The SDS22:PP1:I3 complex: SDS22 binding to PP1 loosens the active site metal to prime metal exchange.

SDS22 and Inhibitor-3 (I3) are two ancient regulators of protein phosphatase 1 (PP1) that regulate multiple essential biological processes. Both SDS22 and I3 form stable dimeric complexes with PP1; however, and atypically for PP1 regulators, they also form a triple complex, where both proteins bind to PP1 simultaneously (SPI complex). Here we report the crystal structure of the SPI complex. While both regulators bind PP1 in conformations identical to those observed in their individual PP1 complexes, PP1 adopts the SDS22-bound conformation, which lacks its M1 metal. Unexpectedly, surface plasmon resonance (SPR) revealed that the affinity of I3 for the SDS22:PP1 complex is ∼10-fold lower than PP1 alone. We show that this change in binding affinity is solely due to the interaction of I3 with the PP1 active site, specifically PP1's M2 metal, demonstrating that SDS22 likely allows for PP1 M2 metal exchange and thus PP1 biogenesis.

Molecular basis of ß-lactam antibiotic resistance of ESKAPE bacterium E. faecium Penicillin Binding Protein PBP5.

Penicillin-binding proteins (PBPs) are essential for the formation of the bacterial cell wall. They are also the targets of ß-lactam antibiotics. In Enterococcus faecium, high levels of resistance to ß-lactams are associated with the expression of PBP5, with higher levels of resistance associated with distinct PBP5 variants. To define the molecular mechanism of PBP5-mediated resistance we leveraged biomolecular NMR spectroscopy of PBP5 - due to its size (>70 kDa) a challenging NMR target. Our data show that resistant PBP5 variants show significantly increased dynamics either alone or upon formation of the acyl-enzyme inhibitor complex. Furthermore, these variants also exhibit increased acyl-enzyme hydrolysis. Thus, reducing sidechain bulkiness and expanding surface loops results in increased dynamics that facilitates acyl-enzyme hydrolysis and, via increased ß-lactam antibiotic turnover, facilitates ß-lactam resistance. Together, these data provide the molecular basis of resistance of clinical E. faecium PBP5 variants, results that are likely applicable to the PBP family.

The effect of microRNA-663b in the inhibition of interleukin-1-induced nucleus pulposus cell apoptosis and inflammatory response.

The aim of this study was to explore the role and pathological mechanism of microRNA-663b in interleukin-1beta (IL-1ß)-induced inflammation and apoptosis of nucleus pulposus cells. First, the best concentration and time to construct the nucleus pulposus cell inflammation model was screen out. Overexpression or inhibition of miR-663b expression was performed by adding microRNA-663b mimic or microRNA-663b inhibitor. 293T cells were transfected according to experimental requirements. The luciferase activity of each group was detected to analyze the targeted regulation of microRNA-663b on interleukin-1 receptor (IL1R1). Compared with the mimic negative control (NC) group, the expression of inflammatory factors in the microRNA-663b overexpression group was inhibited (P<0.05), and the expression of type 2 collagen and polysaccharide protein increased (P<0.05), and the apoptosis of nucleus pulposus cells was inhibited (P<0.01), and the number of TUNEL-positive cells decreased significantly (P<0.01), and the microRNA and protein expression of IL1R1, the ratio of P-P65/P65 and phospho-nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (P-IκBα)/nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (IκBα) protein expression were significantly decreased (P<0.05). The expression of inflammatory factors in the miR-663b inhibitor group was significantly higher than that in the inhibitor NC group (P<0.01), and the expression of type 2 collagen and polysaccharide protein was significantly decreased (P<0.01), and the number of apoptosis cells and TUNEL staining positive cells increased (p<0.01). The expression of IL1R1 gene and protein was significantly increased (P<0.01). The ratio of P-P65/P65 and P-IκBα/IκBα protein expression increased (P<0.05). IL1R1 is a downstream target gene of microRNA-663b. MicroRNA-663b may down-regulate the expression of IL1R1 at the transcriptional level by targeting IL1R1, inhibit the inflammatory response of nucleus pulposus cells, and slow down the degeneration of nucleus pulposus cells.

[Principle and routine operation of laser assisted periodontal surgery].

Periodontitis is one of the most common infectious oral diseases, which can cause destruction of periodontal supporting tissues and even tooth mobility and loss. Controlling infection, eliminating inflammation, restoring the physiological shape of periodontal tissues, and meeting functional and aesthetic needs are the main goals of periodontal treatment. When periodontitis develops to a more severe stage, surgical treatment is necessary to handle soft and hard tissues for good treatment results. Since the development of the first Nd:YAG laser dedicated to dental medicine by Myers in 1990, over 30 years of clinical and basic research have shown that lasers have tremendous potential in assisting periodontal surgery. This article summarizes the principles and operational routines of laser-assisted periodontal surgery, aiming to provide clinical reference for diagnosis and treatment.

[Contrast-enhanced ultrasonography with intra-glandular contrast injection can improve the diagnostic accuracy of central compartment lymph node metastasis of thyroid cancer].

OBJECTIVE: To investigate the value of lymphatic contrast-enhanced ultrasound (LCEUS) with intra-glandular injection of contrast agent for diagnosis of central compartment lymph node metastasis of thyroid cancer. METHODS: From November, 2020 to May, 2022, the patients suspected of having thyroid cancer and scheduled for biopsy at our center received both conventional ultrasound and LCEUS examinations of the central compartment lymph nodes before surgery. All the patients underwent surgical dissection of the lymph nodes. The perfusion features in LCEUS were classified as homogeneous enhancement, heterogeneous enhancement, regular/irregular ring, and non-enhancement. With pathological results as the gold standard, we compared the diagnostic ability of conventional ultrasound and LCEUS for identifying metastasis in the central compartment lymph nodes. RESULTS: Forty-nine patients with 60 lymph nodes were included in the final analysis. Pathological examination reported metastasis in 34 of the lymph nodes, and 26 were benign lymph nodes. With ultrasound findings of heterogeneous enhancement, irregular ring and non-enhancement as the criteria for malignant lesions, LCEUS had a diagnostic sensitivity, specificity and accuracy of 97.06%, 92.31% and 95% for diagnosing metastatic lymph nodes, respectively, demonstrating its better performance than conventional ultrasound (P < 0.001). Receiver-operating characteristic curve analysis showed that LCEUS had a significantly greater area under the curve than conventional ultrasound for diagnosing metastatic lymph nodes (94.7% [0.856-0.988] vs 78.2% [0.656-0.878], P=0.003). CONCLUSION: LCEUS can enhance the display and improve the diagnostic accuracy of the central compartment lymph nodes to provide important clinical evidence for making clinical decisions on treatment of thyroid cancer.

Inhibitor-3 inhibits Protein Phosphatase 1 via a metal binding dynamic protein-protein interaction.

To achieve substrate specificity, protein phosphate 1 (PP1) forms holoenzymes with hundreds of regulatory and inhibitory proteins. Inhibitor-3 (I3) is an ancient inhibitor of PP1 with putative roles in PP1 maturation and the regulation of PP1 activity. Here, we show that I3 residues 27-68 are necessary and sufficient for PP1 binding and inhibition. In addition to a canonical RVxF motif, which is shared by nearly all PP1 regulators and inhibitors, and a non-canonical SILK motif, I3 also binds PP1 via multiple basic residues that bind directly in the PP1 acidic substrate binding groove, an interaction that provides a blueprint for how substrates bind this groove for dephosphorylation. Unexpectedly, this interaction positions a CCC (cys-cys-cys) motif to bind directly across the PP1 active site. Using biophysical and inhibition assays, we show that the I3 CCC motif binds and inhibits PP1 in an unexpected dynamic, fuzzy manner, via transient engagement of the PP1 active site metals. Together, these data not only provide fundamental insights into the mechanisms by which IDP protein regulators of PP1 achieve inhibition, but also shows that fuzzy interactions between IDPs and their folded binding partners, in addition to enhancing binding affinity, can also directly regulate enzyme activity.

[A clinicopathological classification of space-occupying lesions of the orbit in 1 913 patients from 2000 to 2021].

Objective: To investigate the histopathological classification of orbital space-occupying lesions. Methods: This is a retrospective case series study. The clinical and pathological data of 1 913 tissue specimens from 1 913 patients with space-occupying lesions of the orbit which were examined in the Second Affiliated Hospital, Zhejiang University School of Medicine from January 2000 to December 2021 were collected. The mass lesions were classified based on histogenesis, pathological nature and age. Results: There were 913 males (47.7%) and 1 000 females (52.3%). The lesions were benign in 1 489 patients (77.8%) and malignant in 424 patients (22.2%). Based on histogenesis, there were 521 vasculogenic lesions (27.2%), which rancked first, 407 cystoid lesions (21.3%), 277 lymphoproliferative lesions (14.5%), 182 lacrimal gland lesions (9.5%) and 121 inflammatory lesions (6.3%). By pathological nature, there were 1 489 benign lesions, including cavernous hemangioma (275, 14.4%), dermoid cyst (225, 11.8%), other hemangiomas (199, 10.4%), epidermoid cyst (136, 7.1%) and benign mixed tumor of the lacrimal gland (134, 7.0%), and 257 malignant lesions, including lymphoma (210, 11.0%) and sebaceous gland carcinoma (47, 2.5%). The age of all patients ranged from 0 to 90 years, while 247 lesions (12.9%) occurred in patients aged 0 to18 years, 1 270 lesions (66.4%) in patients aged 19 to 59 years, and 396 lesions (20.7%) in patients aged 60 to 90 years. Conclusions: In 22 years, almost 2/3 benign orbital lesions in the Second Affiliated Hospital, Zhejiang University School of Medicine occurred in young and middle-aged patients, and males were fewer than females. The most common benign orbital tumors was cavernous hemangioma, followed by dermoid cyst and epidermoid cyst. And the most common malignant orbital tumor was lymphoma, which occurred more frequently in older patients.

Postoperative Complications from Breast and Axillary Surgery.

Although surgery of the breast and axilla is generally well-tolerated by patients, the breast surgeon recognizes that complications can occur even when operating with experience on the lowest risk patients. The operative repertoire ranges from breast conserving surgery, mastectomy (including skin-sparing and nipple-sparing types), to modified radical mastectomy, with each procedure carrying a different expected surgical morbidity. Patients and families who are fully informed of potential complications before their operation describe greater trust in their surgeon and are better able to co-manage complications with the surgical team, when they occur.

[Clinical application of complete laparoscopic gastrectomy with function preservation of cardia in gastric carcinoma].

To investigate the feasibility and safety of total laparoscopic cardia function preserving gastrectomy for gastric carcinoma. Clinical data of 10 patients undergoing total laparoscopic cardia function preserving gastrectomy for gastric carcinoma from November 2020 to December 2021 were retrospectively collected. There were 7 males and 3 females. The mean age was (66.1±12.9) years (ranged from 38 to 86 years). All of the 10 patients were successfully performed total laparoscopic cardia function preserving gastrectomy without conversion to laparotomy. The time of digestive tract reconstruction was (24.8±3.3) min (20-30 min), and the intraoperative blood loss was (35±24) ml(20-100 ml). The time of postoperative exhaust was (2.5±0.9) days(2-3 d), the time of postoperative liquid diet was (2.25±0.87) days(2-3 d), postoperative hospital stay was (9.5±2.1) days(6-13 d). No surgical complications such as bleeding, anastomotic fistula or anastomotic stenosis occurred. Postoperative pathology showed that the proximal and distal margins of resected specimens were negative. Patients were followed up for 2 to 15 months, respectively. No death or tumor recurrence and metastasis occurred during the follow-up period. There were no symptoms of reflux after operation. Compared with total gastrectomy and proximal gastrectomy, total laparoscopic cardia function preserving gastrectomy can theoretically reduce the incidence of reflux esophagitis. We used manual suture method for digestive tract reconstruction, which can reduce the application of 2-3 stapling studs and reduce the cost of surgical materials. Compared with subtotal gastrectomy, total laparoscopic cardia function preserving gastrectomy has the advantages of more thorough lymph node dissection, with little residual gastric tissue; therefore, the blood supply is relatively better. The incidence of reflux esophagitis of total laparoscopic cardia function preserving gastrectomy for gastric cancer may was lower than total gastrectomy.

Ultrasound for the detection of the pyramidal lobe of the thyroid gland.

PURPOSE: The pyramidal lobe (PL) is an ancillary lobe of the thyroid gland that can be affected by the same pathologies as the rest of the gland. We aimed to assess the diagnostic performance of high-resolution sonography in the detection of the PL with verification by dissection and histological examination. METHODS: In a prospective, cross-sectional mono-center study, 50 fresh, non-embalmed cadavers were included. Blinded ultrasound examination was performed to detect the PL by two investigators of different experience levels. If the PL was detected with ultrasound, dissection was performed to expose the PL and obtain a tissue sample. When no PL was detected with ultrasound, a tissue block of the anterior cervical region was excised. An endocrine pathologist microscopically examined all tissue samples and tissue blocks for the presence of thyroid parenchyma. RESULTS: The prevalence of the PL was 80% [40/50; 95% CI (68.9%; 91.1%)]. Diagnostic performance for both examiners was: sensitivity (85.0%; 42.5%), specificity (50.0%; 60.0%), positive predictive value (87.2%; 81.0%), negative predictive value (45.5%; 21.0%) and accuracy (78.0%; 46.0%). Regression analysis demonstrated that neither thyroid parenchyma echogenicity, thyroid gland volume, age nor body size proved to be covariates in the accurate detection of a PL (p > .05). CONCLUSION: We report that high-resolution ultrasound is an adequate examination modality to detect the PL. Our findings indicate a higher prevalence than previously reported. Therefore, the PL may be regarded as a regular part of the thyroid gland. We also advocate a dedicated assessment of the PL in routine thyroid ultrasound.

[Clinical effect of minimally invasive duodenum preserving pancreatic head resection for benign and pre-malignant lesions of pancreatic head].

Objective: To examine the clinical effect of minimally invasive duodenum preserving pancreatic head resection(DPPHR) for benign and pre-malignant lesions of pancreatic head. Methods: The clinical data of patients with diagnosis of benign or pre-malignant pancreatic head tumor were retrospectively collected and analyzed,all of them underwent laparoscopic or robotic DPPHR between October 2015 and September 2021 at Division of Gastrointestinal and Pancreatic surgery,Zhejiang Provincial People's Hospital. Thirty-three patients were enrolled with 10 males and 23 females. The age(M(IQR)) was 54(32) years old(range: 11 to 77 years old) and the body mass index was 21.9(2.9)kg/m2(range: 18.1 to 30.1 kg/m2). The presenting symptoms included abdominal pain(n=12), Whipple triad(n=2), and asymptomatic(n=19). There were 7 patients with hypertension and 1 patient with diabetes mellitus. There were 19 patients who were diagnosed as American Society of Anesthesiologists class Ⅰ and 14 patients who were diagnosed as class Ⅱ. The student t test,U test, χ2 test or Fisher exact test was used to compare continuous data or categorized data,respectively. All the perioperative data and metabolic morbidity were analyzed and experiences on minimally invasive DPPHR were concluded. Results: Fourteen patients underwent laparoscopic DPPHR,while the rest of 19 patients received robotic DPPHR. Indocyanine green fluorescence imaging was used in 19 patients to guide operation. Five patients were performed pancreatico-gastrostomy and the rest 28 patients underwent pancreaticojejunostomy. Pathological outcomes confirmed 9 solid pseudo-papillary neoplasms, 9 intraductal papillary mucinous neoplasms, 7 serous cystic neoplasms, 6 pancreatic neuroendocrine tumors, 1 mucous cystic neoplasm, 1 chronic pancreatitis. The operative time was (309.4±50.3) minutes(range:180 to 420 minutes),and the blood loss was (97.9±48.3)ml(range:20 to 200 ml). Eighteen patients suffered from postoperative complications,including 3 patients experienced severe complications(Clavien-Dindo Grade ≥Ⅲ). Pancreatic fistula occurred in 16 patients,including 8 patients with biochemical leak,7 patients with grade B pancreatic fistula and 1 patient with grade C pancreatic fistula. No one suffered from the duodenal necrosis and none perioperative death was occurred. The length of hospital stay was 14(7) days (range:6 to 87 days). The follow-up was 22.6(24.5)months(range:2 to 74 months). None suffered from recurrence or metastasis. During the follow-up,all the patients were free of refractory cholangitis. Moreover,in the term of endocrine dysfunction,no postoperative new onset of diabetes mellitus were observed in the long-term follow-up. However,in the view of exocrine insufficiency,pancreatic exocrine insufficiency and non-alcoholic fatty liver disease (NAFLD) was complicated in 2 and 1 patient,respectively,with the supplement of pancreatic enzyme,steatorrhea and weight loss relieved,but NAFLD was awaited to be seen. Conclusions: Minimally invasive DPPHR is feasible and safe for benign or pre-malignant lesions of pancreatic head. Moreover,it is oncological equivalent to pancreaticoduodenectomy with preservation of metabolic function without refractory cholangitis.

The 2021 ultrafast spectroscopic probes of condensed matter roadmap.

In the 60 years since the invention of the laser, the scientific community has developed numerous fields of research based on these bright, coherent light sources, including the areas of imaging, spectroscopy, materials processing and communications. Ultrafast spectroscopy and imaging techniques are at the forefront of research into the light-matter interaction at the shortest times accessible to experiments, ranging from a few attoseconds to nanoseconds. Light pulses provide a crucial probe of the dynamical motion of charges, spins, and atoms on picosecond, femtosecond, and down to attosecond timescales, none of which are accessible even with the fastest electronic devices. Furthermore, strong light pulses can drive materials into unusual phases, with exotic properties. In this roadmap we describe the current state-of-the-art in experimental and theoretical studies of condensed matter using ultrafast probes. In each contribution, the authors also use their extensive knowledge to highlight challenges and predict future trends.

Analysis of neurotransmitters associated with neuropsychiatric status in workers following lead exposure.

OBJECTIVE: To explore the correlation between neuropsychiatric status and blood neurotransmitter in lead workers, and to provide theoretical basis for the prevention and treatment of lead workers. SUBJECTS AND METHODS: The study applied cross-sectional survey, 74 occupational lead exposed workers in a battery factory in a city of Hebei province were selected as the lead exposed group, and 62 workers (non-lead workers) were selected as the control group. The occupational health symptoms questionnaire and health examination and POMS (Profile of Mood State, POMS) emotional test questionnaire were applied to investigate the nearly emotional status of the studied objects, ICP-MS was used to determine the blood lead level of all subjects, HPLC (High performance liquid chromatography, HPLC) was applied to determine the concentration of neurotransmitter in peripheral blood of all studied subjects, and all results were applied the Pearson's correlation analysis. RESULTS: The blood lead concentration of the lead workers group (163.23±40.77 ug/L) was significantly higher than that in the control group (43.62±14.50 ug/L), and the difference was statistically significant. From the analysis of neuropsychiatric status, the neurological symptoms in the lead workers group were higher than that in the control group, among which the symptoms of sleep disturbance, dizziness, fatigue, numbness of limbs and dampness and coldness of limbs were more obvious. Among the symptoms of digestive system, the incidence of abdominal pain, abdominal distension, constipation and nausea and vomiting were higher. According to the POMS emotion questionnaire, the scores of 5 negative emotions and 1 positive emotion in the lead exposure group were higher than that in the control group, and the difference was statistically significant. Related to the control group, the concentration of neurotransmitters such as DA, 5-HT, GABA, Gly, Trp and Glu were statistically decreased, p<0.001. There was a negative correlation between neurotransmitters in peripheral blood and blood lead levels in lead workers, among which 5-HT had the greatest correlation with lead levels (r=-0.569, p<0.001). 5-HT and Trp were significantly correlated with tension-anxiety (T), depression-depression (D), anger-hostility (A), Vigor-hyperactivity (V), fatigue-inertia (F), and confusion-confusion (C). 5-HT, Trp and GABA were significantly correlated with the survey symptoms, among which, the sleep disorder, constipation and fatigue had most significantly positive correlation with 5-HT or Trp, r-value was respectively 0.373, 0.233 and 0.563. CONCLUSIONS: Lead exposure not only causes the alteration of neuropsychiatric behavior of lead workers, but also changes gastrointestinal symptoms. Serotonin may be involved as the main neurotransmitter synthesized in intestinal, and the synthesis and metabolism may be regulated by intestinal flora.

Recent advances on TMDCs for medical diagnosis.

Transition metal dichalcogenides (TMDCs), such as MoS2 and WS2, have attracted much attention in biosensing and bioimaging due to its excellent stability, biocompatibility, high specific surface area, and wide varieties. In this review, we overviewed the application of TMDCs in biosensing and bioimaging. Firstly, the synthesis methods and surface functionalization methods of TMDCs were summarized. Secondly, according to the working mechanism, we classified and gave a detailed account of the latest research progress of TMDC-based biosensing for the detection of the enzyme, DNA, and other biological molecules. Then, we outlined the recent progress of applying TMDCs in bio-imaging, including fluorescence, X-ray computed tomographic, magnetic response imaging, photographic and multimodal imaging, respectively. Finally, we discussed the future challenges and development direction of the application of TMDCs in medical diagnosis. Also, we put forward our view on the opportunity of TMDCs in the big data of modern medical diagnosis.

Brain-specific monoallelic expression of bovine UBE3A is associated with genomic position.

Genomic imprinting is a rare epigenetic process in mammalian cells that leads to monoallelic expression of a gene with a parent-specific pattern. The UBE3A (ubiquitin protein ligase E3A) gene is imprinted with maternal allelic expression in the brain but biallelically expressed in all other tissues in humans. The silencing of the paternal UBE3A allele is thought to be caused by the paternally expressed antisense RNA transcript of UBE3A-ATS. The aberrant imprinted expression of the UBE3A is associated with several neurodevelopmental syndromes and psychological disorders. Cattle are a valuable model species in determining the genetic etiology of sporadic human disorder, and maternal expression of UEB3A has been revealed by next-generation sequencing study in the bovine conceptus. In this study, we investigated the allelic expression of UBE3A and UBE3A-ATS in adult bovine somatic tissues. To confirm the splicing pattern of bovine UBE3A, five 5' alternative transcripts (MT210534-MT210538) were first obtained from bovine brain tissue by RT-PCR. Based on 10 SNP genotypes, we found that the brain-specific monoallelic expression of bovine UBE3A did not occur along the entire locus, and there was a shift from biallelic expression to monoallelic expression in exon 14 of the UBE3A gene. However, the brain-specific monoallelic expression of bovine UBE3A-ATS occurred in the entire gene. These observations demonstrated that the monoallelic expression did not occur along the bovine UBE3A entire locus and was associated with the genomic position.

Serum expression of EAAT2 and ADORA2A in patients with different degrees of Alzheimer's disease.

OBJECTIVE: This study aimed to explore the correlation between serum EAAT2 and ADORA2A levels and Alzheimer's disease (AD). PATIENTS AND METHODS: A total of 68 patients with AD treated in our hospital from April 2017 to January 2019 were enrolled and assigned to group A, and 60 healthy individuals undergoing physical examinations in the same period were enrolled and assigned to group B. Enzyme-linked immunosorbent assay (ELISA) was used to measure the expression of serum EAAT2 and ADORA2A in the two groups, receiver operating characteristic (ROC) curve to assess the predictive value of diagnostic efficacy, Spearman correlation to perform correlation analysis, and multivariate logistic analysis to analyze risk factors of prognosis. RESULTS: Patients from group A showed significantly higher serum ADORA2A level and lower serum EAAT2 level than individuals from group B (all p<0.001). The severity of AD was negatively correlated with the relative expression of serum EAAT2 (r=-0.7286, p<0.001), positively correlated with the relative expression of serum ADORA2A (r=0.7381, p<0.001). The sensitivity, specificity, and area under the curve (AUC) of EAAT2 alone for the diagnosis of AD were 85.00%, 82.35%, and 0.8853, respectively, and those of ADORA2A alone for the diagnosis of AD were 71.67%, 79.41.00%, and 0.8369, respectively. Univariate and multivariate Logistic regression analysis showed that disease severity, EAAT2, and ADORA2A were independent risk factors of the prognosis of AD. CONCLUSIONS: Patients with AD have highly expressed ADORA2A and lowly expressed EAAT2 in the serum. EAAT2 and ADORA2A may play parts in the progression of AD, and they can act as potential serum biomarkers for the diagnosis and disease assessment of AD.

Structural Optimization of 2,3-Dihydro-1H-cyclopenta[b]quinolines Targeting the Noncatalytic RVxF Site of Protein Phosphatase 1 for HIV-1 Inhibition.

Combination antiretroviral therapy (cART) suppresses human immunodeficiency virus-1 (HIV-1) replication but is unable to permanently eradicate HIV-1. Importantly, cART does not target HIV-1 transcription, which is reactivated in latently infected reservoirs, leading to HIV-1 pathogenesis including non-infectious lung, cardiovascular, kidney, and neurodegenerative diseases. To address the limitations of cART and to prevent HIV-1-related pathogenesis, we developed small molecules to target the noncatalytic RVxF-accommodating site of protein phosphatase-1 (PP1) to prevent HIV-1 transcription activation. The PP1 RVxF-accommodating site is critical for the recruitment of regulatory and substrate proteins to PP1. Here, we confirm that our previously developed 1E7-03 compound binds to the PP1 RVxF-accommodating site. Iterative chemical alterations to 1E7-03 furnished a new analogue, HU-1a, with enhanced HIV-1 inhibitory activity and improved metabolic stability compared to 1E7-03. In a Split NanoBit competition assay, HU-1a primarily bound to the PP1 RVxF-accommodating site. In conclusion, our study identified HU-1a as a promising HIV-1 transcription inhibitor and showed that the PP1 RVxF-accommodating site is a potential drug target for the development of novel HIV-1 transcription inhibitors.

[Application value of non-invasive ventilation combined with high flow nasal cannula oxygen therapy in sequential treatment of patients with chronic obstructive pulmonary disease after mechanical ventilation].

Objective: To investigate the value of non-invasive ventilation (NIV) combined with high flow nasal cannula oxygen therapy (HFNCO) in sequential treatment of patients with chronic obstructive pulmonary disease after mechanical ventilation. Methods: Chronic obstructive pulmonary disease with acute exacerbation (AECOPD) patients with invasive mechanical ventilation (MV) and successful withdrawal admitted into Huxi Affiliated Hospital of Jining Medical College from January 2018 to December 2019 were enrolled for perspective study. The patients were divided into treatment group (n=40) and control group (n=33) by random number table method. The treatment group was given NIV and HFNCO, the control group was given NIV treatment alone. Bedside ultrasound was used to measure the patients' diaphragmatic motion, and the differences between the two groups of patients before treatment, 24, 48 and 72 h after treatment were compared in diaphragmatic excursions during quiet breathing (DEq), diaphragmatic excursions during deep breathing(DEd), diaphragmatic shallow fast breathing index (D-RSBI), arterial oxygen partial pressure (PaO(2)), arterial partial pressure of carbon dioxide (PaCO(2)), re-tracheal intubation rate, mortality rate for 28 days and average duration of NPPV treatment within 3 days. Results: There were no statistically significant differences in DEq, DEd, D-RSBI, PaO(2) and PaCO(2) between the two groups before treatment (all P>0.05). After 24 h treatment, DEd decreased in both groups, D-RSBI increased in both groups, However, D-RSBI [(1.33±0.56) vs (1.62±0.59) times·min(-1)·mm(-1)] in the treatment group was significantly lower than the control group, P=0.034. After 72 h treatment, DEd [(41.4±8.1) vs (37.8±6.0) mm] was significantly higher than the control group, D-RSBI [(1.02±0.27) vs (1.22±0.43) times·min(-1)·mm(-1)] was significantly lower than the control group (all P<0.05). The average duration of NIV treatment time [(7.5±1.2) vs (9.3±2.6) h] in the treatment group was significantly shorter than that in the control group (P<0.01). There were no statistically significant differences in PaO(2), PCO(2), re-tracheal intubation rate and the mortality rate of 28 days. Conclusion: NIV combined with HFNCO sequential therapy can effectively relieve diaphragm fatigue and promote recovery of respiratory muscle strength, and it's better than NIV alone.