[Research progress on scleral perforating vessels in the formation of myopic choroidal neovascularization].

Myopic choroidal neovascularization (mCNV) is a severe complication of pathological myopia. Currently, the primary treatment involves anti-vascular endothelial growth factor (VEGF) therapy, which significantly improves visual acuity in mCNV patients. However, challenges such as high recurrence rates and inconsistent therapeutic outcomes persist. Previous studies attributed mCNV to choroidal thinning and ischemia. Recent research suggests that, in addition to choroidal factors, perforating scleral vessels (PSV) are closely associated with the occurrence and therapeutic efficacy of mCNV. This review comprehensively explores the definition of PSVs, their imaging classifications and features, as well as their intricate connections with the occurrence and clinical outcomes of mCNV. By delving into the role and potential mechanisms of PSVs in mCNV, this review aims to deepen our understanding of their involvement in this condition.

[Preliminary observation on the differential expression of metformin in preventing noise-induced hearing loss in inner ear protein group of rats].

Objective: To study the protective effects of metformin on noise-induced hearing loss (NIHL) and its differential protein omics expression profile. Methods: In January 2021, 39 male Wistar rats were randomly divided into control group, noise exposure group and metformin+noise exposure group, with 13 rats in each group. Rats in the noise exposure group and metformin+noise exposure group were continuously exposed to octave noise with sound pressure level of 120 dB (A) and center frequency of 8 kHz for 4 h. Rats in the metformin+noise exposure group were treated with 200 mg/kg/d metformin 3 d before noise exposure for a total of 7 d. Auditory brainstem response (ABR) was used to test the changes of hearing thresholds before noise exposure and 1, 4, 7 d after noise exposure in the right ear of rats in each group. Tandem mass tag (TMT) quantitative proteomics was used to identify and analyze the differentially expressed protein in the inner ear of rats in each group, and it was verified by immunofluorescence staining with frozen sections. Results: The click-ABR thresholds of right ear in the noise exposure group and metformin+noise exposure group were significantly higher than those in the control group 1, 4, 7 d after noise exposure (P<0.05) . The click-ABR threshold of right ear in the metformin+noise exposure group were significantly lower than that in the noise exposure group (P<0.05) . Compared with the noise exposure group, 1035 up-regulated proteins and 1145 down-regulated proteins were differentially expressed in the metformin+noise exposure group. GO enrichment analysis showed that the significantly differentially expressed proteins were mainly involved in binding, molecular function regulation, signal transduction, and other functions. Enrichment analysis of KEGG pathway revealed that the pathways for significant enrichment of differentially expressed proteins included phosphatidylinositol 3-kinase-protein kinase B (PI3K-Akt) signaling pathway, focal adhesion, diabetic cardiomyopathy, mitogen, and mitogen-activated protein kinase (MAPK) signaling pathway. Immunofluorescence experiments showed that compared with the noise exposure group, the fluorescence intensity of insulin-like growth factor 1 receptor (IGF1R) in the metformin+noise exposure group was increased, and the fluorescence intensity of eukaryotic translation initiation factor 4E binding protein 1 (eIF4EBP1) was decreased. Conclusion: Noise exposure can lead to an increase in rat hearing threshold, and metformin can improve noise-induced hearing threshold abnormalities through multiple pathways and biological processes.

Tunable valley polarization in Janus WSSe by magnetic proximity coupling to a CrI3 layer.

We investigate the electronic properties and valley physics of Janus monolayer WSSe on a CrI3 substrate layer based on first-principles calculations. It is shown that the K and K' valley degeneracy can be lifted which leads to valley polarization (VP) in the WSSe due to the magnetic proximity coupling to a magnetic substrate. The magnitude of VP is highly sensitive to the interfacial electronic properties and can be tuned by varying the stacking configurations of the heterostructure. Interestingly, the direction of VP can be altered by manipulating the layer alignment without reversing the magnetism orientation of the magnetic substrate CrI3. We suggest that the hybridization between the bands of WSSe and the substrate plays an important role. Meanwhile, the charge distributions have been mapped out to uncover the microscopic origin of the direction variable VP. In addition, large VP can be achieved by adjusting the interlayer spacing. Our investigations may have potential applications in the design of valleytronic devices.

[The ergonomic analysis of different holding gun methods for standing guard soldiers].

Objective: To study the effects of different holding gun methods and gun weight on health when standing guard, and propose a way to support the health of long-term standing guard soldiers. Methods: We created different percentile mannequins by Classic JACK, and adjusted the standing guard posture based on its standards for soldiers. The pressure on lumbar L4/L5 and moment on ankles and knees were analysied for different holding gun methods and gun weight. Then the mathematical models of joint load, gun weight and body mass index were studied by multiple regression analysis. Results: Holding gun methods and gun weight influence the force characteristics on ankles, knees and lumbar L4/L5. Holding gun with a brace and hands applying downward force -2 kgf could significantly reduce lumbar L4/L5 pressure. When the hand force is -5, -3, -4, -3, -2, -1, 0, 1, 2 kgf, and the weight of the gun is 0, Lumbar vertebrae L4/L5 joint pressure of people with different body mass index(P(1), P(5), P(55), P(95), P(99)) are the smallest, respectively 269, 281, 321, 408, 444 N, and the same change trend occurs when the weight of the gun is 2, 4, and 8 kg.The moment on ankles and knees were less with the same holding gun method and the hands downward force ranged from 0 to -4 kgf, and the higher the body mass index is, the more the hands downward force needed to make the moment on ankles and knees zero. That is, the moment on ankles could be zero when the hands downward force ranged from -1 to -3 kgf, the moment on knees could be zero when the hands downward force ranged from -1.1 to -3.7 kgf. Conclusion: To reduce the pressure on lumbar L4/L5 and moment on ankles and knees, so as to cut down occupational risk of long-standing operation, we advise the long-term standing guard soldiers holding gun with a brace and hands applying downward force -2 kgf.

Effect of Sustained Joint Loading on TMJ Disc Nutrient Environment.

The temporomandibular joint (TMJ) disc nutrient environment profoundly affects cell energy metabolism, proliferation, and biosynthesis. Due to technical challenges of in vivo measurements, the human TMJ disc extracellular nutrient environment under load, which depends on metabolic rates, solute diffusion, and disc morphometry, remains unknown. Therefore, the study objective was to predict the TMJ disc nutrient environment under loading conditions using combined experimental and computational modeling approaches. Specifically, glucose consumption and lactate production rates of porcine TMJ discs were measured under varying tissue culture conditions (n = 40 discs), and mechanical strain-dependent glucose and lactate diffusivities were measured using a custom diffusion chamber (n = 6 discs). TMJ anatomy and loading area were obtained from magnetic resonance imaging of healthy human volunteers (n = 11, male, 30 ± 9 y). Using experimentally determined nutrient metabolic rates, solute diffusivities, TMJ anatomy, and loading areas, subject-specific finite element (FE) models were developed to predict the 3-dimensional nutrient profiles in unloaded and loaded TMJ discs (unloaded, 0% strain, 20% strain). From the FE models, glucose, lactate, and oxygen concentration ranges for unloaded healthy human TMJ discs were 0.6 to 4.0 mM, 0.9 to 5.0 mM, and 0% to 6%, respectively, with steep gradients in the anterior and posterior bands. Sustained mechanical loading significantly reduced nutrient levels (P < 0.001), with a critical zone in which cells may die representing approximately 13.5% of the total disc volume. In conclusion, this study experimentally determined TMJ disc metabolic rates, solute diffusivities, and disc morphometry, and through subject-specific FE modeling, revealed critical interactions between mechanical loading and nutrient supply and metabolism for the in vivo human TMJ disc. The results suggest that TMJ disc homeostasis may be vulnerable to pathological loading (e.g., clenching, bruxism), which impedes nutrient supply. Given difficulties associated with direct in vivo measurements, this study provides a new approach to systematically investigate homeostatic and degenerative mechanisms associated with the TMJ disc.

[The protective effect of Xuebijing on paraquat-induced HK-2 cells apoptosis and the underlying mechanisms].

Objective: To investigate the protective effect and mechanism of Xuebijing (XBJ) on paraquat (PQ) -induced apoptosis in Human kidney cell line-2 (HK-2) cells. Methods: Routinely cultured HK-2 cells, (1) Cell growth inhibition experiment after PQ and XBJ intervention: PQ was divided into 0、200、400、800、1600 and 3200 µmol/L PQ groups, and the cell survival rate was detected after intervening 24、48 and 72 h. XBJ was divided into 0、5、10、20、40 mg/ml XBJ groups, and the cell survival rate was detected after intervening 24、48 and 72 h.To determine the rational drug concentration and the duration of action of XBJ and PQ. (2) PQ-induced HK-2 cell growth inhibition experiment antagonized by XBJ: The cells were divided into normal control group, PQ group (800 µmol/L) and PQ+XBJ group (The cells were pretreated with 5、10 and 20 mg/ml XBJ for 1 h, then cultured with PQ of 800 µmol/L) , After cultured 24 h、48 h and 72 h separately, the cell survival rate was detected. (3) HK-2 cells were divided into normal control group、PQ group (800 µmol/L PQ cultured for 24 h) 、PQ+XBJ group (pretreated with 10 mg/ml XBJ for 1 h, and then 800 µmol/L PQ cultured for 24 h) and XBJ group (10 mg/ml XBJ cultured 24 h). The apoptosis of cells was detected by flow cytometry. The protein expression of Bcl-2 and BAX in each group was detected by Western blotting. The expressions of caspase-3 and caspase-9 were detected by caspase-3 and caspase-9 activity kit active. Results: (1) PQ could significantly reduced the survival rate of HK-2 cells and showed time and concentration dependence. The survival rate of HK-2 cells was about 55% after 800 µmol/L PQ contacted 24 h, XBJ under 20 mg/ml was no significant effect on the survival rate of HK-2 cells after cultured 72 h. (2) Compared with the PQ group, the survival rate of HK-2 cells of PQ+XBJ group was significantly increased (P<0.05). (3) Compared with the normal control group, the cell apoptosis rate of PQ group was significantly increased (P<0.05). Compared with the PQ group, the cell apoptosis rate of PQ+XBJ group was significantly decreased (P<0.05). (4) Compared with the normal control group, Bcl-2 protein expression in PQ group was significantly decreased and BAX protein expression in PQ group was significantly increased (P<0.05) ; compared with PQ group, Bcl-2 protein expression in PQ+XBJ group was significantly increased, BAX protein expression in PQ+XBJ group was significantly decreased (P<0.05). (5) Compared with the normal control group, the activities of caspase-3 and caspase-9 in PQ group were significantly increased (P<0.05). Compared with PQ group, the activities of caspase-3 and caspase-9 in PQ+XBJ group were decreased significantly (P<0.05) . Conclusion: XBJ (10 mg/ml) has obvious protective effect on HK-2 cell injuried by PQ (800 µmol/L) , It can improve the survival rate of cells through reducing the apoptosis of HK-2 cells which induced by PQ.

Fluid pressurization and tractional forces during TMJ disc loading: A biphasic finite element analysis.

OBJECTIVES: To investigate the ploughing mechanism associated with tractional force formation on the temporomandibular joint (TMJ) disc surface. SETTING AND SAMPLE POPULATION: Ten left TMJ discs were harvested from 6- to 8-month-old male Yorkshire pigs. MATERIALS AND METHODS: Confined compression tests characterized mechanical TMJ disc properties, which were incorporated into a biphasic finite element model (FEM). The FEM was established to investigate load carriage within the extracellular matrix (ECM) and the ploughing mechanism during tractional force formation by simulating previous in vitro plough experiments. RESULTS: Biphasic mechanical properties were determined in five TMJ disc regions (average±standard deviation for aggregate modulus: 0.077±0.040 MPa; hydraulic permeability: 0.88±0.37×10-3 mm4 /Ns). FE simulation results demonstrated that interstitial fluid pressurization is a dominant loading support mechanism in the TMJ disc. Increased contact load and duration led to increased solid ECM strain and stress within, and increased ploughing force on the surface of the disc. CONCLUSION: Sustained mechanical loading may play a role in load carriage within the ECM and ploughing force formation during stress-field translation at the condyle-disc interface. This study further elucidated the mechanism of ploughing on tractional force formation and provided a baseline for future analysis of TMJ mechanics, cartilage fatigue and early TMJ degeneration.

A pilot study of nocturnal temporalis muscle activity in TMD diagnostic groups of women.

Temporomandibular disorder (TMD) incidences are believed to be related to parafunctional behaviours like teeth clenching. This pilot study aimed to (i) develop an automated clench-detection algorithm, and (ii) apply the algorithm to test for differences in nocturnal clenching in women with and without TMD. Subjects gave informed consent to participate. Adult women were categorised using Diagnostic Criteria for TMD according to presence/absence (+/-) of both TM joint disc placement (DD) and chronic pain (P) into two groups (+DD+P, -DD-P) with 12 subjects each. Surface temporalis electromyography was recorded during oral tasks performed by subjects at two laboratory sessions. The data were used to characterise muscle activity per N of bite force (µV/N) for each subject, develop the clench-detection algorithm and test its accuracy. Ambulatory surface temporalis electromyography was self-recorded by each subject over three nights and analysed using the algorithm and bite force (N) versus muscle activity µV/N calibrations. Bonferroni-adjusted homoscedastic t-tests assessed for significant between-group differences in clenching (P < 0·05). Sensitivity, specificity and accuracy of algorithm-detected laboratory clenches were all ≥96%. During self-recordings 95% of clenches had durations of <4 s and peak forces of <10 N in both groups. Mean clench durations were significantly longer (P = 0·042) in +DD+P (1·9 ± 0·8 s) than -DD-P subjects (1·4 ± 0·4 s). Mean temporalis duty factors (%clench time/total recording time) were significantly larger (P = 0·041) in +DD+P (0·47 ± 0·34%) than -DD-P (0·26 ±0·22%) subjects. Nocturnal temporalis muscle activities detected by a validated algorithm were longer per clench and recording time in +DD+P compared to -DD-P women.

Molecular characterization of a novel bipartite begomovirus isolated from Lycianthes biflora in China.

A bipartite begomovirus isolate GD was isolated from Lycianthes biflora plants showing yellow mosaic symptoms in Nanxiong, Guangdong Province, China. The apparently full-length DNA-A and DNA-B viral components were cloned after enrichment of circular DNA by rolling circle amplification, restriction digestion, cloning, and DNA sequencing. The DNA-A component (2752nt, KT582302) shares highest (80.2%) nucleotide (nt) sequence identity with tomato leaf curl Sulawesi virus [Indonesia-Sulawesi-Langowan F101-2006] (ToLCSuV- [ID-Sul -LanF09-06], FJ237618), reported in Indonesia as causing yellow leaf curl disease of chilli pepper. The DNA-B component (2704nt, KT582303) shares highest (76.3%) nt sequence identity with pepper yellow leaf curl Indonesia virus-[Indonesia-tomato2-2005] (PepYLCIV-[ID-Tom2-05 AB213599) reported in Indonesia, and associated with yellow leaf curl disease in tomato. Based on the ICTV guidelines for begomoviral species demarcation, the virus is a new, previously undescribed bipartite begomovirus species for which the name "Lycianthes yellow mosaic virus" is proposed.

[The corneal biomechanical properties of patients with Graves' orbitopathy].

OBJECTIVE: This study aims to compare the corneal biomechanical properties of patients with Graves'orbitopathy (GO) with normal people and analysis the correlated factors. METHODS: Retrospective case series study. 34 eyes of 20 patients (6 eyes are unilateral) diagnosed with inactive Graves' orbitopathy and 25 eyes of 14 patients (3 eyes are unilateral) diagnosed with active Graves' orbitopathy were recruited as study group. 30 eyes of 28 healthy volunteers with age and sex-matched were enrolled as control group in the affiliated eye hospital of Wenzhou medical university from December 2013 to June 2014. Central corneal thickness (CCT) measured with A ultrasound, corneal hysteresis (CH) measured with Ocular Response Analyser (ORA), corneal resistance factor (CRF), Goldman-corrected IOP value (IOPg) and Cornea-Compensated Intraocular Pressure(IOPcc) were collected. An ANONA and Kruskal-Wallis Test was used to compare the indexes mentioned above. RESULTS: The mean age of inactive group was (47.3±12.8)y (23 eyes were female, 11 eyes were male), the mean age of active group was (51.9±9.6)y (16 eyes were female, 9 eyes were male) , the mean age of normal group was (47.2±10.4)y (19 eyes were female, 9 eyes were male). CH of inactive group was (9.68±1.45) mmHg, (1 mmHg=0.133 kPa), CH of active group was (9.82±1.53) mmHg, lower than the CH of (10.67±1.68) mmHg in control group, which was statistically significant different (t1=-2.51,P1=0.014,t2=-2.01,P2=0.048). IOPcc of inactive group was (17.91±3.67) mmHg, IOPcc of active group was (17.88±5.44) mmHg, which were higher than the IOPcc of (13.90±3.39) mmHg in the control group, which was statistically significant different (t1=3.76,P1=0.001;t2= 3.461,P2=0.001). There were no significant difference between study and control group in CRF, that was in inactive group (10.19±1.73)mmHg, active group (10.36±1.01)mmHg, normal group (10.08±1.40)mmHg respectively (t1=0.31,P=0.761;t2=0.69,P2=0.491). CCT of inactive group was (531.41±37.60) µm, CCT of active group is (533.52±18.88) µm, which had no statistical significant difference from the CCT of (546.25±28.84) µm in control group (t1=-1.91,P=0.059;t2=-1.52,P2=0.132). Corneal hysteresis was negatively correlated with exophthalmos, IOPg, IOPcc (Pearson=-0.279,-0.385,-0.663,P<0.05), and positively correlated with corneal central thickness and corneal resistance factor (Pearson=0.246,0.583,P<0.05) in TAO group. CONCLUSIONS: Cornea hysteresis of TAO patients decreased, combined with lower ability of cornea to recover to the primary conditions when upon pressure. CH was negatively correlated with IOPg and IOPcc.(Chin J Ophthalmol, 2016, 52: 263-267).

[The effects of P - glycoprotein expression induced by ulinastatin on HK - 2 cells damage induced by paraquat].

Objective: To investigate the protective effect of P-glycoprotein up-regulated by ulinastatin (UTI) on HK-2 cells during paraquat (PQ) -induced injury and its underlying mechanisms. Methods: The re- search was divided into two parts. The first part of the research was divided into normal control group, PQ group, UTI+PQ group, UTI control group. The second part of the research was divided into negative virus group (including control group, PQ group, PQU+TI group, UTI group) and P-gp siRNA group (including control group, PQ group, PQU+TI group, UTI group) . Negative virus group: the cells were transfected into the blank virus; siRNA P-gp group: the cells were transfected with P-gp siRNA virus. HK-2 cells were routinely cultured. After 800 µmol/L PQ treatment, the changes of P-gp protein levels in the HK-2 cells were determined by West-ern-blot (WB) . Then, transfected lentivirus bringing P-gp silent gene, the cell viability was determined by CCK-8 assay, the expression of P-gp in the cells after transfection was detected by WB and the concentration of PQ in HK-2 cells were measured by high performance liquid chromatography (HPLC) . Results: Compared with the normal control group, the P-gp expression of PQ group had no significantly changes (P>0.05) . Compared with the PQ group, the P-gp expression of UTI+PQ group significantly increased (P>0.05) . Compared with the corre-sponding control siRNA group, the P-gp siRNA group had no significantly changes in cell viability (P>0.05) . and significantly decreased in P-gp expression. Compared with the corresponding control siRNA group, the P-gp siRNA group had no significantly changes in PQ concentration in HK-2 cell (P>0.05) , but compared with P-gp siRNA PQ group, the PQ concentration of P-gp siRNA PQ+UTI group significantly decrease (P<0.05) . Conclusion: UTI significantly reduced the accumulation of PQ in HK-2 cells and increased the viability of HK-2 cells in vitro may be not by increased P-gp activity. UTI could significantly reduce HK-2 cell injury induced by PQ in vitro and improve the survival rate of HK-2 cells. It may not be related to the up regulation of P-gp expres-sion.

Salmonella L-forms: formation in human bile in vitro and isolation culture from patients' gallbladder samples by a non-high osmotic isolation technique.

Bacterial L-forms have always been considered as osmotic-pressure-sensitive cell-wall-deficient bacteria and isolation culture of L-forms must use media with high osmotic pressure. However, isolation culture of stable L-forms formed in humans and animals is very difficult because they have adapted to the physiological osmotic pressure condition of the host. We use a non-high osmotic isolation technique to isolate stable L-forms of Salmonella Typhi and Salmonella Paratyphi A from bile-inducer cultures in vitro and from patients' gallbladder specimens. Multiplex PCR assay for Salmonella-specific genes and nucleotide sequencing are used to identify the Salmonella L-forms in stable L-form isolates. Using this method, we confirmed that Salmonella Paratyphi A and Salmonella Typhi cannot be isolated from bile-inducer cultures cultured for 6 h or 48 h, but the L-forms can be isolated from 1 h to 45 days. In the 524 gallbladder samples, the positive rate for bacterial forms was 19.7% and the positive rate for Salmonella spp. was 0.6% by routine bacteriological methods. The positive rate for bacterial L-forms was 75.4% using non-high osmotic isolation culture. In the L-form isolates, the positive rate of Salmonella invA gene was 3.1%. In these invA-positive L-form isolates, four were positive for the invA and flic-d genes of Salmonella Typhi, and ten were positive for the invA and flic-a genes of Salmonella Paratyphi A.

Upregulation of cytoskeleton protein and extracellular matrix protein induced by stromal-derived nitric oxide promotes lung cancer invasion and metastasis.

Lung cancer commonly metastasizes to lymph nodes, brain and bones, which is the main cause of death. It is still a challenge to detect molecular biomarkers for early diagnosis and therapeutics of lung cancer. Our previous study found that bone marrow-derived stroma cells (BMSCs) under tumor microenvironment produced nitric oxide (NO), which was induced by inducible nitric oxide synthase (iNOS), and promoted invasion and metastasis of cancer cells by remodeling cytoskeleton. The aim of this study is to elucidate the relationship between the expressions of iNOS, cytoskeleton protein caldesmon, OPN, and clinical parameters especially the metastasis of lung cancer. We found that nitric oxide can remodel cytoskeleton and promoted the mobility of lung cancer cells. The expressions of iNOS, caldesmon, and OPN are closely correlated to metastasis of lung cancer. The intracranial metastatic tissue samples of lung cancer showed significantly higher expression of iNOS, caldesmon and OPN. A flow-cytometry analysis for peripheral blood of lung cancer patients showed increased EPCAM+/OPN+ cells in circulation of patients with bone metastasis compared to that of patients without metastasis, which is indicative of cancer circulating cells. The concentration of serum OPN was also positively related to the bone metastasis of lung cancer. Taken together, these results suggested that iNOS, caldesmon and OPN may work as biomarkers for metastasis of lung cancer.

Identification and molecular characterization of two begomoviruses from Pouzolzia zeylanica (L.) Benn. exhibiting yellow mosaic symptoms in adjacent regions of China and Vietnam.

Two monopartite begomoviruses were isolated from Pouzolzia zeylanica (L.) Benn. plants showing yellow mosaic symptoms in Gaoyao, Guangdong Province, China (GD1) and in Phu Tho, Vietnam (VN), respectively. A comparison of the complete genome sequence of GD1 (2,739 nucleotides [nt]) with VN (2,741 nt) indicated that they shared 86.2 % nt sequence identity. GD1 and VN shared the highest nucleotide sequence identity at 86.7 % and 91.4 % respectively, with isolate TY01 of pouzolzia golden mosaic virus (PGMV-TY01), another begomovirus isolated from P. zeylanica. Phylogenetic analysis revealed that GD1, VN, and PGMV-TY01 were members of a distinct begomovirus clade. Based on the ICTV guidelines for begomoviral species demarcation, GD1 belongs to a new begomovirus species, for which the name Pouzolzia yellow mosaic virus is proposed. Likewise, VN represents a previously unreported strain of PGMV. Recombination analysis predicted that VN was a recombinant between PGMV-TY01 and ageratum yellow vein China virus isolate G13 (AYVCNV-G13), and that PGMV-TY01 and VN were likely the parents of GD1 through recombination with allamanda leaf curl virus isolate G10 (AlLCV-G10), a begomovirus endemic to Guangdong Province of China.

First Report of Sweet potato leaf curl Georgia virus Infecting Tall Morning Glory (Ipomoea purpurea) in China.

In September 2013, tall morning glory (Ipomoea purpurea) plants showing vein yellowing and leaf curl symptoms typical of a begomovirus infection were observed in Jingzhou, Hubei Province, China. Total nucleic acids were extracted from a symptomatic plant using cetyltrimethylammonium bromide (CTAB). Rolling circle amplification (RCA) was conducted using TempliPhi kit (GE Healthcare) to recover the genome of a putative begomovirus. Digestion of the RCA product with PstI yielded a ~2.8 kbp DNA fragment suggestive of a monomerized begomoviral genome. The fragment was cloned and sequenced and the sequence was deposited in GenBank under accession no. KF769447. SDTv1.0 (species demarcation tool) analysis revealed that the putative begomovirus showed 98.5 and 92.0% nucleotide sequence identity with Sweet potato leaf curl Georgia virus (SPLCGV)-[China:Hebei:2011] (GenBank Accession No. JX448368) and SPLCGV-[US:Geo:16] (AF326775), respectively. The virus contained six ORFs, which encoded proteins showing 96.5 to 100% and 90.6 to 95.6% amino acid sequence identity with their counterparts of SPLCGV-[China:Hebei:2011] and SPLCGV-[US:Geo:16], respectively. Thus, the virus should be considered as an isolate of SPLCGV-[China:Hebei:2011]. Tall glory morning in a nearby field (which covers an area of 3 square kilometers) was surveyed and 70 to 100% of plants were found showing symptoms reminiscent of begomoviral infection. Total nucleic acid was extracted from 13 randomly selected (10 symptomatic and 3 healthy) plants and used as templates for PCR with a pair of specific primers (5'-CGCAGCCTTTCCACACTATC-3'/5'-AAAACAGTTTGGGCTCGGTC-3') designed according to the sequence described above. Positive results were obtained for all of the symptomatic, but none of the healthy-looking tall morning glory plants. SPLCGV (genus Begomovirus, family Geminiviridae) was reported to infect sweet potato (I. batatas) in the United States (4), India (2), and China (3). To our knowledge, this is the first report of SPLCGV infecting tall morning glory in China. Also, it is the first report of a geminivirus in Hubei, a province of central China. Whereas the finding of SPLCGV in sweet potato (3) may be a result of vegetative propagation of this crop, the detection of SPLCGV in tall morning glory, an annual plant, raises the possibility that this virus is transmissible and is spreading in China. References: (1) B. Muhire et al. Arch. Virol. 158:1411, 2013. (2) G. Prasanth and V. Hegde. Plant Dis. 92:311, 2008. (3) Y. Qin et al. Plant Dis. 97:1388, 2013. (4) R. A. Valverde and D. L. Gutierrez. Rev. Mex. Fitopatol. 21:128, 2003.

First Report of Wild tomato mosaic virus Infecting Tobacco (Nicotiana tabacum) in China.

Wild tomato mosaic virus (WTMV), a potyvirus, has been reported in Laichau, Vietnam, infecting Solanum torvum (wild tomato) in 2008 (3), and Kanchanaburi, Thailand, infecting Capsicum spp. in 2013 (KF250353). In mid-May 2013, Nicotiana tabacum showing yellowing, mosaic, and/or ringspot symptoms were found in natural tobacco fields of Nanxiong, Guangdong Province, China. Total RNA was extracted from symptomatic leaves and reverse transcribed with M4T (5'-GTTTTCCCAGTCACGAC (T)15-3') as the 3' anchoring primer (1). The cDNA was used as template in a PCR assay using primers M4: 5'-GTTTTCCCAGTCACGAC-3' and Sprimer: 5'-GGXAAYAAYAGYGGXCAZCC-3', which amplifies a region comprising part of the NIb protein gene, the entire coat protein (CP) gene and the 3' nontranslated region (UTR) of a potyvirus (1). A ~1,700-bp product was amplified from the cDNA derived from three of the five diseased plants. The product (KF639967) showed 87% and 84% nucleotide sequence identities with those of WTMV isolates KAN and Laichau, respectively. The CP deduced from the sequence of the product shared 87% and 86% nucleotide and 94% and 93% amino acid sequence identities with those of WTMV isolates KAN and Laichau, respectively. The 3'-UTR of the putative virus shared 93% and 92% nucleotide sequence identities to those of WTMV isolates KAN and Laichau, respectively. Thus, according to the molecular criteria for potyvirus species demarcation (2), the virus we identified should be an isolate of WTMV (isolate GD1). One of the diseased samples was homogenized in 0.1 mol/liter phosphate buffer (pH 7.0) and used to inoculate the potyvirus to healthy, two to four leaf-stage Capsicum annuum L., N. tabacum, and N. benthamiana. The inoculated, as well as mock-treated plants, which were inoculated only with phosphate buffer, were grown in soil under 12 h day/12 h night at 25°C. All inoculated N. tabacum and N. benthamiana plants developed yellowing and mosaic symptoms by 14 days post inoculation (dpi). For N. benthamiana, the symptom became very severe by 21 dpi and some diseased plants died prematurely. About 10% of inoculated C. annuum L. developed very mild veinal chlorosis 18 dpi. Cloning and sequencing experiments showed that all the symptomatic plants tested were WTMV positive, but Cucumber mosaic virus, Tobacco mosaic virus, and Tobacco etch virus negative. To our knowledge, this is the first report of WTMV in China. Also, it is the first report that WTMV infects Nicotiana spp. Although further experiments are needed to definitively attribute the disease observed in the field to WTMV, our results indicate that WTMV, which forms a monophyletic clade with a number of other potyviruses infecting Solanaceae species in phylogenetic analysis, is widely distributed, or is spreading in Southeast Asia. It may pose a threat to Solanaceae species cultivation in this region. References: (1) Chen et al. Arch. Virol. 146:757, 2001. (2) Adams et al. Arch. Virol. 150:459, 2005. (3) Ha et al. Arch. Virol. 153:25, 2008.

Late, severe, noninfectious diarrhea after renal transplantation: high-risk factors, therapy, and prognosis.

OBJECTIVE: Late severe noninfectious diarrhea in renal transplant recipients can lead to malnutrition and even graft loss. The purpose of this study was to evaluate risk factors associated with this condition and summarize therapy for these patients. METHODS: For more than 36 months we observed a cohort of 541 recipients who underwent kidney transplantation from January 2001 to June 2007. They were provided a calcineurin inhibitor (CNI) combined with mycophenolate mofetil (MMF). The four group includes a continuous cyclosporine (CsA); a preconversion to tacrolimus and a postconversion group as well as a continuous tacrolimus group. The rate of severe late noninfectious diarrhea was compared among the four groups. Risk factors were analyzed between the diarrhea and nondiarrhea cohorts. Clinical characteristics, efficacy, and safety were observed after modifying the immunosuppressive protocol for late severe noninfectious diarrhea recipients. RESULTS: Twenty-eight recipients presented with late sever noninfectious diarrhea. No patients displayed chronic diarrhea in the CsA (n = 145) or preconversion group (n = 95). The rate of diarrhea was 7.31% in the postconversion and 7.35% in the tacrolimus group. Using multivariate Cox proportional hazards analysis, factors associated with an increased risk of noninfectious diarrhea were cytochrome P450(CYP)3A5 *3/*3 type, chronic renal allograft dysfunction, and patient ingestion of Tripterygium wilfordii Hook F. All diarrheal recipients experienced weight loss, hypoalbuminia, and an increased serum creatinine. All affected patients underwent adjustment of the immunosuppressive regimen to achieve remission. Renal allograft survival in recipients with diarrhea was worse than that in nondiarrheal recipients receiving tacrolimus combined with MMF. CONCLUSION: Tacrolimus with MMF increased the risk of late severe noninfectious diarrhea among renal transplant recipients compared with hosts treats with CsA plus MMF. The CYP3A5 *3/*3 type, chronic renal allograft dysfunction, and T. wilfordii supplementation were high-risk factors for late diarrhea. Prompt adjustment of immunosuppression was an effective, feasible therapy for these patients.

Molecular characterization of a novel monopartite begomovirus isolated from Pouzolzia zeylanica in China.

The complete genome sequence of a monopartite begomovirus isolate TY01 was obtained from diseased Pouzolzia zeylanica plants exhibiting golden mosaic symptoms in Baise, Guangxi Province, China. It consisted of 2723 nucleotides (nt) and encoded two ORFs (CP and AV2) in the virion-sense DNA and five ORFs (AC1-AC5) in the complementary-sense DNA. Compared with the DNA-A sequences of other begomoviruses, it has the highest (78.5 %) nucleotide sequence identity with ageratum yellow vein virus (AYVV) isolate AFSP6D from Thailand, which is less than the 89 % identity in the complete genome that has been defined as the threshold value for demarcation of species in the genus Begomovirus, family Geminiviridae. Phylogenetic analysis showed that TY01 was grouped in a separate clade from the other 28 begomovirus isolates. These results indicate that isolate TY01 is a member of a novel Begomovirus species, for which the name "Pouzolzia golden mosaic virus" (PGMV) is proposed.

First Report of Potato Blackleg Disease Caused by Pectobacterium atrosepticum in Guangdong China.

Potato (Solanum tuberosum L.) is an important crop in China. In 2013, diseased potatoes exhibiting blackleg and soft rot symptoms were found in the winter potato growing areas of Huizhou city, Guangdong Province, China, with an incidence of approximately 20%. Initially, the stem bases of infected plants blackened and this symptom spread upward. Later, foliage of the diseased plants became yellow and the stem rotted with vascular discoloration. Twenty diseased plants with typical black leg symptoms were collected from a 10-ha potato field with approximately 60,000 potato plants per hectare. A bacterium with small, irregular, round, fluidal, white colonies was isolated from the vascular tissue of all diseased plants on nutrient agar at 26°C for 2 days. Ten strains were randomly selected for pathogenicity assays. Potato plants (cv. Favorita) at the five- to six-leaf stage were inoculated by injecting their stems with 1 ml of each strain in a bacterial suspension (3 × 108 CFU/ml). The inoculated potato plants were incubated at 16 to 21°C and 65 to 85% humidity, and exhibited the same symptoms as the diseased potato plants in the field by 3 to 5 days post inoculation (dpi). The bacterium was reisolated from the diseased tissue (stem) of the inoculated potato plants and produced characteristic pits on crystal violet pectate medium (1). The bacterium utilized a-methyl glucoside, glucose, lactose, maltose, cellobiose, raffinose, melibiose, and citrate, but not d-arabitol, sorbitol, or malonate. The bacteria also gave a positive reaction for catalase and production of reducing substances from sucrose, but gave a negative reaction for oxidase, production of phosphatase, and indole. Using the universal bacterial 16S rDNA primer set, 27f/1541R (4), 1,400-bp fragments were amplified from the 10 strains. The sequences of the 10 fragments (GenBank Accessions KC695819 to KC695828) were identical and had 100% sequence identity with 16S rDNA of Pectobacterium atrosepticum CFBP 1526 (JN600332). Further, the 438-bp and 690-bp fragments were respectively amplified from all 10 strains with the P. atrosepticum-specific primers Y45/Y46 (3) and ECA1f/ECA2r (2). To our knowledge, this is the first report of potato blackleg disease caused by P. atrosepticum (formerly named as Erwinia carotovora subsp. atroseptica) in Guangdong Province, China. References: (1) D. Cupples et al. Phytopathology 64:468, 1974. (2) S. H. De Boer et al. Phytopathology 85:854, 1995. (3) D. Frenchon et al. Potato Research 41:63, 1995. (4) M. Horita et al. J. Gen. Plant Pathol. 70:278, 2004.

First Report of Bacterial Wilt Caused by Ralstonia solanacearum on Ageratum conyzoides in China.

Ageratum conyzoides L. is believed to act as reservoir host for many plant diseases. In June 2011, a 30% incidence of bacterial wilt on A. conyzoides was observed in a field of Rhizoma kaempferiae in Yangchun city of Guangdong province. The initial symptoms were wilting of the apical leaves during the day, which recovered at night. After 4 to 6 days, the leaves became totally necrotic. The basal stems of the diseased plants were blackened and the vascular tissue turned brown. To investigate the disease etiology before understanding the disease link between A. conyzoides and R. kaempferiae, 10 plants with typical wilting symptoms were collected from the field. A total of 10 bacterial isolates were isolated from the vascular tissue of each diseased plant on tripheny tetrazolium chloride (TZC) medium. After incubation at 30°C for 2 days, the plates had large, irregular round, fluidal, white colonies with a pink center. Thirty healthy A. conyzoides plants at the four- to six-leaf growth stage were inoculated by injuring the roots and soaking them in a bacterial suspension (1 × 108 cfu/ml) for 20 min with the 10 bacterial isolates separately, and planted in 10-cm pots with sterile gardening soil in a glasshouse (28 to 35°C). Sterile water was used as a negative control. Five days after inoculation, a few leaves of the inoculated plants began to exhibit wilting. The inoculated plants eventually showed the same symptoms as those in the field. The same bacterium was reisolated from inoculated plants. The 30 negative control plants did not have wilt symptoms. With the same inoculation procedure, the bacterium also caused wilting on tomato (25 of 30), pepper (10 of 30), eggplant (2 of 30), ginger (11 of 15), and R. kaempferiae (8 of 15). Using the universal bacterial 16S rDNA primer set 27f/1541R (3), approximately 1,400 bp-fragments were amplified from the 10 isolates, respectively. The sequences for the 10 fragments (GenBank Accession Nos. JX294065 to JX294074) were identical and had 100% sequence identity with 16S rDNA of R. solanacearum GMI1000 (AL646052). The 10 isolates were able to oxidize disaccharides (lactose, maltose, and cellobiose) and hexose alcohols (mannitol, dulcitol, and sorbitol). According to Hayward's classification, all isolates were biovar 3 (2). Based on the pathogenicity tests, carbohydrate utilization, and near full-length 16S rDNA sequences, the bacterial isolates from the diseased A. conyzoides belonged to race 4 and biovar 3 of R. solanacearum. Furthermore, the specific 280-bp and 140-bp fragments were respectively amplified from all 10 isolates by using the multiplex PCR (1). In addition, specific 165-bp fragments were amplified from all the isolates using the specific primers AKIF/AKIR (3), which indicates the bacterium belongs to R. solanacearum Phylotype I. To our knowledge, this is the first report of a disease caused by R. solanacearum on A. conyzoides in China. References: (1) M. Fegan and P. Prior. Page 449 in: Bacterial Wilt Disease and the Ralstonia solanacearum Species Complex. C. Allen et al., eds. The American Phytopathological Society. St. Paul, MN, 2005. (2) A. C. Hayward. J. Appl. Bacteriol. 27:265, 1964. (3) M. Horita et al. J. Gen. Plant Pathol. 70:278, 2004.