Ocular A-to-I RNA editing signatures associated with SARS-CoV-2 infection.

Ophthalmic manifestations have recently been observed in acute and post-acute complications of COVID-19 caused by SARS-CoV-2 infection. Our precious study has shown that host RNA editing is linked to RNA viral infection, yet ocular adenosine to inosine (A-to-I) RNA editing during SARS-CoV-2 infection remains uninvestigated in COVID-19. Herein we used an epitranscriptomic pipeline to analyze 37 samples and investigate A-to-I editing associated with SARS-CoV-2 infection, in five ocular tissue types including the conjunctiva, limbus, cornea, sclera, and retinal organoids. Our results revealed dramatically altered A-to-I RNA editing across the five ocular tissues. Notably, the transcriptome-wide average level of RNA editing was increased in the cornea but generally decreased in the other four ocular tissues. Functional enrichment analysis showed that differential RNA editing (DRE) was mainly in genes related to ubiquitin-dependent protein catabolic process, transcriptional regulation, and RNA splicing. In addition to tissue-specific RNA editing found in each tissue, common RNA editing was observed across different tissues, especially in the innate antiviral immune gene MAVS and the E3 ubiquitin-protein ligase MDM2. Analysis in retinal organoids further revealed highly dynamic RNA editing alterations over time during SARS-CoV-2 infection. Our study thus suggested the potential role played by RNA editing in ophthalmic manifestations of COVID-19, and highlighted its potential transcriptome impact, especially on innate immunity.

Case Report: Molecular and microenvironment change upon midostaurin treatment in mast cell leukemia at single-cell level.

Mast cell leukemia is a rare and aggressive disease, predominantly with KIT D816V mutation. With poor response to conventional poly-chemotherapy, mast cell leukemia responded to the midostaurin treatment with a 50% overall response rate (ORR), but complete remission rate is approximately 0%. Therefore, the potential mechanisms of midostaurin resistance and the exact impacts of midostaurin on both gene expression profile and mast cell leukemia microenvironment in vivo are essential for design tailored combination therapy targeting both the tumor cells and the tumor microenvironment. Here we report a 59-year-old male mast cell leukemia patient with KIT F522C mutation treated with midostaurin. Single-cell sequencing of peripheral blood and whole exome sequencing (WES) of bone marrow were performed before and 10 months after midostaurin treatment. In accordance with the clinical response, compared to the pretreatment aberration, the decline of mast cells and increase of T-, NK, B-cells in peripheral blood, and the decrease of the KIT F522C mutation burden in bone marrow were observed. Meanwhile, the emergence of RUNX1 mutation, upregulations of genes expression (RPS27A, RPS6, UBA52, RACK1) on tumor cells, and increased frequencies of T and NK cells with TIGIT, CTLA4, and LAG3 expression were observed after midostaurin treatment, predicting the disease progression of this patient. As far as we know, this is the first case reporting the clinical, immunological, and molecular changes in mast cell leukemia patients before and after midostaurin treatment, illustrating the in vivo mechanisms of midostaurin resistance in mast cell leukemia, providing important clues to develop a sequential option to circumvent tumor progression after targeting oncogene addiction and prolong patients' survival.

Enhanced lipid metabolism confers the immunosuppressive tumor microenvironment in CD5-positive non-MYC/BCL2 double expressor lymphoma.

Lymphoma cells expressing CD5 (CD5+) confer inferior outcome of diffuse large B-cell lymphoma (DLBCL), especially in non-MYC/BCL2 double expressor (non-DE) patients. In tumor microenvironment, CD5+ non-DE tumor revealed increased proportion of immunosuppressive M2 macrophages and enhanced pathways related to macrophage activation and migration. In accordance to M2 activation, lipid metabolism was upregulated, including fatty acid uptake and fatty acid oxidation, which supplied energy for M2 macrophage polarization and activation. Meanwhile, CD36 expression was upregulated and strongly correlated to the proportion of M2 macrophages in CD5+ non-DE DLBCL. In vitro, a DLBCL cell line (LY10) overexpressing CD5 significantly increased M2 proportion in comparison with control when cocultured with peripheral blood mononuclear cells (PBMCs). The addition of metformin significantly decreased the M2 proportion and the CD36 expression level in the coculture systems, indicating that metformin could target altered lipid metabolism and decrease M2 macrophages in DLBCL, especially in CD5+ non-DE lymphoma. In conclusion, enhanced lipid metabolism and M2 macrophage activation contributed to the immunosuppressive tumor microenvironment and could be potential therapeutic targets in CD5+ non-DE DLBCL.

Bedaquiline-containing regimens in patients with pulmonary multidrug-resistant tuberculosis in China: focus on the safety.

BACKGROUND: World Health Organization recommends countries introducing new drug and short treatment regimen for drug resistant tuberculosis (DR-TB) should develop and implement a system for active pharmacovigilance that allows for detection, reporting and management of adverse events. The aim of the study is to evaluate the frequency and severity of adverse events (AEs) of bedaquiline-containing regimen in a cohort of Chinese patients with multidrug-resistant (MDR)/extensively drug-resistant (XDR)-TB based on active drug safety monitoring (aDSM) system of New Drug Introduction and Protection Program (NDIP). METHODS: AEs were prospectively collected with demographic, bacteriological, radiological and clinical data from 54 sites throughout China at patient enrollment and during treatment between February, 2018 and December, 2019. This is an interim analysis including patients who are still on treatment and those that have completed treatment. A descriptive analysis was performed on the patients evaluated in the cohort. RESULTS: By December 31, 2019, a total of 1162 patients received bedaquiline-containing anti-TB treatment. Overall, 1563 AEs were reported, 66.9% were classified as minor (Grade 1-2) and 33.1% as serious (Grade 3-5). The median duration of bedaquiline treatment was 167.0 [interquartile range (IQR): 75-169] days. 86 (7.4%) patients received 36-week prolonged treatment with bedaquiline. The incidence of AEs and serious AEs was 47.1% and 7.8%, respectively. The most frequently reported AEs were QT prolongation (24.7%) and hepatotoxicity (16.4%). There were 14 (1.2%) AEs leading to death. Out of patients with available corrected QT interval by Fridericia's formula (QTcF) data, 3.1% (32/1044) experienced a post-baseline QTcF ≥ 500 ms, and 15.7% (132/839) had at least one change of QTcF ≥ 60 ms from baseline. 49 (4.2%) patients had QT prolonged AEs leading to bedaquiline withdrawal. One hundred and ninety patients reported 361 AEs with hepatotoxicity ranking the second with high occurrence. Thirty-four patients reported 43 AEs of hepatic injury referred to bedaquiline, much lower than that referred to protionamide, pyrazinamide and para-aminosalicylic acid individually. CONCLUSIONS: Bedaquiline was generally well-tolerated with few safety concerns in this clinical patient population without any new safety signal identified. The mortality rate was generally low. These data inform significant positive effect to support the WHO recent recommendations for the wide use of bedaquiline.

[Identification of 23 unknown Li minority medicinal plants based on DNA barcoding].

DNA barcode molecular biological technique is used to identify the species of 23 unknown Li minority medicinal plants.DNA was extracted from 23 unknown medicines using the Plant Genomic DNA Extraction kit. The ITS2 and psbA-trnH regions were amplified and sequenced bi-directionally. The Codon Code Aligner V 7. 0. 1 was used to proofread and assemble the contigs and generated consensus sequences. All the sequences were submitted to Traditional Chinese Medicine DNA Barcode Database and NCBI Gen Bank to get information of the species identifications. If the maximum similarity of the identification result is ≥ 97%,exact species can be known. If it is between 97% and 90%,samples' genus can be confirmed; If it is <90%,then we can only confirm its family. Finally there are 17 samples can be identified to species level,5 can be identified to genus level and 1 can be identified to family level. This shows that DNA barcoding used in medicinal plants molecular identification,can identify unknown species rapidly and accurately.

Comparison of photosynthesis and fluorescent parameters between Dendrobium officinale and Dendrobium loddigesii.

OBJECTIVE: To investigate the photosynthesis and fluorescent parameters between Dendrobium officinale and Dendrobium loddigesii, based on which to provide helpful information for the artificial cultivation of these cultivars. METHODS: Seeds were placed on the MS medium supplemented with 0.2 mg/L NAA, 2% (w/v) sucrose, 15% (v/v) potato extracts and powered agar (pH 5.8). Two months after germination, seedlings (n = 10) were transferred onto rooting medium containing MS medium supplemented with 0.5 mg/L NAA, 3% (w/v) sucrose, 20% (v/v) potato extracts and 1‰ (w/v) activated carbon (pH 5.8) in a glass bottle (6.5 cm in diameter and 9.5 cm in height) with a white transparent plastic cap. Chlorophyll content was determined using the UV-Vis spectrophotometric method. In addition, rates of oxygen evolution and uptake were measured. The chlorophyll fluorescence was determined at room temperature using PAM 2000 chlorophyll fluorometer (Heinz Walz GmbH, Germany). RESULTS: From month 5 to month 10, the overall contents of both chlorophyll a and chlorophyll b were higher in D. loddigesii compared with those in D. officinale. No statistical differences were observed in the apparent photosynthetic rate (APR) between D. loddigesii and D. officinale. No statistical difference was noticed in the Fo, Fm and Fv between D. loddigesii and D. officinale (P > 0.05). Significant increase was noticed in the oxygen consuming in PSI in month-8 and month-10 compared with that of month-6 in D. loddigesii. Nevertheless, in the D. officinale, the oxygen consuming in PSI in month-6 was remarkably increased with those of month-8 and month-10, respectively. CONCLUSIONS: The photosynthesis and fluorescence parameters varied in the seedling of D. loddigesii and D. officinale. Such information could contribute to the artificial cultivation of these cultivars.

Electroacupuncture-induced dynamic processes of gene expression levels of endogenous opioid Peptide precursors and opioid receptors in the CNS of goats.

In order to investigate the dynamic processes of mRNA levels of proenkephalin, proopiomelanocortin, prodynorphin, and opioid receptors (δ-, µ-, and κ-receptor) induced by electroacupuncture (EA) in the central nerve system, goats were stimulated by EA of 60 Hz for 0.5 h at a set of Baihui, Santai, Ergen, and Sanyangluo points. The pain threshold was measured using the method of potassium iontophoresis. The mRNA levels of the three opioid peptide precursors and three opioid receptors were determined with quantitative real-time PCR and the levels of Met-enkephalin with SABC immunohistochemistry at 0.5 h before and at 0, 2, 4, 6, 8, 12, and 24 h after EA. The results showed that the pain threshold correlated (P < 0.01) with Met-enkephalin immunoactivities in the measured nuclei and areas of goats. The analgesic aftereffect lasted for 12 h at least. The mRNA levels of the three opioid peptide precursors and three opioid receptors began to increase at 0 h, reached the peak during the time from 4 h to 6 h or at 12 h, and remained higher at 24 h after EA was discontinued. These results suggested that the initiation of gene expression of opioid peptides and the three receptors may be associated with EA-induced analgesic aftereffect.

Effects of electroacupuncture of different frequencies on the release profile of endogenous opioid peptides in the central nerve system of goats.

To investigate the release profile of met-enkephalin, ß-endorphin, and dynorphin-A in ruminants' CNS, goats were stimulated by electroacupuncture of 0, 2, 40, 60, 80, or 100 Hz for 30 min. The pain threshold was measured using potassium iontophoresis. The peptide levels were determined with SABC immunohistochemisty. The results showed that 60 Hz increased pain threshold by 91%; its increasing rate was higher (P < 0.01) than any other frequency did. 2 Hz and 100 Hz increased met-enkephalin immunoactivities (P < 0.05) in nucleus accumbens, septal area, caudate nucleus, amygdala, paraventricular nucleus of hypothalamus, periaqueductal gray, dorsal raphe nucleus, and locus ceruleus. The two frequencies elicited ß-endorphin release (P < 0.05) in nucleus accumbens, septal area, supraoptic nucleus, ventromedial nucleus of hypothalamus, periaqueductal gray, dorsal raphe nucleus, locus ceruleus, solitary nucleus and amygdala. 60 Hz increased (P < 0.05) met-enkephalin or ß-endorphin immunoactivities in the nuclei and areas mentioned above, and habenular nucleus, substantia nigra, parabrachial nucleus, and nucleus raphe magnus. High frequencies increased dynorphin-A release (P < 0.05) in spinal cord dorsal horn and most analgesia-related nuclei. It suggested that 60 Hz induced the simultaneous release of the three peptides in extensive analgesia-related nuclei and areas of the CNS, which may be contributive to optimal analgesic effects and species variation.

[Effects of electroacupuncture of "Weizhong" (BL 40), "Sanyinjiao" (SP 6) and "Yinlingquan" (SP 9) on intravesical pressure and bladder adenosine triphosphate content in rabbits with acute urinary retention].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Weizhong" (BL 40), "Sanyinjiao" (SP 6) and "Yinlingquan" (SP 9) on changes of intravesical pressure and contents of adenosine triphosphate (ATP) in the urinary bladder tissue in rabbits with acute urine retention, so as to explore the relatively specificity of acupoints for urinary bladder problems. METHODS: Forty-eight male adult rabbits were randomly divided into normal control, model, Weizhong (BL 40, EA-BL 40), San-yinjiao (SP 6, EA-SP 6), Yinlingquan (SP9, EA-SP9) and non-acupoint (EA-NA) groups. Acute urinary retention model was established by filling the rabbits' bladder with normal saline at a volume of 2 times above their normal capacity for 2 hours. EA (2 Hz/15 Hz, 0.6 mA) was applied to bilateral "Weizhong" (BL 40), "Sanyinjiao" (SP 6) and "Yinlingquan" (SP 9) for 20 min, respectively. Intravesical pressure was detected by using a pressure transducer and an amplifier. Bladder ATP content was detected by using enzyme linked immunosorbent assay (ELISA). RESULTS: In comparison with pre-modeling, the intravesical pressure levels were decreased significantly after modeling in the model, EA-BL 40, EA-SP 6, EA-SP 9 and EA-NA groups (P < 0.05). Compared with pre-EA, intravesical pressure values were increased remarkably in the EA-BL 40, EA-SP 6, EA-SP 9 and EA-NA groups (P < 0.05). The percentages of the increased vesical pressure after EA were considerably higher in the EA-BL 40, EA-SP 6, EA-SP 9 and EA-NA groups than in the model group (P < 0.05). Compared with the normal group, bladder ATP content in the model group was reduced significantly (P < 0.05); while in comparison with the model group, ATP contents in the EA-BL 40, EA-SP 6 and EA-SP 9 groups were up-regulated apparently (P < 0.05). The bladder ATP level of the EA-BL 40 group was significantly higher than those of EA-SP 6 and EA-SP 9 groups (P < 0.05). No significant differences were found between the model and EA-NA groups, and between the EA-SP6 and EA-SP9 groups in bladder ATP contents (P > 0.05), and among the EA-BL 40, EA-SP 6 and EA-SP 9 groups in the percentages of the increased intravesical pressure (P > 0.05). CONCLUSION: EA of BL 40, SP 6 and SP 9 can significantly raise intravesical pressure and bladder ATP content in urine retention rabbits, which may contribute to its effect in improving urinary retention.

Y4lO of Rhizobium sp. strain NGR234 is a symbiotic determinant required for symbiosome differentiation.

Type 3 (T3) effector proteins, secreted by nitrogen-fixing rhizobia with a bacterial T3 secretion system, affect the nodulation of certain host legumes. The open reading frame y4lO of Rhizobium sp. strain NGR234 encodes a protein with sequence similarities to T3 effectors from pathogenic bacteria (the YopJ effector family). Transcription studies showed that the promoter activity of y4lO depended on the transcriptional activator TtsI. Recombinant Y4lO protein expressed in Escherichia coli did not acetylate two representative mitogen-activated protein kinase kinases (human MKK6 and MKK1 from Medicago truncatula), indicating that YopJ-like proteins differ with respect to their substrate specificities. The y4lO gene was mutated in NGR234 (strain NGROmegay4lO) and in NGR Omega nopL, a mutant that does not produce the T3 effector NopL (strain NGR Omega nopLOmegay4lO). When used as inoculants, the symbiotic properties of the mutants differed. Tephrosia vogelii, Phaseolus vulgaris cv. Yudou No. 1, and Vigna unguiculata cv. Sui Qing Dou Jiao formed pink effective nodules with NGR234 and NGR Omega nopL Omega y4lO. Nodules induced by NGR Omega y4lO were first pink but rapidly turned greenish (ineffective nodules), indicating premature senescence. An ultrastructural analysis of the nodules induced by NGR Omega y4lO revealed abnormal formation of enlarged infection droplets in ineffective nodules, whereas symbiosomes harboring a single bacteroid were frequently observed in effective nodules induced by NGR234 or NGR Omega nopL Omega y4lO. It is concluded that Y4lO is a symbiotic determinant involved in the differentiation of symbiosomes. Y4lO mitigated senescence-inducing effects caused by the T3 effector NopL, suggesting synergistic effects for Y4lO and NopL in nitrogen-fixing nodules.

Study on the identification of HIV/AIDS and its surveillance system in rural areas of central China / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore the effective strategies for human immunodeficiency virus (HIV) surveillance and identification in rural areas of central China.</p><p><b>METHODS</b>In a selected rural prefecture area of central China, an epidemiological investigation was conducted for all reported HIV/AIDS cases. A historical and analytic review was performed, with particular interests in examining the HIV epidemic reporting and identification system.</p><p><b>RESULTS</b>Among all 626 reported HIV-infected individuals in the selected prefecture, 86.1% (539/626) of them were infected through commercial plasma donation or clinically operated blood transfusion. With respect to disease surveillance and identification in the area, 52.2% (327/626) of all the cases were reported by hospitals or clinics. The number and proportion of HIV/AIDS cases identified or reported at county, prefecture, and provincial levels were 207 (33.1%), 303 (48.4%) and 116 (18.5%), respectively. The number and proportion of HIV/AIDS identified through specific epidemiological investigations and/or voluntary testing had been increased in recent years. In addition, among HIV/AIDS cases that were clinically identified, the proportion of those who were identified as outpatients had steadily increased,with the highest proportion (59.3%) observed in 2004.</p><p><b>CONCLUSIONS</b>In rural areas of central China where the major mode of HIV transmission was through commercial plasma donation or clinically operated blood transfusion, hospitals and clinics seemed to have played and would continue to play important roles regarding HIV identification and surveillance. The role of institutions or settings at the prefecture level regarding HIV identification and surveillance should not be ignored.</p>

Computed tomographic assessment of the surgical risks associated with fibrocavernous pulmonary tuberculosis.

PURPOSE: We evaluated the surgical risks associated with fibrocavernous pulmonary tuberculosis by retrospectively examining chest computed tomography (CT) scans. METHODS: We reviewed the records of 40 patients who underwent pulmonary resection for fibrocavernous pulmonary tuberculosis, for whom preoperative CT scans were available. The disease was categorized as class I, defined as a cavity within one lobe without remarkable pleural thickness, in 21 patients; class II, defined as a cavity extending beyond one lobe or within one lobe with remarkable pleural thickness, in 10 patients; and class III, defined as bilateral cavities, in 9 patients. Four of the nine patients with bilateral cavities underwent bilateral pulmonary resection and five underwent unilateral pulmonary resection. The study parameters were intraoperative blood loss, operative time, hospital stay, major operative morbidity, and hospital death. RESULTS: Intraoperative blood loss and operative time were significantly greater and hospital stay was significantly longer in patients with advanced disease (P = 0.046, P = 0.000, and P = 0.143, respectively). Major surgical morbidity mainly occurred in association with advanced disease (P = 0.028) at the following incidences: class I, 5%; class II, 30%; class III, 44.4%. Two hospital deaths occurred, both following bilateral pulmonary resection for class III disease, accounting for an overall 5% mortality rate. CONCLUSION: The surgical risks associated with fibrocavernous pulmonary tuberculosis were well correlated with anatomic involvement, according to the extent of cavitation and the severity of pleural thickness, as depicted by CT. Staged pulmonary resection or the combination of one-sided resection with other modalities is recommended for the treatment of bilateral cavities.

[Non-osteogenic malocclusion after anatomic reduction and miniplate rigid fixation in condylar neck and subcondylar fractures].

OBJECTIVE: To analysis the cause of malocclusion that was found in patients suffering from condylar neck and subcondylar fractures after perfectly anatomic reduction and rigid fixation. METHODS: There were 23 cases of malocclusion in the patients of condylar neck and subcondylar fractures after anatomic reduction and rigid fixation during 1994 to 2001. The possible reasons were studied, and the treatment methods were proposed. RESULTS: Both distortion and(or) fracture of miniplates and displacement anain of the condylar process after operation were not found in all 23 cases. There were 6 cases of open bite at the molar teeth of the fractured side, while the normal occlusions were showed in the nonfractured side. 11 cases showed that the whole mandibles were moved slightly to the nonfractured side. 4 cases showed that the mandible of fractured side moved slightly to the nonfractured side and the occlusion of nonfractured side were normal. 2 cases showed premature contact at the fractured molar teeth and the anterior teeth showed open bite. There were 5 cases of facial nerve damages, 2 cases wound infection, 2 cases TMJ chronic pain and 1 cases TMJ click and(or) murmur. CONCLUSIONS: Although the anatomical reduction and miniplate rigid fixation were operated on the condylar neck and subcondylar fractures, there were probably having the chance of showing malocclusion. The malocclusion was not caused by the fracture itself, but in the TMJ and/or muscular injury. Therefore, the malocclusion can be cured with intermaxillary elastic traction and functional exercise. Sometimes, the occlusal adjustment may be performed to resume the normal occlusion. In any case, operation is not needed.

Treatment of lumbar disk herniation with MRI guidance injection of oxygen-ozone / 中华放射学杂志

Objective To evaluate the applied value of percutaneous oxygen-ozone injection in the treatment of lumbar disc herniation under open 0.23 T MRI guidance.Methods Mounted with ipath 200 optical tracking system,MR-guided injection of oxygen-ozone were performed via a medial border of the articular processes approach in 73 patients with clinically diagnosed LDH.MR compatible 19.5G or 21.0 G biopsy needle was used. Discography was performed in order to select indication before injection oxygen- ozone into nucleus pulposus in 26 patients.Sixty-four patients were injected to three sites:(1)Six to 10 ml oxygen-ozone was injected into discs centers,injected and suctioned alternately in order to make nucleus pulposus oxidation thoroughly.(2)The needle was withdrawn according to the scale of biopsy needle and optical tracking.Then,10 ml oxygen-ozone was injected into disc herniation. (3)After that,needle was withdrawn further about 1.0—1.5 cm to outside of annulus fibrosus.Fifteen to 20 ml oxygen-ozone was injected into intervertebral foramina around nerve roots.The oxygen-ozone concentration was 35—45?g/ml. Nine patients were only performed injection of oxygen-ozone into around nerve root,while not injection oxygen-ozone to nucleus pulposus for considering bad curative effect after discography.Results All of 73 patients were successfully local targeted and treated under MRI guidance without serious complications, such as nerve root injury.After 3—6 months follow-up,total overall efficacy was 91.3% with the excellent in 28,good in 39,and poor in 6,respectively.Conclusion Open MR-guided injection of oxygen-ozone, mounted with optical tracking system,is a safe and effective minimally invasive therapy for treating LDH.