Clavein-Dindo complications and the main risk factors affecting the occurrence of surgical complications. Methods: A retrospective case-control study was conducted in the First Affiliated Hospital of Zhengzhou University from October 2014 to June 2019. The patients were divided into robotic-assisted laparoscopy group and traditional laparoscopy group according to the operation mode, including 131 cases in robot group and 290 cases in traditional laparoscopy group. To compare the complications during and after operation and the risk degree of complications between the two groups by Clavein-Dindo classification standard, the age, body mass index (BMI), comorbidities, past history of pelvic surgery, American Society of Anesthesiologists (ASA) grade, preoperative anemia, number of pelvic lymph node resection, number of abdominal aortic lymph node resection, the total number of lymph node resection, operation time, surgical methods (robot surgery or traditional laparoscopic surgery) and other clinicopathological data were analyzed by logistic regression analysis. Results: (1) Complications of operation: the incidence of operative complications (including intraoperative and postoperative complications) in robot group was significantly lower than that in traditional laparoscopy group [(20.6%, 27/131) vs (34.8%, 101/290); χ(2)=8.620, P=0.003)]. The incidence of intraoperative complications in robot group was lower than that in traditional laparoscopy group [1.5% (2/131) vs 6.2% (18/290); χ(2)=4.368, P=0.037]. The incidence of intraoperative vascular injury in robot group was significantly lower than that in traditional laparoscopy group [0.8% (1/131) vs 5.2% (15/290); χ(2)=4.798, P=0.022]. The incidence of postoperative complications in robot group was also lower than that in traditional laparoscopy group [19.1% (25/131) vs 28.6% (83/290); χ(2)=4.303, P=0.038], but the incidence of postoperative lymphatic leakage in robot group was higher than that in traditional laparoscopy group [10.7% (14/131) vs 5.2% (15/290); χ(2)=4.279, P=0.039]. (2) Clavein-Dindo classification: the incidence of Clavein-Dindo Ⅰ, Ⅲ, Ⅲ, Ⅳ and Ⅴ grade between two groups were respectively 3.8% (5/131) vs 11.0% (32/290), 13.7% (18/131) vs 14.5% (42/290), 3.1% (4/131) vs 8.6% (25/290), 0 (0/131) vs 0.3% (1/290), 0 (0/131) vs 0.3% (1/290), and the incidence of grade Ⅰ (χ(2)=5.684, P=0.015) and Ⅲ (χ(2)=4.361, P=0.037) complications were statistically significant. The incidence of severe complications in robot group (grade Ⅲ and above) was lower than that in traditional laparoscopy group [3.1% (4/131) vs 9.3% (27/290); χ(2)=5.179, P=0.023]. (3) Analysis of influencing factors of surgical complications: univariate analysis showed that BMI (χ(2)=15.801, P=0.000), preoperative anemia (χ(2)=14.299, P=0.000), total number of lymph node resection (χ(2)=10.425, P=0.001), surgical methods (χ(2)=8.620, P=0.003) were related to the occurrence of surgical complications of endometrial carcinoma. Multivariate analysis showed that BMI (OR=0.289, 95%CI: 0.097-0.864, P=0.026), preoperative anemia (OR=0.309, 95%CI: 0.129-0.740, P=0.008), the total number of lymph node resection (OR=0.624, 95%CI: 0.403-0.966, P=0.034) and surgical methods (OR=3.491, 95%CI: 1.030-11.840, P=0.045) were independent risk factors for surgical complications of endometrial carcinoma. Conclusions: Compared with traditional laparoscopic surgery, robot-assisted laparoscopic surgery has fewer complications and lower incidence of severe complications. BMI, preoperative anemia, the total number of lymph node resection and surgical methods are independent risk factors for the occurrence of surgical complications of stage Ⅰ endometrial cancer.

miR-375 inhibits the proliferation and invasion of glioblastoma by regulating Wnt5a.

The aberrant expression of microRNA-375 (miR-375) has been proved to be associated with carcinogenesis. However, the role of miR-375 in glioblastoma (GBM) remains unknown. The aim of this study was to investigate biological functions and its molecular mechanisms of miR-375 in GBM cells. In this study, real-time PCR results showed that the level of miR-375 expression in GBM tissues and GBM cell lines (U87 and U251) was decreased. Using MTT assay, Transwell migration and invasion assay, we demonstrated that miR-375 overexpression significantly suppress cell proliferation, cell migration and cell invasion capacity in U87 and U251 cells. However, downregulation of miR-375 had reverse effects on cell proliferation, migration and invasion. Targeting association analysis, dual luciferase assay, RT-PCR and western blot analysis results confirmed that miR-375 could target the 3'UTR of Wnt5a mRNA and regulated its protein expression. Further studies also find overexpression of Wnt5a could significantly reverse miR-375-mediated proliferation, migration and invasion on U87 and U251 cells. Therefore, we concluded that miR-375 inhibited the proliferation and invasion of GBM by regulating Wnt5a and might be a possible therapeutic agent for GBM.

[A multicenter, randomized, controlled, phase â £ clinical study of PEG-rhG-CSF for preventing chemotherapy induced neutropenia in patients with breast cancer].

Objective: To explore the efficacy and safety of polyethylene glycal recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) in preventing chemotherapy-induced neutropenia in patiens with breast cancer. Methods: There were two parts in the present phase Ⅳ clinical study. One was a randomized, controlled clinical study. Patients in this study received PEG-rhG-CSF or rhG-CSF in the first cycle and followed with both PEG-rhG-CSF in the rest of 3 cycles. The other one was a single arm study. Patients who developed Ⅲ/Ⅳ grade neutropenia in the screening cycle received PEG-rhG-CSF in the rest of 3 cycles chemotherapy. Results: In the first cycle of randomized, controlled study, the incidence of Ⅳ grade neutropenia are 31.48% and 35.58% respectively in PEG-rhG-CSF and rhG-CSF group, with no statistically significant differences (P=0.527 6). The duration of Ⅳ grade neutropenia respectively are 2.22±1.58 and 3.00±1.59 days, with a statistically significant difference (P=0.016 6). In the single arm study, the incidence of Ⅳ grade neutropenia was 57.76% in screening cycle. And the incidence decreased to 16.35%, 10%, and 8.57% in the followed 3 cycle after the use of PEG-rhG-CSF. The incidence of adverse effects was 5.06%, and the major adverse effect was bone pain which with an incidence of 2.8%. Conclusion: The fixed 6mg dose of PEG-rhG-CSF can effectively prevent neutropenia in patients with breast cancer in multicycle chemotherapy and it has a low incidence of adverse events and mild adverse reaction.

Clinical efficacy and mechanism of lymphoplasma exchange in the treatment of Guillain-Barre syndrome.

Guillain-Barre syndrome (GBS) is an autoimmune disease of the nervous system and is the most common acute polyneuropathy. Both cellular and humoral immunity are believed to be involved in the pathogenesis of GBS, and various types of activated CD4+ T cells are thought to orchestrate the onset and progression of GBS. Lymphoplasma exchange (LPE) filtering out activated lymphocytes while exchanging plasma has been used for GBS treatment for years. However the treatment is still not yet optimal. In order to assess the efficacy of this treatment, we evaluate the effect of LPE and determine the appropriate frequency of LPE treatments for GBS patients through comparing the neurological deficit scores and the changes in related immunology indicators of GBS patients before and after LPE treatment. Twenty-four patients with GBS who received LPE were evaluated for immunologic indicants before treatment, on the second day, and the fourth day after the treatment. The immunoglobulin complement and CD4+ T lymphocyte subsets were tested by flow cytometry. The patients' Medical Research Council sum scores were increased from 25.7±10.4 up to. 36.7±10.4 (P=0.019) and their Hughes scores decreased from 3.7±0.76 to 3.1±0.73 (P=0.027) at 7 days after LPE. In the peripheral blood from patients received LPE treatment, the levels of immunoglobulin, complement, monocytes and fibrinogen were significantly reduced. The percentages of Th1 and Th17 cells in the CD4+ T lymphocyte subsets were significantly decreased, whereas the Th2 and Treg cells were increased in patients after treatment. The changes in CD4+T lymphocyte subsets were correlated with patient MRC score changes. Our data indicate that LPE is effective in treating GBS patients by directly removing immunoglobulin, complement, monocytes, and fibrinogen as well as regulating lymphocyte subsets in the peripheral blood.

Evaluation of a loop-mediated isothermal amplification assay based on hrpZ gene for rapid detection and identification of Pseudomonas syringae pv. lachrymans in cucumber leaves.

AIMS: Cucumber angular leaf spot caused by Pseudomonas syringae pv. lachrymans (Psl) is an important and destructive disease worldwide, and no effective technique has been developed for the control of the pathogen. Detection of infection or latent in cucumber plants is critical to evaluate disease progress and strengthening management to avoid a serious epidemic in the fields. In this paper, we developed a rapid and sensitive method for detection of Psl using an isothermal method known as loop-mediated amplification (LAMP). METHODS AND RESULTS: A set of six primers was designed to amplify the gene coding for the hrpZ, and conditions for detection were optimized to complete in 60 min at 67°C, and the amplification were confirmed through gel electrophoresis or visually inspected using calcein stain. The specificity of LAMP primers set was widely validated on Psl and nontarget strains. In sensitivity testing, LAMP allowed detection as low as 104  CFU per ml bacterial cells without DNA extraction. The novel method was also applied for detecting Psl in infected cucumber leaves, and even the early onset of disease can be detected by the assay. CONCLUSIONS: This study confirmed that the novel developed LAMP assay is an easy, rapid and sensitive method for the detection of Psl in infected leaves. SIGNIFICANCE AND IMPACT OF THE STUDY: The method is suitable for direct detection of Psl without strain enrichment and complex DNA extraction from samples in the field, and hence it has the capability to be used for on-site disease diagnosis and field surveys.

[Telbivudine for prevention of perinatal transmission in pregnant women infected with hepatitis B virus in immune-tolerant phase: a study of efficacy and safety of drug withdrawal].

OBJECTIVE: To observe the success rate of telbivudine (LdT) for the prevention of perinatal transmission of hepatitis B virus (HBV) and the incidence of alanine aminotransferase (ALT) elevation during LdT treatment and after LdT withdrawal in HBV-infected pregnant woman with high viremia in immune-tolerant phase and receiving LdT treatment at the end of pregnancy, and to evaluate the efficacy of LdT in the prevention of perinatal transmission and the safety for pregnant women. METHODS: Pregnant women infected with HBV in immune-tolerant phase who had normal ALT levels (≤40 U/L) and high viremia (HBV DNA ≥6 log10 IU/ml) with positive HBeAg were enrolled as subjects. All pregnant women received antiviral treatment with LdT at the end of pregnancy to prevent perinatal transmission of HBV. All infants received standard combined immunoprophylaxis. Failure for prevention of perinatal transmission of HBV was defined as positive HBsAg or HBV DNA in infants 7 months of age (or at one month after the third injection of hepatitis B vaccine). Liver function, HBV DNA, and HBV serological markers were evaluated at baseline, after 1 month of treatment, before childbirth, and 1, 3, and 6 months after drug withdrawal. SPSS 16.0 software was used to analyze the data. Between-group comparison of continuous data was made by t test, and comparison of categorical data was made by chi-square test. RESULTS: One hundred and four pregnant women (treatment group) received oral administration of 600 mg LdT once a day, and 25 pregnant women (observation group) did not receive any antiviral therapy. The success rate for the prevention of perinatal transmission was significantly higher in the treatment group than in the observation group (100% vs 89.47%, χ (2) = 9.862, P = 0.028). There was no significant difference in the incidence of ALT elevation during treatment and within 6 months after drug withdrawal between the treatment group and the observation group (4.81% (5/104) vs 4.00% (1/25), χ (2) = 0.030, P = 1.000). In the treatment group, the mean HBV DNA at baseline was significantly higher than that before childbirth (8.20±0.78 vs 3.98±0.90 log10IU/ml, t = 6.979, P < 0.001). One hundred patients with drug withdrawal had HBV DNA increased to 8.11±0.80 log10 IU/ml at one month after childbirth. CONCLUSION: LdT treatment at the end of pregnancy can effectively reduce the incidence of perinatal transmission of HBV in pregnant women with high viremia in immune-tolerant phase. The immediate drug withdrawal after childbirth is safe for the mother. The incidence of hepatitis is low after drug withdrawal.

TBX21 polymorphisms are associated with virus persistence in hepatitis C virus infection patients from a high-risk Chinese population.

Hepatitis C virus (HCV) infection is a major cause of chronic liver disease and the varied outcomes of the infection depend on both viral and host factors. We have demonstrated that the HCV alternate reading frame protein (F protein) is related to Th1/Th2 bias which is involved in virus persistence in chronic hepatitis C (CHC) patients. The purpose of this study was to test the hypothesis that genetic variants of TBX21 (T cell specific T-box transcription factor) were associated with the outcomes of HCV infection and F protein generation. Three single nucleotide polymorphisms (SNPs) (rs17250932, rs2074190, rs4794067) in the TBX21 gene were genotyped in a case-control study in a cohort of a high-risk group, including 354 healthy controls and 747 CHC patients (190 anti-F protein antibody seronegative patients and 557 anti-F protein antibody seropositive patients). Results showed that the rs4794067 C allele in the TBX21 promoter was significantly more common in CHC patients (OR = 1.335, 95% CI = 1.058-1.684, P = 0.015), exceptionally in anti-F protein seropositive patients (OR = 1.547, 95% CI = 1.140-2.101, P = 0.005), compared with healthy controls. And the risk effect was also significantly high in patients with HCV 1b genotype and mild fibrosis (P = 0.021, P = 0.010, respectively). Compared with the most frequent haplotype TAT, haplotype analysis showed that the distribution of TAC was significantly different between the chronic HCV carrier group and the healthy group, and so was the anti-F antibody seronegativity group and the anti-F antibody seronegativity group (all P < 0.001). Our results suggested that TBX21 variants may be involved in the etiology of this disease.

S-phase kinase-associated protein 2 expression interference inhibits breast cancer cell proliferation.

We investigated the expression of S-phase kinase-associated protein 2 (SKP2) in breast cancer tissues, and the effects of SKP2-specific small interfering RNA (siRNA) interference on breast cancer cell proliferation. Thirty subjects provided breast cancer tissue samples and 18 subjects provided normal breast specimens for this study. The expression of SKP2 in breast cancer patient tissues and normal breast tissues was detected by western blotting analysis and reverse transcription-polymerase chain reaction. SKP2-specific siRNA was used to decrease SKP2 expression in breast cancer cell line MDA-MB-231. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect cell proliferation. SKP2 expression in breast cancer tissues was significantly higher than in normal breast tissues (P < 0.05). Two pairs of siRNA specific to SKP2 were required to downregulate SKP2 expression in the breast cancer cell line MDA-MB-231. The MTT assay showed that MDA-MB-231 growth significantly slowed after SKP2 interference. Patients with breast cancer have an increased SKP2 level. Interference in SKP2 gene expression can inhibit breast cancer cell growth, suggesting that SKP2 is potentially a new target for breast cancer therapy.

Effect of lipoic acid combined with paclitaxel on breast cancer cells.

Breast cancer is the most common gynecologic tumor globally that threatens women's health. Lipoic acid is a type of antioxidant that can alleviate oxidative stress damage. Studies showed that lipoic acid could inhibit the proliferation of tumor cells in cervical cancer and colon cancer. This paper intends to explore the combined effect of lipoic acid and paclitaxel on breast cancer cells. Breast cancer MCF-7 cells were divided into four groups: control group, lipoic acid group, paclitaxel group, and a combination group. MTT was applied to detect the drugs' influence on breast cancer cell proliferation. A colony formation test was used to determine the effects on breast cancer cell clone formation rate. Western blot was performed to detect the effects on nuclear factor (NF)-κB. Lipoic acid alone can inhibit tumor cell proliferation and clone formation with time dependence. Compared with the control, paclitaxel alone can significantly suppress tumor cell proliferation and clone formation (P < 0.05). Lipoic acid and paclitaxel in combination obviously strengthened their individual inhibitory effects on tumor cells (P < 0.05). Compared with the paclitaxel alone group, the combination group exhibited more remarkable inhibitory effect (P < 0.05). Lipoic acid alone or combined with paclitaxel can inhibit NF-κB expression and inhibit breast cancer cell proliferation.

Investigation of ERCC1 and ERCC2 gene polymorphisms and response to chemotherapy and overall survival in osteosarcoma.

We assessed the role of single nucleotide polymorphisms (SNPs) in ERCC1 and ERCC2 genes in the clinical outcomes for osteosarcoma patients receiving cisplatin-based treatment. A perspective study was conducted on 260 patients with osteosarcoma during 2010 and 2011. A polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) assay was used to assess the ERCC1 rs11615 and rs3212986, and the ERCC2 rs1799793 and rs13181 gene polymorphisms. After adjustment for clinical variables, we found that the CC genotype of ERCC1 rs11615 was significantly associated with better response to chemotherapy (OR = 2.87, 95%CI = 1.24-6.97). Our study found that those carrying the CC genotype of ERCC1 rs11615 had a longer overall survival compared with the TT genotype, and the OR (95%CI) was 0.35 (0.12-0.92). In conclusion, our results suggest that the ERCC1 rs11615 polymorphism might influence the response to cisplatin-based chemotherapy and affect the clinical outcome for osteosarcoma patients.

First Report of Colletotrichum capsici Causing Anthracnose on Hosta plantaginea in China.

Fragrant plantain lily [Hosta plantaginea (Lam.) Aschers.] is an easily grown herbaceous perennial plant valued for its decorative foliage and dainty colorful flowers. From 2009 to 2011, a leaf spot disease of H. plantaginea was observed in Yuyuantan Park in Beijing, China (116°25' E, 39°55' N). The leaf spots began as small, irregular, circular, brown lesions in the middle or on the margin of leaves, which enlarged gradually up to 1 to 20 mm in diameter and were circular or irregular and brown to dark brown surrounded by yellowish borders. Occasionally, some spots cracked under dry conditions. Symptomatic leaf tissues were surface-sterilized in 1% NaOCl for 2 min, washed three times with distilled water, and then placed on potato dextrose agar (PDA). Colonies on PDA at 25°C for 7 days were grayish brown and cottony. Mycelia were hyaline to grey, septate, branched, and 2 to 7 µm wide. Acervuli were dark brown to black and 198 to 486 µm in diameter, averaging 278.5 µm. Setae were pale brown to dark brown, 2 to 4 septa, 70.0 to 120.3 × 2.5 to 5.1 µm, base cylindrical, and narrower towards the apex. Conidiophores were unicellular, hyaline, phialidic, and 5.0 to 13.5 × 1.5 to 2.8 µm. Conidia were hyaline, aseptate, falcate, apices acute, oil globules, and 16.0 to 25.2 × 2.6 to 5.0 µm. Appressoria were spherical, ovate or obclavate, pale to dark brown, edge usually entire, and 9.5 to 15.5 × 6.5 to 11.5 µm. Morphological characteristics of the fungus were similar to those of Colletotrichum capsici (Syd.) Butler & Bisby (2). To validate Koch's postulates, pathogenicity tests were performed by spraying leaves of 20 healthy potted H. plantaginea (60-day-old plants) with a 106 conidia/ml aqueous suspension. Control plants were inoculated with sterile water. Plants were put into a glass cabinet for 48 h after inoculation and maintained at 25°C, relative humidity 98%. Then the plants were moved out and incubated in greenhouse at 10 to 25°C. After 10 days, all inoculated plants showed typical symptoms, whereas water sprayed controls remained healthy. C. capsici was consistently re-isolated from these lesions. The re-isolated fungus showed the same morphological characteristics as described above. Genomic DNA was extracted from the original isolate and the re-isolate from the pathogenicity test. PCR amplification of the internal transcribed spacer (ITS) regions from ribosomal DNA was performed with primers ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3'). PCR products of 513 bp were sequenced. There was 100% nucleotide identity for sequences of the original isolate and the re-isolate. The sequence was submitted to GenBank (Accession No. HM063417.1). BLAST analysis of the fungal sequence resulted in 100% identity to the sequence of C. capsici (Accession No. JX867217.1). Isolates have been deposited at the Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences. To our knowledge, this is the first report of anthracnose caused by C. capsici on H. plantaginea in China (1). Its confirmation is a significant step toward management recommendations for growers. References: (1) D. F. Farr and A. Y. Rossman, Fungal Databases. Syst. Mycol. Microbiol. Lab. ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , August 2013. (2) J. E. M. Mordue. CMI Description of Pathogenic Fungi and Bacteria. Commonwealth Mycol. Inst., Kew, UK, 1971.

First Report of Pseudomonas cichorii Causing Leaf Spot of Vegetable Sponge Gourd in China.

A suspect bacterial leaf spot on vegetable sponge gourd (Luffa cylindrical (L.) Roem.) was found in a commercial greenhouse in Pi County, Chengdu City, Sichuan Province, China, in February 2011. Approximately 20 to 30% of plants were affected, causing serious economic loss. Symptoms occurred only on seedlings and consisted of water-soaked, irregularly shaped, black lesions on the surface and margins of cotyledons. A bacterium was consistently isolated on nutrient agar from diseased leaf tissues that had been surface disinfected in 70% ethyl alcohol for 30 s. The bacterium produced small gray colonies with smooth margins, was gram negative, fluoresced on King's B medium, and showed pectolytic activity when inoculated on potato slices. The partial sequences of 16SrRNA gene (1,377 bp) of the bacterium (GenBank Accession No. KC762217), amplified by using universal PCR primers 16SF (5'-AGAGTTTGATCCTGGCTCAG-3') and 16SR (5'-GGTTACCTTGTTACGACTT-3'), shared 100% similarity with that of Pseudomonas cichorii (GenBank Accession No. HM190228). The vegetable sponge gourd isolate was also identified by using the Biolog Microbial Identification System (version 4.2, Biolog Inc., Hayward, CA) as P. cichorii with the following characteristics (1): negative for arginine dihydrolase, gelatin liquefaction, and N2 production. Positive reactions were obtained in tests for catalase, oxidase, potato rot, utilization of melibiose, and mannitol. Tests were negative for utilization of sucrose, trehalose, D-arabinose, raffinose, cellobiose, and rhamnose. A pathogenicity test was conducted on 4-week-old vegetable sponge gourd plants by spray-inoculation with 108 CFU/ml sterile distilled water on the leaves of 15 vegetable sponge gourd plants and by needle puncture on the stems of 15 other plants with P. cichorii, respectively. Control plants were misted with sterile distilled water or punctured on the stem with a clean needle. Plants were placed in a greenhouse maintained at 28 ± 2°C with relative humidity of 80 to 85%. Symptoms, the same as seen on the original diseased plants, developed after 7 to 10 days on inoculated plants. Control plants remained healthy. The bacterium was readily re-isolated from inoculated plants and identified as P. cichorii using P. cichorii-specific primer hrpla/hrp2a (1). To our knowledge, this is the first report of P. cichorii causing disease on commercially grown vegetable sponge gourd in China. This new finding will provide the basis for developing resources for diagnostics and management, including screening varieties for resistance. References: (1) S. Mazurier et al. J. FEMS Microbiol. Ecol. 49:455, 2004. (2) N. W. Schaad et al., eds. Laboratory Guide for Identification of Plant Pathogenic Bacteria, 3rd ed. APS Press, St. Paul, MN, 2001.

First Report of Corynespora Leaf Spot Caused by Corynespora cassiicola on Gynura in China.

Gynura (Gynura bicolor DC.) is a perennial herbaceous plant in the family Compositae. It is an important Chinese vegetable, and is commonly used as a Chinese herbal medicine. In 2010, a severe leaf spot disease was observed on gynura grown in the main production areas in Tong Nan County, Chongqing City, China. Some farms experienced 60% disease incidence. Symptoms usually began on the lower leaves, as circular to elliptical or irregular spots with concentric rings. Individual spots were dark brown with grayish centers, sometimes coalescing and leading to extensive necrosis. The fungus associated with lesions was characterized as follows: Conidiophores were single or in clusters, straight or flexuous, unbranched, percurrent, cylindrical, pale to dark brown, 87.5 to 375.0 µm long and 5.0 to 10.5 µm wide. Conidia were solitary or catenate, straight to slightly curved, obclavate to cylindrical, 3 to 14 pseudoseptate, 82.8 to 237.5 µm long and 7.0 to 7.8 µm wide, and pale brown. The morphological characteristics of the conidia and conidiophores agreed with the descriptions for Corynespora cassiicola (1). To isolate the causal pathogen, surface-sterilized tissue at the margin of lesions was immersed in 75% ethanol for 30 s, rinsed in sterile water, dried in a laminar flow bench, transferred to PDA, and incubated at 28°C. Four single-spore cultures of the isolates were obtained and named from ZBTK10110637 to ZBTK10110640. All strains were identified as C. cassiicola. The isolate ZBTK10110637 was selected as representative for molecular identification. Genomic DNA was extracted by CTAB (2). The internal transcribed spacer (ITS) region of the rDNA was amplified using primers with ITS1 (5'-TCCGATGGTGAACCTGCGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3'). Amplicons were 433 bp (GenBank Accession No. JX867272) and shared 100% similarity with that of C. cassiicola (NRC2-1 No. AB539285.1). To confirm pathogenicity, four isolates were used to inoculate 12 gynura plants (6 weeks old) by mist spray-inoculation with 108 spores/ml suspension in sterile distilled water on the leaves. Control plants were misted with sterile distilled water. After inoculation, all plants were incubated in a greenhouse maintained at 20 to 28°C with relative humidity of 80 to 85%. Five days after inoculation, dark brown spots with a grayish center typical of field symptoms were observed on all inoculated plants. No symptoms were seen on water-treated control plants. The fungus was re-isolated from inoculated plants. The morphological characteristics of isolates were identical with the pathogen recovered originally. This is the first report of C. cassiicola on gynura. References: (1) M. B. Ellis. CMI Mycological Papers 65(9):1-15, 1957. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

First Report of Myrothecium roridum Causing Leaf Spot on Zantedeschia aethiopica in China.

Zantedeschia aethiopica (L.) Spreng. (calla lily), belonging to family Araceae, is a popular ornamental plant in China. In the summer of 2010, leaves of calla lily with typical symptoms of necrotic lesions were observed in a commercial glasshouse in Beijing, China (116°20' E, 39°44' N). The initial symptoms were circular to subcircular, 1 to 3 mm, and dark brown lesions on the leaf lamina. Under high humidity, lesions expanded rapidly to 5 to 10 mm with distinct concentric zones and produced black sporodochia, especially on the backs of leaves. Later, the infected leaves were developing a combination of leaf lesions, yellowing, and falling off; as a result, the aesthetic value of the plant was significantly impacted. Leaf samples were used in pathogen isolation. Symptomatic leaf tissues were cut into small pieces and surface sterilized with 70% ethanol for 30 s and then in 0.1% mercuric chloride solution for 1 to 3 min. After being washed in sterile distilled water three times, the pieces were plated on potato dextrose agar (PDA) and incubated at 25°C in darkness for 7 days (5). Initial colonies of isolates were white, floccose mycelium and developed dark green to black concentric rings that were sporodochia bearing viscid spore masses after incubating 5 days. Conidiophores branched repeatedly. Conidiogenous cells were hyaline, clavate, and 10.0 to 16.0 × 1.4 to 2.0 µm. Conidia were hyaline, cylindrical, both rounded ends, and 6.0 to 8.2 × 1.9 to 2.4 µm. Morphological characteristics of the fungus were consistent with the description of Myrothecium roridum Tode ex Fr. (3,4). To confirm the pathogenicity, three healthy plants of calla lily were inoculated with a conidial suspension (1 × 106 conidia per ml) brushed from a 7-day-old culture of the fungus. Control plants were sprayed with sterile water. The inoculated plants were individual with clear plastic bags and placed in a glass cabinet at 25°C. After 7 days, all inoculated leaves developed symptoms similar to the original samples, but control plants remained disease free. Re-isolation and identification confirmed Koch's postulates. For molecular identification, genomic DNA of a representative isolate (MTL07081001) was extracted by modified CTAB method (1), and the rDNA-ITS region was amplified by using primers ITS1 (5-TCCGTAGGTGAACCTGCGG-3) and ITS4 (5-TCCTCCGCTTATTGATATGC-3). The 465-bp amplicon (GenBank Accession No. KF761293) was 100% identity to the sequence of M. roridum (JF724158.1) from GenBank. M. roridum has an extensive host range, covering 294 host plants (2). To our knowledge, this is the first record of leaf spot caused by M. roridum on calla lily in China. References: (1) F. M. Ausubel et al. Current Protocols in Molecular Biology. John Wiley & Sons Inc, New York, 1994. (2) D. F. Farr and A. Y. Rossman, Fungal Databases. Syst. Mycol. Microbiol. Lab., ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , October 2013. (3) M. T. Mmbaga et al. Plant Dis. 94:1266, 2010. (4) Y. X. Zhang et al. Plant Dis. 95:1030, 2011. (5) L. Zhu et al. J. Phytopathol. 161:59, 2013.

[Epidemiological characteristics of reinfection of 2019-nCoV and influencing factors in Ningbo].

Objective: To analyze the epidemiological characteristics of reinfection of 2019-nCoV and influencing factors, and provide evidence for effective prevention and control of COVID-19 epidemic. Methods: The incidence data of COVID-19 in Ningbo from January 1, 2020 to November 30, 2022 were collected from the infectious disease surveillance system of Chinese information system for disease control and prevention. The incidence of reinfection of 2019-nCoV was investigated by using questionnaire. logistic regression analysis was used to analyze the influences of gender, age, time interval from the first infection, history of underlying disease, 2019-nCoV vaccination dose and disease severity on the reinfection. Results: A total of 897 previous 2019-nCoV infection cases were investigated, of which 115 experienced the reinfection of 2019-nCoV, the reinfection rate was 12.82%. The interval between the two infections M(Q1, Q3) was 1 052 (504, 1 056) days. Univariate analysis showed that age, 2019-nCoV vaccination dose, history of underlying disease, type of 2019-nCoV variant causing the first infection, time interval from the first infection and severity of the first infection were associated with the reinfection rate (all P<0.05). Multivariate logistic regression analysis showed that the risk for reinfection in age group 30- years was higher than that in age group ≥60 years (OR=2.10, 95%CI: 1.11-3.97). No reinfection occurred in those with time interval from the first infection of <6 months, and the risk for reinfection was higher in those with the time interval of ≥12 months than in those with the time interval of 6- months (OR=6.68, 95%CI: 3.46-12.90). The risk for reinfection was higher in the common or mild cases than in the asymptomatic cases (OR=2.64, 95%CI: 1.18-5.88; OR=2.79, 95%CI: 1.27-6.11). Conclusion: The time interval from the first infection was an important influencing factor for the reinfection of 2019-nCoV, and the probability of the reinfection within 6 months was low.