[The theory of unresponsive pulse by Wang Ji : The historical position of his Yun Qi Yi Lan].

Wang Ji (1463-1539) was a well-known doctor of the Xin An Medical School in the Ming Dynasty. He and his representative masterpiece Yun Qi Yi Lan are particularly important in the medical history of Yunqi, which refers to the principles of Air (Qi) regulation, influencing almost all life in nature. In terms of the theory "nonresponsive pulse matching the South and the North in the ten Stem years" (Nan Bei Zheng Bu Ying Mai), Wang Ji differentiated and analysed the changes of this theory after the Jin and Yuan Dynasties and traced it back to the classics the Inner canon of Huangdi (Huang Di Nei Jing), based on Su Wen Ru Shi Yun Qi Lun Ao, Huang Di Nei Jing and other relevant reference materials. This paper examined the evolution of the theory of unresponsive pulse in the ancient and modern literature. It was found that after the Song Dynasty, the theory of nonresponsive pulse in the South-North in the ten Stem years was developed into two main schools. One was represented by Cheng Wuji and Liu Wansu, followed with Zhang Jingyue, Li Yanshi, Yao Zhian, Lu Guanquan, Wu Qian, Huang Yuanyu, Xue Fuchen and Zhou Xuehai, who argued that the nonresponsive pulse was determined by the position of Shaoyin. Another was represented by Liu Wenshu, followed with Wang Ji, Li Zhongzi, Zhang Zhicong and Ren Yingqiu, who believed that Shaoyin always stands in the middle, Jueyin and Taiyin are always on the two sides of Shaoyin.

[Role of personalized 3D printing in brain protection after decompressive craniectomy].

Objective: To explore the application value of personalized three-dimensional (3D) printed protective cap in brain protection after decompressive craniectomy (DC). Methods: Fourty-five patients who underwent DC from January 2021 to October 2021 were selected, including 26 males and 19 females, aged 5-73 (50±13) years old. The brain CT data were imported into 3D Slicer software to rebuild the protective cap through 3D printing. The cap was worn on the head of the patient, thereby preventing secondary braindamage. The follow-up results were compared with 53 patients without protective capduring the same period. Results: There were no statistically significant differences in age, skull defect location and follow-up time between the two groups (all P>0.05).Among 45 patients, 47 brain protective caps (2 cases with bilateral skull defects) were successfully designed. The time for image post-processingand 3D printing was (21.2±6.0) min and (62.4±8.3) min, respectively. There were 6 cases of low compliance, 9 cases of moderate compliance, 32 cases of high compliance, respectively. Six cases with low conformity were redesigned and printed, 2 of 9 cases with moderate conformity were redesigned and printed, and the remaining 7 cases reached high compliance after grinding and packaging. In the current study, 45 patients with brain protective caps were followed up for 3 months, and no secondary brain injury occurred. However, among 53 patients without brain protective caps during the same period, 4 patients had secondary accidental brain compression. The incidence of injury was 7.5 %, and the difference was statistically significant (P<0.001). Conclusion: Brain protective cap designed based on cranial CT and 3D printing can be used in patients with skull defects to protect the brain tissue from secondary crush damage and has certain clinical value.

[The mediating role of anxiety and depression for family members of ICU patients in perceived social support and fatigue].

To analyze the mediating role of anxiety and depression in perceived social support and fatigue in ICU patients' families, and to provide a theoretical evidence for alleviating their fatigue status. A total of 223 family members of ICU patients who received treatment at the Affiliated Hospital of Jiangnan University from October 2020 to April 2021 were selected as the study subjects. The general data questionnaire, perceived social support scale (PSSS), generalized anxiety disorder scale (GAD-7), patient health questionnaire (PHQ-9) and fatigue assessment instrument (FAI) were used to conduct a survey. Among 223 family members of ICU patients, 155(69.51%) had fatigue problems. There were statistically significant differences in total fatigue scores of ICU patients' family members in terms of gender, age, education level, relationship with patients, residence, payment method and per capita monthly income (P<0.05). Anxiety, depression and fatigue were negatively correlated with perceived social support (r are -0.353, -0.276 and -0.416, respectively, all P<0.01). Depression and fatigue were positively correlated with anxiety (r are 0.808 and 0.703, respectively, all P<0.01), and fatigue was also positively correlated with depression (r= 0.665, P<0.01). Anxiety and depression had a partial mediating effect on perceived social support and fatigue, and the total indirect effect size was 52.64%. Comprehensive intervention on the level of social support, anxiety and depression is helpful to improve the fatigue status of ICU patients' family members.

[Predictive value of complement and coagulation indicators in sepsis related acute kidney injury].

Objective: To explore the predictive value of complement and coagulation indicators in sepsis related acute kidney injury (AKI). Methods: Clinical data of 217 patients with sepsis admitted to the Department of Internal Medicine and Intensive Care Unit of Affiliated Hospital of Jiangnan University from January 2018 to June 2019 were retrospectively analyzed. All patients were divided into sepsis with AKI group and without AKI group. Laboratory indicators of all patients were collected, including complement C3, complement C4, activated partial thrombin time (APTT), prothrombin time (PT), international normalized ratio (INR), D-dimer, procalcitonin(PCT), etc. logistic regression analysis was used to explore the risk factors of sepsis related AKI. Receiver operating characteristic curve (ROC) was used to evaluate the predictive value of independent risk factors. Results: Among 217 patients, 120 patients developed sepsis related AKI and 97 patients didn't. PCT, lactic acid, PT, APTT, INR and D-dimer in AKI patients were significantly higher than those without AKI (P<0.01). Complement C3 and complement C4 were significantly lower in AKI group (P<0.01). Multivariate logistic regression analysis suggested that blood pressure<90/60 mmHg (1 mmHg=0.133 kPa)(OR=3.705, 95%CI 1.536-8.934,P=0.004), increased lactic acid (OR=1.479, 95%CI 1.089-2.008, P=0.012), decreased complement C3 (OR=0.027, 95%CI 0.005-0.152, P<0.001) and prolonged APTT (OR=1.090, 95%CI 1.047-1.137,P<0.001)were independent risk factors predicting AKI. The area under the ROC curve (AUC) of these multivariates were 0.741 (95%CI 0.675-0.807), 0.798 (95%CI 0.732-0.864), 0.712 (95%CI 0.643-0.781) and 0.716 (95%CI 0.648-0.783) respectively. The relevant sensitivity was 57.5%, 80.8%, 87.5%, 59.2%, and the specificity was 90.7%, 75.3%, 51.5%, 77.3%, respectively. The AUC of the combined four indicators was 0.880 (95%CI 0.835-0.926) with the sensitivity 75.0% and the specificity 90.7%. Conclusion: The low level of complement C3 and prolonged APTT predict sepsis related AKI, and the predictive value can be enhanced if hypotension and hyperlactacidemia are added.

Photosynthetic characteristics of summer maize under different planting patterns and the responses to nitrogen application of previous crop / Características fotossintéticas do milho verão sob diferentes padrões de plantio e as respostas à aplicação de nitrogênio da cultura anterior

Maize (Zea mays L.) is one of the most important grain crops in the North China Plain. Management practices affect the photosynthetic characteristics and the production of summer maize. This two-year (2014-2015) study examined the effects of different planting patterns and the application of nitrogen to previous winter wheat (Triticum aestivum L.) on the photosynthetic characteristics, yield and radiation use efficiency (RUE) of summer maize. Field experiments used a two-factor split-plot design with three replicates at Taian, Shandong Province, China (36°09' N, 117°09' E). The experiments involved two planting patterns(ridge planting, RP; and uniform row planting, UR) and two nitrogen application levels of previous winter wheat (N1, 112.50 kg ha-1; N2, 225.00 kg ha-1). The results indicated that the application of nitrogen on previous crop and ridge planting of the following crop had significant effects on the photosynthetic characteristics and yields of summer maize. Compared with UR, this study found that RP increased the chlorophyll content index (CCI), leaf area index (LAI), net photosynthetic rate (Pn), dry matter (DM), yield and grain RUE by 4.1%, 6.3%, 5.2%, 6.4%, 8.9% and 9.4%, respectively. The CCI, LAI, Pn, yield, and grain RUE of N2 were 9.7%, 3.3%, 3.7%, 10.0% and 10.1% higher than those of N1, respectively. RP combined with the application of nitrogen on previous crop of winter wheat could increase the CCI, LAI, Pn, DM, ultimately increasing the grain yield and RUE of the following summer's maize. It was concluded that previous crop nitrogen application and RP pattern treatment resulted in optimal cropping conditions for the North China plain.

O milho (Zea mays L.) é uma das culturas de grãos mais importantes da Planície do Norte da China. Práticas de manejo afetam as características fotossintéticas e a produção do milho verão. Este estudo de dois anos (2014-2015) examinou os efeitos de diferentes padrões de plantio e a aplicação de nitrogênio ao trigo de inverno anterior (Triticum aestivum L.) sobre as características fotossintéticas, produtividade e eficiência de uso de radiação (RUE) do milho verão. Experimentos de campo usaram um delineamento em parcelas subdivididas de dois fatores com três repetições em Taian, província de Shandong, China (36°09' N, 117°09' E). Os experimentos envolveram dois padrões de plantio (ridge planting, RP; e uniform row planting, UR) e dois níveis de aplicação de nitrogênio do trigo de inverno anterior (N1, 112,50 kg ha-1; N2, 225,00 kg ha-1). Os resultados indicaram que a aplicação de nitrogênio na cultura anterior e no plantio RP da cultura seguinte teve efeitos significativos nas características fotossintéticas e na produtividade do milho verão. Comparado com o plantio UR, este estudo concluiu que RP aumentou o índice de conteúdo de clorofila (CCI), índice de área foliar (LAI), taxa fotossintética líquida (Pn), matéria seca (DM), produtividade e RUE de grãos em 4,1%, 6,4%, 5,2%, 6,4%, 8,9% e 9,4%, respectivamente. Os valores de CCI, LAI, Pn, produtividade e RUE de N2 foram 9,7%, 3,3%, 3,7%, 10,0% e 10,1% superiores aos de N1, respectivamente. RP combinada com a aplicação de nitrogênio na safra anterior de trigo de inverno poderia aumentar os valores de CCI, LAI, Pn, DM, aumentando o rendimento de grãos e RUE do milho do verão seguinte. Concluiu-se que a aplicação prévia de nitrogênio na colheita e o tratamento com padrão RP resultaram em condições ótimas de cultivo para a planície do norte da China.

The effect of ventilator mask atomization inhalation of ipratropium bromide and budesonide suspension liquid in the treatment of COPD in acute exacerbation period on circulating levels of inflammation and prognosis.

OBJECTIVE: We investigated the effects of ventilator mask atomization inhalation of ipratropium bromide and budesonide suspension liquid in the treatment of acute exacerbation COPD (AECOPD) on circulating levels of inflammatory factors and prognosis. PATIENTS AND METHODS: A total of 86 cases of patients on ventilator support were randomly divided into control group and observation group with 43 cases each. The control group was administered routine treatment including basic disease treatment, anti-infection, maintenance of a stable internal environment, nutritional support, oxygen inhalation and so on. The control group was administered saline through a ventilator mask. The observation group was treated with atomized inhalation of ipratropium bromide and budesonide suspension and oxygen flow 3-5 L/min, 15-20 min/time and twice a day for 1 week. The treatment effects were compared. RESULTS: Serum TNF-α, IL-6, and CRP levels were decreased in both groups after treatment, but levels in the observation group were significantly lower than those of the control group; differences were statistically significant (p < 0.05). Forced vital capacity (FVC), forced expiratory volume (FEV1), FEV1/FVC and maximal expiratory flow rate in the observation group were significantly higher than those in the control group after treatment (p < 0.05). After treatment, the PaO2, SpO2 and respiratory failure index (RFI) of the observation group were significantly higher than those of the control group. The PaCO2 levels of the observation group were lower than those of the control group. The differences were statistically significant (p < 0.05). The clinical efficacy of the observation group was better than that of the control group; the ventilation time and total treatment time was significantly shorter and the differences were statistically significant (p < 0.05). CONCLUSIONS: The ventilator mask atomizing inhalation of ipratropium bromide and budesonide suspension liquid in the treatment of AECOPD can significantly improve circulating inflammatory reaction, improve lung function and blood gas levels, increase the treatment efficiency, and shorten the treatment time.

[Arthroscopic treatment of eldly with massive rotator cuff tear].

Objective: To investigate the surgical technique and clinical efficacy of arthroscopic treatment of the elderly patients with massive rotator cuff tear. Methods: From June 2012 to June 2015, thirty-six patients with massive rotator cuff tear were treated with arthroscopic and followed up. The visual analog scale(VAS)pain score, University of California Los Angeles (UCLA) scores, Constant scores and American Shoulder and Elbow Surgeons scale(ASES)were used before and after the arthroscopic surgery. Results: All the patients were followed up for average of 18.5 (12 to 30) months.Before arthroscopic surgery, the VAS, UCLA, Constant, ASES were (6.1±2.2), (10.6±4.3), (40.3±10.5) and (28.8±18.5) points; the average flexion of the shoulder was (76.5±42.6)°, the average abduction of the shoulder was (72.4±35.2)°, the average external rotation of the shoulder was(26.6±22.2)° and the average internal rotation of the shoulder was (20.2±6.2)° respectively.These scores were improved to (1.4±1.2), (30.4±5.2), (82.6±12.6), and (78.8±22.6) points, the average flexion of the shoulder was improved to (152.8±25.6)°, the average abduction of the shoulder was improved to (120.6±32.8)°, the average external rotation of the shoulder was improved to (42.6±16.2)° and the average internal rotation of the shoulder was improved to (38.4±5.6)° after one-year follow-up period.Improvement in these scores and range of motion(ROM) were significant difference(P<0.05). Conclusion: Arthroscopic repair can effectively treat the eldly patient with massive rotator cuff tear and obviously improve the function of shoulder joint. The surgery has a clinical application value.

Morphological and phylogenetic identification of Botrytis sinoviticola, a novel cryptic species causing gray mold disease of table grapes (Vitis vinifera) in China.

Seventy-five isolates of Botrytis collected from table grapes (Vitis vinifera) with gray mold symptoms in China were identified based on morpho-cultural characteristics on potato dextrose agar (20 C) and/or phylogenetic analysis using the sequences of three nuclear genes (G3PDH, HSP60, RPB2). Isolates of different species of Botrytis were compared with fenhexamid sensitivity, Bc-hch gene-RFLP haplotyping and pathogenicity to V. vinifera. The 75 isolates comprise two species, B. cinerea (63 isolates) and an undescribed Botrytis sp. (12 isolates) described here as Botrytis sinoviticola Zhang et al. sp., nov. Both B. sinoviticola (Bs) and B. cinerea (Bc) were found to have 20 C optimum for mycelial growth and 25 C for conidial germination. Sensitivity to fenhexamid was significantly greater (P < 0.05) for Bc (EC50 = 0.04 ± 0.01 µg mL(-1)) than for Bs (EC50 = 0.08 ± 0.02 µg mL(-1)). Digestion of the PCR amplicons of the Bc-hch gene with Hha I generated two haplotypes, Group I haplotype for Bs and Group II haplotype for Bc. Bs infected table grapes (leaves, berries) only through wounds, whereas Bc infected both injured and non-injured tissues of table grapes. This study suggests that Bs is a cryptic species sympatric with Bc on table grapes in China.

A novel multi-parameter scoring system for distinguishing sarcoidosis from sputum negative tuberculosis.

BACKGROUND: Sarcoidosis is a granulomatous disorder of unknown cause, affecting multiple organs. Tuberculosis is the world's second most common cause of death from infectious diseases. Due to the similar clinical, radiological and histopathological pictures in sarcoidosis and tuberculosis, Mycobacterium tuberculosis has been considered as potential infectious factor. However, it remains difficult to distinguish sarcoidosis from tuberculosis, especially when sputum examinations for mycobacterium are negative. METHODS: 1. to establish a scoring system for differentiating sarcoidosis and tuberculosis: We collected the risk factors, laboratory data and the data of clinical, radiographic, pathological manifestations from the 117 of sarcoidosis patients and 181 of sputum negative tuberculosis patient. And we put them into the designed form. Based on the results of univariate analysis, clinical experience and the literature, we further selected 13 variables that were more supportive to distinguish the two diseases. Finally 9 variables were selected based on logistic regression to establish the scoring systems with significant differences between the two diseases. The beta-coefficient form the logistic regression were used to calculate the weight of each variable. Four types of comprehensive scoring models were established in the end (clinical-- radiographic; clinical--radiographic--radionuclide; clinical--radiographic--pathological and clinico-radiographic--radionuclide--pathological group). Receiver operating characteristics (ROC) analysis was used to determine an optimal cutoff point for each scoring system. 2. to validate the accuracy of the established scoring system: 73 of new sarcoidosis patients and 57 of new tuberculosis patients were chosen to assess the diagnosis accuracy of the four scoring systems. RESULTS: 1. we established four types of comprehensive scoring models, included clinical--radiographic; clinical--radiographic--radionuclide; clinical--radiographic--pathological and clinico--radiographic--radionuclide--pathological scoring models, the optimal cutoff values respectively were 9, 17, 18 and 22, the sensitivity and specificity of the four scoring system to distinguish the two diseases respectively were: 93.16% (109/117) and 97.79% (177/181), 92.31% (108/117) and 98.90% (179/181); 93.16% (109/117) and 98.90% (179/181); 94.87% (111/117) and 98.90% (179/181). 2. Validation of the scoring systems with 130 new patients (73 of sarcoidosis and 57 of tuberculosis):, the sensitivity and specificity of CR, CRE, CRP, CREP were 91.78% (67/73) and 87.72% (50/57), 97.26% (69/73) and 98.25% (56/57), 94.52% (71/73) and 96.49% (55/57), 98.63% (72/73) and 98.25% (56/57) respectively. CONCLUSIONS: The four scoring systems established by this study can be utilized to differentiate sarcoidosis and sputum negative tuberculosis effectively. Based on the availability of clinical-radiographical/histopathological data, any of the four diagnostic scoring systems were reliable tools for differential diagnosis, with increased information leading to better discrimination.

Control of postharvest Botrytis fruit rot of strawberry by volatile organic compounds of Candida intermedia.

A study was conducted to identify volatile organic compounds or volatiles produced by Candida intermedia strain C410 using gas chromatography-mass spectrometry, and to determine efficacy of the volatiles of C. intermedia in suppression of conidial germination and mycelial growth of Botrytis cinerea and control of Botrytis fruit rot of strawberry. Results showed that, among 49 volatiles (esters, alcohols, alkenes, alkanes, alkynes, organic acids, ketones, and aldehydes) identified from C. intermedia cultures on yeast extract peptone dextrose agar, two compounds, 1,3,5,7-cyclooctatetraene and 3-methyl-1-butanol, were the most abundant. Synthetic chemicals of 1,3,5,7-cyclooctatetraene; 3-methyl-1-butanol; 2-nonanone; pentanoic acid, 4-methyl-, ethyl ester; 3-methyl-1-butanol, acetate; acetic acid, pentyl ester; and hexanoic acid, ethyl ester were highly inhibitory to conidial germination and mycelial growth of B. cinerea. Inhibition of conidial germination and mycelial growth of B. cinerea by volatiles of C. intermedia was also observed. Meanwhile, results showed that incidence and severity of Botrytis fruit rot of strawberry was significantly (P < 0.01) reduced by exposure of the strawberry fruit to the volatiles from C. intermedia cultures or C. intermedia-infested strawberry fruit. These results suggest that the volatiles of C. intermedia C410 are promising biofumigants for control of Botrytis fruit rot of strawberry.

A major SNP resource for dissection of phenotypic and genetic variation in Pacific white shrimp (Litopenaeus vannamei).

Bioinformatics and re-sequencing approaches were used for the discovery of sequence polymorphisms in Litopenaeus vannamei. A total of 1221 putative single nucleotide polymorphisms (SNPs) were identified in a pool of individuals from various commercial populations. A set of 211 SNPs were selected for further molecular validation and 88% showed variation in 637 samples representing three commercial breeding lines. An association analysis was performed between these markers and several traits of economic importance for shrimp producers including resistance to three major viral diseases. A small number of SNPs showed associations with test weekly gain, grow-out survival and resistance to Taura Syndrome Virus. Very low levels of linkage disequilibrium were revealed between most SNP pairs, with only 11% of SNPs showing an r(2)-value above 0.10 with at least one other SNP. Comparison of allele frequencies showed small changes over three generations of the breeding programme in one of the commercial breeding populations. This unique SNP resource has the potential to catalyse future studies of genetic dissection of complex traits, tracing relationships in breeding programmes, and monitoring genetic diversity in commercial and wild populations of L. vannamei.

First Report of Garlic Leaf Blight Caused by Botrytis porri in China.

In the spring of each year from 2007 to 2009, a leaf blight of garlic (Allium sativum L.) was observed in more than 50 fields in Zhushan County of Hubei Province, China. Gray mold was observed on many of the blighted garlic leaves. The percentage of garlic plants with blight and gray mold symptoms ranged from 10 to 50% with one to three blighted leaves on each plant, which severely reduced the yield of young garlic plants (produced as a green vegetable). Ten strains of a Botrytis sp. were isolated from symptomatic garlic leaves collected from 10 different fields. These strains were inoculated onto potato dextrose agar (PDA) in petri dishes and incubated at 20°C for 3 to 15 days for observation of colony characteristics and morphology of sclerotia and conidia. All 10 Botrytis strains formed flat and "ropy" mycelia (mycelial strands) on PDA. Abundant sporulation with a gray powdery appearance was observed on the colonies after 6 days. Conidiophores were erect with alternate branches at the top and ranged from 907 to 1,256 µm high. Conidia were borne in botryose clusters on conidiophores, obovate, and 10.4 to 17.6 × 7.6 to 13.1 µm with an average length/width ratio of 1.36. Discrete sclerotia were produced on each colony after 15 days. Mature sclerotia were black, cerebriform and convoluted, and 1.9 to 9.1 × 1.6 to 6.5 mm. Morphological characteristics of the colonies, conidia, and sclerotia of these Botrytis strains were similar to Botrytis porri Buchwald (1,2). Strain GarlicBC-16 was selected as a representative for molecular identification. Genomic DNA was extracted from mycelia of this strain and used as a template for amplification of the internal transcribed spacer (ITS) region of rDNA using primer pair ITS1/ITS4. A 539-bp amplicon was obtained and sequenced (GenBank Accession No. EU519206). Excluding the flanking regions, the amplicon contained a 453-bp ITS sequence (ITS1 + 5.8S rDNA + ITS2) 100% identical to the ITS sequence of strain MUCL3234 of B. porri (GenBank Accession No. AJ716292). Pathogenicity of strain GarlicBC-16 was tested by inoculation of 10 young and fully expanded garlic leaves taken from 100-day-old garlic plants with mycelial agar plugs (three plugs per leaf and spaced by 5 cm). Ten garlic leaves inoculated with agar plugs of PDA alone served as controls. Inoculated garlic leaves were covered with a plastic film (0.1 mm thick; Gold Mine Plastic Industrial Ltd. Jiangmen, China) and incubated at 20°C with 12-h light/12-h dark. Control leaves remained healthy after 48 to 120 h, but gray, water-soaked lesions appeared on leaves inoculated with strain GarlicBC-16 after 48 h. The average lesion length reached 27.3 mm after 90 h and abundant sporulation was produced on necrotic leaf lesions after 120 h. Microscopic examination showed the shape and size of conidia that formed on garlic leaf lesions were similar to those formed by strain GarlicBC-16 on PDA. On the basis of the isolation, identification, and pathogenicity tests, B. porri was determined to be the causal agent of garlic leaf blight in Zhushan County. B. porri has been reported to cause neck rot of leek (A. porrum) (1) and clove rot of garlic (2), and has been isolated from asymptomatic foliage and seeds of A. cepa (3). To our knowledge, this is the first report of garlic leaf blight caused by B. porri in China. References: (1) S. K. Asiedu et al. Plant Dis. 70:259, 1986. (2) F. M. Dugan et al. J. Phytopathol. 155:437. 2007. (3) L. J. du Toit et al. Plant Dis. 86:1178, 2002.

First Report of Pestalotiopsis microspora Causing Leaf Blight of Reineckea carnea in Central China.

Pink reineckia (Reineckea carnea (Andrews) Kunth) is an evergreen herbaceous perennial plant widely grown as groundcover or for medical purposes in southern China. In 2006 and 2007, severe leaf blight was observed on pink reineckia in Wuhan, China. On newly formed pink reineckia leaves, symptoms were first noted in early May as grayish to dark brown, oval or irregular-shaped lesions, 1.5 to 0.2 × 0.5 to 0.1 cm (n = 50), on the leaf margin or leaf tip. A yellowish halo surrounded each lesion. Lesions enlarged and coalesced and diseased leaves became blighted during the fall and winter. In severely infected plots, most plants became straw-colored and had to be replaced with healthy seedlings. A fungus was isolated from surface-disinfested lesions on potato dextrose agar (PDA) at a frequency of 85.7%. One of 30 isolates, designated C2, was characterized further. The fungus growing on PDA at 20°C for 14 days formed zonate white colonies and black acervular conidiomata. Conidia of the fungus aggregated on acervuli as droplets. Conidia were fusiform and 20.7 to 32.2 × 5.8 to 9.8 µm (n = 50). Each conidium had one hyaline apical cell, one hyaline basal cell, and three dark brown median cells. There were two to four hyaline filamentous appendages 8.1 to 20.4 µm long attached to each apical cell and one hyaline appendage 2.4 to 7.1 µm long attached to each basal cell. The cultural and morphological characteristics of isolate C2 matched the description for Pestalotiopsis microspora (Speg.) Batista & Peres (1,2). The internal transcribed spacer (ITS) region of the ribosomal DNA (ITS1-5.8S-ITS2) was PCR-amplified and sequenced. The ITS sequence (606 bp) for isolate C2 (GenBank Accession No. EU935587) was 100% similar to P. microspora isolates TA-57 (GenBank Accession No. AY924267) and LK32 (GenBank Accession No. DQ001002). Pathogenicity of isolate C2 was tested with the method described by Keith et al. (2). Four detached leaves were wound inoculated or inoculated without wounding with mycelia on agar plugs (4 mm in diameter; three plugs per leaf) or conidial suspensions (107 conidia per ml; 20 µl on each of three sites per leaf). Control leaves were wound inoculated with PDA or sterile water. All inoculated leaves were maintained in a moist enamel tray under fluorescent light for 7 days at 20°C. The test was performed twice. After 4 days of incubation, necrotic leaf lesions resembling symptoms that occurred in the field were observed on the wound-inoculated leaves, whereas the control leaves and C2-inoculated leaves without wounding remained healthy. Therefore, wounding was necessary for symptom development (2). A fungus was reisolated from the C2-induced leaf lesions and the morphology of colonies and conidia were identical to that for isolate C2 of P. microspora. On the basis of the results of isolations, inoculations, and fungal identification, P. microspora was determined to be the causal agent for leaf blight of pink reineckia occurring in Wuhan, China. This fungus previously has been reported as the causal agent of scab disease of Psidium guajava in Hawaii (2), decline of Torreya taxifolia in Florida (3), and leaf blight of Lindera obtusiloba in Korea (1). To our knowledge, this is the first report of the occurrence of P. microspora on R. carnea. References: (1) Y. H. Jeon et al. Plant Pathol. 56:349, 2007. (2) L. M. Keith et al. Plant Dis. 90:16, 2006. (3) M. W. Schwartz et al. Plant Dis. 80:600, 1996.

Expression of Foxp3+CD4+CD25+ regulatory T cells and Th1/Th2, Tc1/Tc2 profiles in the peripheral blood of patients with condyloma acuminatum.

BACKGROUND: Condyloma acuminatum (CA) is a disease that appears as proliferative lesions of the genital epithelium caused by human papillomavirus (HPV) infection. The balance between type 1 and type 2 T-cell subsets in patients with CA is thought to modulate antiviral immunity. CD4+CD25+ regulatory T cells (Tregs) inhibit proliferation and cytokine production by both T-helper (Th)1 and Th2 cells and reversibly suppress CTL-mediated immunity. A better understanding of the mechanisms of T-cell regulation in CA might help in developing more effective therapeutic strategies. Objective. To evaluate the balance of Th1/Th2 and Tc1/Tc2 and to assess their correlation with changes in number of Tregs in CA. METHODS: The percentage of Th1, Th2, Tc1, Tc2 and Tregs were detected by flow cytometry after intracellular staining for cytokines (interferon-gamma and interleukin-4) and Foxp3 of T lymphocytes in the peripheral blood of 30 patients and 20 healthy volunteers. RESULTS: Patients with CA showed a decreased proportion of Th1 and Tc1 cells and a decreased ratio of Th1/Th2 and Tc1/Tc2. In particular, strikingly decreased ratios of Th1/Th2 were found in 15 patients with relapsed CA (P < 0.01). The mean +/- SD number of Foxp3+CD4+CD25+ Tregs increased significantly in patients with CA (3.37 +/- 1.03%) and patients with relapsed CA (4.68 +/- 1.17%) compared with healthy controls (1.18 +/- 0.53%) (P < 0.001). CONCLUSION: Tregs appear to downregulate cytokine expression in both Tc1 and Th1 subsets of effector T cells, which may be responsible for antivirus responses.

First Report of Onion Bulb Rot Caused by Botrytis aclada in China.

During the spring of 2006, onion bulbs with gray mold symptoms on the surface were observed in a few supermarkets in Wuhan, China. Onions mummified as they decayed. Further surveys of five randomly selected batches of onion bulbs in one of the supermarkets indicated that the disease occurred in all batches and the disease incidence ranged from 6 to 50%. Eight diseased onion bulbs were collected arbitrarily and isolations were made using homemade potato dextrose agar (PDA). Single-spore cultures of the isolated Botrytis sp. were established and maintained on PDA plates at 20°C. The 10-day-old PDA cultures of all of these isolates were gray and covered with abundant beige, ovoid- or oblong-shaped conidia, which were budded from terminal ampullae formed on dichotomously branching conidiophores. Conidia from these isolates measured 7.6 to 10.4 µm long and 4.2 to 5.6 µm wide, with an average of 8.4 × 5.0 µm. No sclerotia were produced from any of these PDA cultures after incubation at 20°C for 30 days. Morphological characteristics of colonies and conidia of these isolates were similar to Botrytis aclada according to the description made by Yohalem et al. (3). Inoculation of healthy onion bulbs with one of the eight fungal strains, OnionBc-15, resulted in gray mold symptoms similar to those observed in the supermarkets. Microscopic examinations showed that the size and shape of conidia that formed on the surface of diseased bulbs of onion were identical to the size and shape of conidia of OnionBc-15, indicating that this isolate can cause onion bulb rot. The isolate OnionBc-15 was further characterized by molecular techniques. Genomic DNA was extracted from mycelia of this strain and used as a template for amplification of two previously reported DNA regions, the internal transcribed spacer (ITS) region of the ribosomal RNA genes and the L45-550 sequence (1), which can be used to distinguish B. aclada and two closely related species, B. allii and B. byssoidea (3). Universal primers ITS1 and ITS4 were used to amplify the ITS region (2). A 539-bp DNA sequence was generated, cloned, and sequenced (GenBank Accession No. EU093077). The sequence contained two SphI restriction sites and was 99% identical in nucleotides to that of B. aclada strain PRI006 (GenBank Accession No. AJ716295). It is different from B. allii and B. byssoidea, which have only one SphI restriction site for the ITS1/ITS4-amplified DNA sequence (2). The Botrytis-specific primers, BA2f and BA1r, were used to amplify the L45-550 sequence (2). A 413-bp DNA sequence was generated, cloned, and sequenced. The sequence did not contain any ApoI restriction sites. This is also similar to B. aclada, but different from B. allii and B. byssoidea, which contains one ApoI restriction site in the BA2f/BA1r-amplified DNA sequence (2,3). On the basis of morphological characteristics and the two molecular features, it is concluded that the isolate OnionBc-15 belongs to B. aclada. To our knowledge, this is the first report on the occurrence of B. aclada causing onion bulb rot in China. References: (1) K. Nielsen and D. S. Yohalem. Mycologia 93:264, 2001. (2) K. Nielsen et al. Plant Dis. 86:682, 2002. (3) D. S. Yohalem et al. Mycotaxon 85:175, 2003.

Hypovirulence and Double-Stranded RNA in Botrytis cinerea.

ABSTRACT Twenty-one strains of Botrytis cinerea isolated from 13 species of plants grown in China were compared for pathogenicity on Brassica napus, mycelial growth on potato dextrose agar, and presence of double-stranded (ds)RNA. The results showed that the strain CanBc-1 was severely debilitated in pathogenicity and mycelial growth, compared with the 20 virulent strains. A dsRNA of approximately 3.0 kb in length was detected in CanBc-1 and 4 hypovirulent single-conidium (SC) isolates of CanBc-1, but was not detected in the 20 virulent strains of B. cinerea and 4 virulent SC isolates of CanBc-1. Results of the horizontal transmission experiment showed that the hypovirulent trait of CanBc-1 was transmissible and the 3.0-kb dsRNA was involved in the transmission of hypovirulence. Analysis of a 920-bp cDNA sequence generated from the 3.0-kb dsRNA of CanBc-1 indicated that the dsRNA element was a mycovirus, designated as B. cinerea debilitation-related virus (BcDRV). Further analyses showed that BcDRV is closely related to Ophiostoma mitovirus 3b infecting O. novo-ulmi, the causal agent of Dutch elm disease. Mitochondria and cytoplasm in hyphal cells of CanBc-1 became degenerated, compared with the virulent isolate CanBc-1c-66 of B cinerea. This is the first report on the occurrence of Mitovirus-associated hypovirulence in B. cinerea.

Construction and characterization of a cDNA library from 4-week-old human embryo.

Development and differentiation studies of early human embryos have been severely impeded by general difficulties in obtaining suitable samples. In order to isolate and identify new genes expressed during early human development, we constructed and characterized a PCR-based cDNA library using a 4-week-old chorion-free human embryo. The constructed cDNA library contained 6.3 x 10(6) directional recombinants, and its insert size ranged from 0.4 to 1.8 kb. The cDNA library proportionally represents the mRNA population, containing beta-actin, tPA and LINE1 repetitive sequences at the expected frequencies as in other conventionally constructed and PCR-based cDNA libraries. PCR analyses of the library for specific genes have also revealed the presence of cDNAs for developmentally important genes such as CD59, MCP, Quox-1 and ZNF268. Among the 70 randomly selected cDNA clones, 53% encoded previously known genes, 26% matched with anonymous sequences, and 17% showed no sequence similarity and were designated as human early embryo-specific ESTs. These results demonstrate the sequence complexity and relatively low redundancy of our cDNA library. Furthermore, approximately 40% of those randomly analyzed clones contained full-length encoding regions. To our knowledge, this is the first description of the PCR-based cDNA library from a 4-week-old chorion-free human embryo, and the presence of novel sequences within this library makes it a valuable and unique resource for studying gene expression and regulatory mechanisms that underlie the early process of human embryogenesis.

Cloning and characterization of a novel Krüppel-like zinc finger gene, ZNF268, expressed in early human embryo.

With the aim of identifying genes involved in early human embryonic development, we have isolated a cDNA clone representing a novel human zinc finger gene ZNF268 from 3 week old human embryo cDNA library using a differential hybridization strategy. The complete cDNA sequence of ZNF268 contained an open reading frame of 2841 nucleotides that encodes a 947 amino acid protein with an N-terminal Krüppel-associated box (KRAB) domain and 24 C-terminal zinc fingers. Northern blot analysis showed that ZNF268 mRNA is mainly expressed in 3-5 week old human embryos suggesting it could play certain roles in the embryogenesis. The gene consists of six exons spanning about 22 kb of genomic DNA. According to the genomic sequence from the HTGS database, the ZNF268 gene is assigned to human chromosome 5.