Secondary metabolites isolated from Trichoderma hamatum b-3 and their fungicidal activity.

An undescribed trichodenone derivative (1), two new diketopiperazines (3 and 4) along with a bisabolane analog (2) were isolated from Trichoderma hamatum b-3. The structures of the new findings were established through comprehensive analyses of spectral evidences in HRESIMS, 1D and 2D NMR, Marfey's analysis as well as comparisons of ECD. The absolute configuration of 2 was unambiguously confirmed by NMR, ECD calculation and Mo2(AcO)4 induced circular dichroism. Compounds 1-4 were tested for their fungicidal effects against eight crop pathogenic fungi, among which 1 showed 51% inhibition against Sclerotinia sclerotiorum at a concentration of 50 µg/mL.

Time course analysis of sensory axon regeneration in vivo by directly tracing regenerating axons.

Most current studies quantify axon regeneration by immunostaining regeneration-associated proteins, representing indirect measurement of axon lengths from both sensory neurons in the dorsal root ganglia and motor neurons in the spinal cord. Our recently developed method of in vivo electroporation of plasmid DNA encoding for enhanced green fluorescent protein into adult sensory neurons in the dorsal root ganglia provides a way to directly and specifically measure regenerating sensory axon lengths in whole-mount nerves. A mouse model of sciatic nerve compression was established by squeezing the sciatic nerve with tweezers. Plasmid DNA carrying enhanced green fluorescent protein was transfected by ipsilateral dorsal root ganglion electroporation 2 or 3 days before injury. Fluorescence distribution of dorsal root or sciatic nerve was observed by confocal microscopy. At 12 and 18 hours, and 1, 2, 3, 4, 5, and 6 days of injury, lengths of regenerated axons after sciatic nerve compression were measured using green fluorescence images. Apoptosis-related protein caspase-3 expression in dorsal root ganglia was determined by western blot assay. We found that in vivo electroporation did not affect caspase-3 expression in dorsal root ganglia. Dorsal root ganglia and sciatic nerves were successfully removed and subjected to a rapid tissue clearing technique. Neuronal soma in dorsal root ganglia expressing enhanced green fluorescent protein or fluorescent dye-labeled microRNAs were imaged after tissue clearing. The results facilitate direct time course analysis of peripheral nerve axon regeneration. This study was approved by the Institutional Animal Care and Use Committee of Guilin Medical University, China (approval No. GLMC201503010) on March 7, 2014.

Polycomb protein family member CBX7 regulates intrinsic axon growth and regeneration.

Neurons in the central nervous system (CNS) lose their intrinsic ability and fail to regenerate, but the underlying mechanisms are largely unknown. Polycomb group (PcG) proteins, which include PRC1 and PRC2 complexes function as gene repressors and are involved in many biological processes. Here we report that PRC1 components (polycomb chromobox (CBX) 2, 7, and 8) are novel regulators of axon growth and regeneration. Especially, knockdown of CBX7 in either embryonic cortical neurons or adult dorsal root ganglion (DRG) neurons enhances their axon growth ability. Two important transcription factors GATA4 and SOX11 are functional downstream targets of CBX7 in controlling axon regeneration. Moreover, knockdown of GATA4 or SOX11 in cultured DRG neurons inhibits axon regeneration response from CBX7 downregulation in DRG neurons. These findings suggest that targeting CBX signaling pathway may be a novel approach for promoting the intrinsic regenerative capacity of damaged CNS neurons.

Lethal/sublethal responses of Daphnia magna to acute norfloxacin contamination and changes in phytoplankton-zooplankton interactions induced by this antibiotic.

Although the well-known antibiotic norfloxacin (NOR) is recognized as an important environmental pollutant, little is known about its impacts on ecological processes, particularly on species interactions. In this paper, we quantified Daphnia magna (Crustacea, Cladocera) responses in mortality rate at lethal NOR concentrations (0, 25, 50, 100, 200, 300 and 400 mg L-1), and in heartbeat rate, swimming behavior and feeding rate (on the green alga Chlorella pyrenoidosa) at sublethal NOR concentrations (0, 25, 50 and 100 mg L-1) to determine the effects of this antibiotic in plankton systems. In 96-h-long lethal experiment, mortality rates of D. magna increased significantly with increasing NOR concentration and exposure time. In sublethal experiments, heartbeat rate decreased, while time ratio of vertical to horizontal swimming (TVH) and the duration of quiescence increased in D. magna individuals exposed to increasing NOR concentrations after 4 and 12 h of exposure. These collectively led to decreases in both average swimming ability and feeding rate, consistent with the positive relationship between average swimming ability and feeding rate. Overall, results indicate that, by affecting zooplankton heartbeat rate and behavior, NOR decreased feeding efficiency of D. magna even at low doses, therefore, it might seriously compromise ecosystem health and function.

MicroRNA-210 promotes sensory axon regeneration of adult mice in vivo and in vitro.

Axon regeneration as a critical step in nerve repairing and remodeling after peripheral nerve injury relies on regulation of gene expression. MicroRNAs are emerging to be important epigenetic regulators of gene expression to control axon regeneration. Here we used a novel in vivo electroporation approach to transfect microRNA-210 (miR-210) or siRNAs to adult mice dorsal root ganglion (DRG) neurons, measured the axon length 3days after sciatic nerve crush or dissociated DRG cultures in vitro to detect the effect of miR-210 in sensory axon regeneration. Importantly, we found that miR-210 overexpression could promote sensory axon regeneration and inhibit apoptsosis by ephrin-A3 (EFNA3). In addition, inhibition of endogenous miR-210 in DRG neurons impaired axon regeneration in vitro and in vivo, the regulatory effect of miR-210 was mediated by increased expression of EFNA3 because downregulation of EFNA3 fully rescued axon regeneration. We thus demonstrate that miR-210 is a new physiological regulator of sensory axon regeneration, and EFNA3 may be the functional target of miR-210. We conclude that miR-210 may play an important role in sensory axon regeneration.

Study on optimization of proportion between fermented liquid and traditional cultural medium of bioflocculant production and its flocculant performance considering the aerobic fermentation of rice straw as substrate.

High cost of traditional culture medium of flocculant is the key element to limit the bioflocculant production. It's therefore much crucial to seek the economic production materials. In this research, part of the traditional culture medium of bioflocculant is replaced by the fermented liquid of rice straw to conduct the discussion on fermentation matching, optimization of fermentation condition and ability of flocculant production. The optimal proportion of aerobic saccharification liquid and traditional cultural medium of flocculant production is 1: 3. The flocculant rates of the economic culture medium of flocculant production are the highest, 65.49% and 71.24%, which are combined by 67d and 109d fermented saccharification liquid and the traditional cultural medium of flocculant production. The growth of flocculant production bacterium is in better situation for composite culture medium of flocculant production. The amount of bioflocculant is 40kg from per ton. The fermentation cost of flocculant saves by 25% comparing with the traditional culture medium. The simple aerobic fermentation technique opens up a new road for low-cost culture medium of flocculant production.

Temperature-insensitive detection of low-concentration nanoparticles using a functionalized high-Q microcavity.

The ability to detect nanoparticles in extremely dilute solutions in the presence of environmental noise is crucial for biosensing applications. In this paper we propose a scheme for detecting target nanoparticles through their scattering effects in a high-Q whispering gallery microcavity. The detection signal, defined as the total linewidth broadening of the two new split modes that appear upon nanoparticle adsorption, is highly sensitive and proportional to the nanoparticle concentration. Furthermore, this new method of detection eliminates the requirement for strict temperature control and is capable of distinguishing the signal from the biorecognitions (e.g., antibodies) initially attached to the resonator and that from the target nanoparticles (e.g., antigens).

[Evaluation the value of Pentacam arterial segment analysis system in the study of anterior segment of the eye before and after laser periphery iridectomy].

OBJECTIVE: To evaluate the anterior segment parameters in early primary angle-closure glaucoma (PACG) and primary angle closure (PAC) eyes before and after neodymium:yttrium aluminum garnet (Nd:YAG) laser periphery iridectomy (LPI) by using Pentacam system. METHODS: Self-control study. Sixty one eyes of 33 patients were recruited into this study. The images of the anterior segment of the eye were captured and the parameters [i.e. the anterior chamber (AC) volume, the central AC depth, the peripheral AC depth, the peripheral AC angle and the pupil size] were measured by the Pentacam system pre-operatively and 1 week after laser treatment. RESULTS: The Pentacam system clearly showed the structure changes of the anterior segment after LPI (i.e. the convex configuration of the iris was flattened and the peripheral AC depth was deepened). The average AC volume (t=15.928), the peripheral AC depth (t=-17.399, -10.629, -12.576, -14.474) and the peripheral AC angle (t=-5.087) were increased significantly (P<0.01) after LPI. The AC volume was (74.10±16.35) mm3 before LPI and (94.10±18.42) mm3 after LPI, and the increases of peripheral AC depth were 0.17 mm (nasal) and 0.29 mm (temporal). There was no statistical significant changes in the AC depth before and after LPI. CONCLUSION: In patients with PACG and PAC, the changes of AC after LPI can be demonstrated by the Pentacam qualitatively and quantitatively.

[Measurement of central corneal thickness in myopic eyes with ultrasound and Pentacam scheimpflug system].

OBJECTIVE: To assess the accuracy and variability of central corneal thickness measurements in myopic eyes obtained by ultrasound pachymetry and the Pentacam scheimpflug imaging system. METHODS: First, corneal thickness in 50 healthy eyes was measured three times with the Pentacam system to evaluate the repeatability. Then, Scheimpflug imaging system (Pentacam) and A-scan ultrasonic pachymeter (SP-3000) were used to measure the corneal thickness on 217 myopic cases (433 eyes) undergoing laser in situ keratomileusis (LASIK) and 63 cases (126 eyes) 6 months after LASIK. The results were compared. RESULTS: Repeated-measures ANOVA showed the repeatability of the Pentacam measurements was high (P > 0.05). Pre-operatively, the value of corneal central thickness measured by ultrasonic pachymetry and the Pentacam system was (537.26 +/- 31.61) microm and (538.08 +/- 32.72) microm, respectively. There was no significant difference between the results of these two methods (t = 1.867, P > 0.05). The difference between these two methods ranged -35 - 34 microm, and was within -20 - 20 microm in 95.38% cases. Post-operatively, the value of corneal central thickness measured by ultrasonic pachymetry and the Pentacam system was (464.21 +/- 41.04) microm and (465.17 +/- 42.65) microm, respectively. There was no significant difference between these two methods, too (t = 0.913, P > 0.05). There was a significant linear correlation between the two methods (P < 0.001). The thinnest point of cornea was mostly located surrounding the apex and inclined towards inferotemporal. CONCLUSION: The Pentacam scheimpflug system offers an acceptable method for evaluating corneal pachymetry, which is useful in corneal refractive surgery.

[Hemostatic effects of Sorbalgon dressing after intranasal endoscopic surgery].

OBJECTIVE: To study the therapeutic effects of Sorbalgon dressing for hemostasis after intranasal endoscopic surgery. METHODS: Intranasal endoscopic surgery was performed in 50 patients with bilateral chronic nasosinusitis and nasal polyps, after which the hemostatic effect of Sorbalgon and Vaseline dressings and nasal cavity reaction were observed. RESULTS: Sorbalgon dressing resulted in milder nasal swelling and pain without obvious nasal hemorrhage and mucous membrane reaction, and the average time of nasal recovery was shorter in comparison with Vaseline dressing, which caused severe pain, obvious haemorrhage, and nasal mucous membrane swelling with prolonged nasal recovery. Three months after the operation, similar mucous membrane epithelialization was observed in the nasal cavity managed with the two dressings. CONCLUSION: Sorbalgon dressing has good hemostatic effect after intranasal endoscopic surgery.

[Clinical features of 23 cases of AIDS complicated by tuberculosis].

OBJECTIVE: To discuss the clinical features, treatment and prognosis of patients with AIDS complicated by tuberculosis(TB). METHODS: The clinical features of 23 patients with AIDS complicated by tuberculosis admitted from 1997 to July 2004 were retrospectively analyzed. RESULTS: Of the 23 patients, most (94.3%) were young or middle-aged, and 11 (47.8%) died within half a year. The main HIV transmission was via sexual contact in 15 (65.2%) patients. Loss of body weight by 5 - 15 kg was present in all patients, cough for over 1 month in 15 (65.2%), and multiple opportunistic infections were complicated in most cases. Out of the 23 cases, 14 (60.9%) showed only pulmonary TB, and 8 (34.8%) showed lymph node TB. In 12 cases with infiltrated pulmonary TB, X-ray showed bilateral infiltration and no cavity formation was found. Slightly positive PPD test was found in 2 (8.7%) cases, and positive acid-fast bacilli was detected in sputum in 1 case (4.4%). The pre-treatment CD(4)(+) cell number in 23 patients was much lower than that in AIDS patients without complicated TB (P < 0.05). The pre-treatment CD(4)(+) cell number in patients died shortly after diagnosis was much lower than that in survived patients (P < 0.05). The HIV RNA level in the 23 patients was much higher than that in patients without complication of TB (P < 0.05). The mortality in patients treated with therapy against both TB and HIV was much lower than that in patients untreated or treated only with anti-TB therapy (P < 0.05). CONCLUSIONS: For patients with AIDS complicated by TB, the high negative rate in PPD test and the atypical chest X-ray manifestations are common. However, lymph node TB is quite common with high mortality. The pre-treatment CD(4)(+) level decreases significantly, and is associated with mortality. The TB bacilli may accelerate HIV virus duplication. It is suggested that the patients be treated with a combination of anti-TB and anti-HIV therapies.

[Identification and molecular cloning and sequence analysis of a novel coronavirus from patients with SARS by RT-PCR].

OBJECTIVE: To investigate the etiologic agents of the SARS and develop diagnostic method for this disease. METHODS: Thirty-six nasopharyngeal aspirate specimens from 27 patients with SARS in Shenzhen were collected. The samples were aliquotted to three parts and subjected to molecular assays for human metapneumovirus, chlamydia and a novel coronavirus, which was reported recently to be the etiologic agent of SARS. Nested RT-PCR was used to amplify the RNA polymerase gene of the novel coronavirus and the PCR products were sequenced directly or after cloned to pMD18-T vector. RESULTS: Human metapneumovirus and chlamydia genes were detected in none of the specimens using the RT-PCR and nested-PCR, respectively. The novel coronavirus gene were amplified in 6 of 36 specimens, the sequence analysis indicated that this novel coronavirus is unrelated to any other coronavirus reported previously. The nucleotide and deduced amino acid alignment between this coronavirus and others was not more than 40% and 70% to 82%, respectively, while the nucleotide sequence cloned from the 6 patients were identical. CONCLUSIONS: The SARS patients in Shenzhen were infected with coronavirus and this novel coronavirus is associated with SARS. The sequence analysis indicated that the coronavirus from SARS patients in Shenzhen is the same as that identified from other areas such as Canada and Hong Kong. A specific diagnostic nested RT-PCR was developed to identify this novel coronavirus infection.

[Clinical and chest X-ray characteristics of 5 cases with severe acute respiratory syndrome in children in Shenzhen area].

OBJECTIVE: To explore clinical and chest X-ray features of SARS in children to facilitate correct diagnosis. METHODS: Clinical manifestations and chest X-ray findings in five children suffering from SARS admitted for treatment in the hospital between February and May, 2003 in Shenzhen area were analyzed. The diagnosis was confirmed by epidemiological, clinical, laboratory and radiological examinations. Among the 5 cases, 1 was a boy and the others were girls at the age of 4 to 13 years. RESULTS: Of the 5 SARS children, 3 presented a history of close contact with SARS patients. Fever was the initiative symptom, 4 had a body temperature of over 38 degrees C with the highest being 40 degrees C; fever sustained from 4 to 7 days with an average of 5.6 days. All the 5 cases developed nonproductive cough; on auscultation, both moist and dry rales could be heard in 3 out of the 5 cases. Mean total white count of peripheral blood was (2.96 - 6.9) x 10(9)/L, and was < 5.0 x 10(9)/L in 4 cases. SARS associated coronavirus specific RNA fragment was found positive by RT-PCR in 1 case; 1 case was positive for both IgM and IgG antibodies to the virus; 1 case was positive for only IgM antibody and another 2 cases were positive for only IgG antibody. IgG and IgM antibodies to Mycoplasma pneumoniae and Chlamydia pneumoniae as well as blood culture for bacteria were all negative. Findings on chest X-ray examination: 4 cases showed presence of patchy or macular opacities with cord-like shadows in unilateral lung plates while 1 case each showed ground-glass-like opacity and migratory changes; 1 case showed interstitial changes in the lungs in the form of irregular reticular lattice and cord-like shadows. Two cases received CT scanning and macular-patchy or spotty shadows were seen all over the lung. The shortest time for absorption of foci in the lungs was 7 days while the longest was 33 days with a mean of 15 +/- 6 days. None of the cases had any signs of fibrosis in the lungs. All the 5 cases were completely cured and discharged 7 to 40 days (mean 18 +/- 11 days) after admission. CONCLUSION: Compared with adult cases with SARS, children with SARS had milder symptoms and signs. Presence of unilateral patchy shadow in lungs represented the main chest X-ray findings.