Single hormone or synthetic agonist induces Gs/Gi coupling selectivity of EP receptors via distinct binding modes and propagating paths.

To accomplish concerted physiological reactions, nature has diversified functions of a single hormone at at least two primary levels: 1) Different receptors recognize the same hormone, and 2) different cellular effectors couple to the same hormone-receptor pair [R.P. Xiao, Sci STKE 2001, re15 (2001); L. Hein, J. D. Altman, B.K. Kobilka, Nature 402, 181-184 (1999); Y. Daaka, L. M. Luttrell, R. J. Lefkowitz, Nature 390, 88-91 (1997)]. Not only these questions lie in the heart of hormone actions and receptor signaling but also dissecting mechanisms underlying these questions could offer therapeutic routes for refractory diseases, such as kidney injury (KI) or X-linked nephrogenic diabetes insipidus (NDI). Here, we identified that Gs-biased signaling, but not Gi activation downstream of EP4, showed beneficial effects for both KI and NDI treatments. Notably, by solving Cryo-electron microscope (cryo-EM) structures of EP3-Gi, EP4-Gs, and EP4-Gi in complex with endogenous prostaglandin E2 (PGE2)or two synthetic agonists and comparing with PGE2-EP2-Gs structures, we found that unique primary sequences of prostaglandin E2 receptor (EP) receptors and distinct conformational states of the EP4 ligand pocket govern the Gs/Gi transducer coupling selectivity through different structural propagation paths, especially via TM6 and TM7, to generate selective cytoplasmic structural features. In particular, the orientation of the PGE2 ω-chain and two distinct pockets encompassing agonist L902688 of EP4 were differentiated by their Gs/Gi coupling ability. Further, we identified common and distinct features of cytoplasmic side of EP receptors for Gs/Gi coupling and provide a structural basis for selective and biased agonist design of EP4 with therapeutic potential.

Effect of acupuncture at Weizhong (BL 40) on bladder urination function based on ultrasound / 中国针灸

OBJECTIVE@#To observe the effect of acupuncture at Weizhong (BL 40) with deqi on bladder urination function.@*METHODS@#A total of 60 healthy subjects were randomized into an observation group and a control group, 30 subjects in each group. Under the guidance of ultrasound, acupuncture was applied Weizhong (BL 40) on both sides. In the observation group, the needling depth was reached to the tibial nerve, and lifting-thrusting twirling method was used to induce deqi. In the control group, the needling depth was reached to the superficial fascia, and no manipulation was operated to induce deqi. The needles were retained for 10 min and acupuncture was given once in both groups. The bilateral ureteral ejection frequency and volume of the bladder were observed by ultrasound before and after acupuncture, and the score of clinical evaluation scale of deqi sensation was observed in both groups.@*RESULTS@#After acupuncture, the frequency of bilateral ureteral ejection in the observation group and the bladder volume in the two groups were increased compared before acupuncture (P<0.05), and the frequency of bilateral ureteral ejection, bladder volume and score of clinical evaluation scale of deqi sensation in the observation group were higher than those in the control group (P<0.05, P<0.01).@*CONCLUSION@#Acupuncture at Weizhong (BL 40) with deqi improves the bladder urination function. Ultrasound visualization improves the standardization and safety of acupuncture, intuitively evaluates the acupuncture effect, and provides an objective basis for the correlation between meridian points specificity and zang-fu organs.

Endogenous Lipid-GPR120 Signaling Modulates Pancreatic Islet Homeostasis to Different Extents.

Long-chain fatty acids (LCFAs) are not only energy sources but also serve as signaling molecules. GPR120, an LCFA receptor, plays key roles in maintaining metabolic homeostasis. However, whether endogenous ligand-GPR120 circuits exist and how such circuits function in pancreatic islets are unclear. Here, we found that endogenous GPR120 activity in pancreatic δ-cells modulated islet functions. At least two unsaturated LCFAs, oleic acid (OA) and linoleic acid (LA), were identified as GPR120 agonists within pancreatic islets. These two LCFAs promoted insulin secretion by inhibiting somatostatin secretion and showed bias activation of GPR120 in a model system. Compared with OA, LA exerted higher potency in promoting insulin secretion, which is dependent on ß-arrestin2 function. Moreover, GPR120 signaling was impaired in the diabetic db/db model, and replenishing OA and LA improved islet function in both the db/db and streptozotocin-treated diabetic models. Consistently, the administration of LA improved glucose metabolism in db/db mice. Collectively, our results reveal that endogenous LCFA-GPR120 circuits exist and modulate homeostasis in pancreatic islets. The contributions of phenotype differences caused by different LCFA-GPR120 circuits within islets highlight the roles of fine-tuned ligand-receptor signaling networks in maintaining islet homeostasis.

Research on mechanism of salidroside activating HIF-1 a signaling pathway to promote MOB proliferation / 中国药理学通报

Aim To explore the effeet of salidroside (SAL) on proliferation of osteoblasts and its possible mechanism by using mouse primary osteoblasts ( MOB) as the research object.Methods Alkaline phosphatase ( ALP) staining was used to identify the extracted primary cells.MTT was used to detect the effect of SAL on the proliferation.RT-PCR, Western blot, ELISA were used to investigate the molecular mechanism of SAL.Results The extracted cells generated black-brown deposits by ALP staining, which were shown to be osteoblasts clearly.SAL promoted the pro liferation of MOB.Meanwhile, SAL could up-regulate the mRNA and protein expression levels of hypoxia-inducible factor-lcx ( HIF-1 ex), vascular endothelial growth factor ( VEGF ) , angiopoietin-like protein 4 (ANGPTL4) and interleukin 6 ( IL-6 ) , which were down-regulated by using the HIF-lex blocker YC-1.Conclusions SAL could promote the proliferation of MOB through HIF-1 a/VEGF, ANGPTL4, IL-6 signaling pathway.

Preventing and treating anterior commissure adhesion with mucosal flap: a study in canines and clinical cases / 中华耳鼻咽喉头颈外科杂志

Objective: To evaluate the effectiveness of mucosal flap combined with silicone keel for preventing and treating anterior commissure adhesion in canines and clinical cases. Methods: A prospective experiment was performed from November 2019 to June 2021. Twenty five canines were randomly divided into 5 groups(A, B, C, D, E). Group A, B, C, D received anterior commissure injury by CO2 laser, then separately treated with free mucosal flap-keel complex,intralaryngeal mucosal flap-keel complex, silicone keels and without treatment, group E didn't injure the vocal cord after intubation. The keel was removed after 2 weeks, the larynx was harvested after 4 weeks. The effectiveness of anterior commissure adhesion prevention was evaluated by manifestation under laryngoscope, standard vocal cord length and standard glottic area. A retrospective analysis was performed on sixteen patients with anterior commissure lesion, who underwent mucosal flap-keel technique in Huashan Hospital of Fudan University from January 2019 to January 2021 (10 cases with free mucosal flap-keel complex and 6 cases with intralaryngeal mucosal flap-keel complex). All the patients underwent evaluation of laryngeal function included manifestation under laryngoscope each month and voice analysis before and 3 month after surgery. SPSS 20.0 software was used for statistical analysis. Results: No surgery accident or complication happened in canines and patients. The standard vocal cord length and standard glottic area after 4 weeks in group B were significantly higher than those in group A, C, D (Hstandard vocal cord length=31.688, Hstandard glottic area=16.444, P<0.05). The standard vocal cord length and standard glottic area after 4 weeks in group A were also significantly higher than those in group C, D(Hstandard vocal cord length=20.936, Hstandard glottic area=11.786, P<0.05). The standard vocal cord length and standard glottic area after 4 weeks in group A, B, E were not significantly different to that before surgery(tA left standard vocal cord length=2.636, tA right standard vocal cord length=2.582, tB left standard vocal cord length=2.707, tB right standard vocal cord length=2.673, tE left standard vocal cord length=0.370, tE right standard vocal cord length=0.821, tA standard glottic area=2.731, tB standard glottic area=2.753, tE standard glottic area=-0.529, P>0.05). The standard vocal cord length and standard glottic area after 4 weeks in group C, D were significantly lower than those before surgery(tC left standard vocal cord length=16.137, tC right standard vocal cord length=13.984, tD left standard vocal cord length=11.903, tD right standard vocal cord length=14.587, tC standard glottic area=10.280, tD standard glottic area=22.974, P<0.05). During 6-18 months of follow-up in clinical patients, no one developed a glottic web. Three months after surgery, Jitter, Shimmer, noise to harmonic ratio(NHR), the maximum phonation time(MPT)in all patients were significantly different from preoperative(tintralaryngeal mucosal flap jitter=24.885, tintralaryngeal mucosal flap shimmer=22.643, tintralaryngeal mucosal flap NHR=6.202, tintralaryngeal mucosal flap MPT=-9.661, tfree mucosal flap jitter=25.459, tfree mucosal flap shimmer=18.683, tfree mucosal flap NHR=5.705, tfree mucosal flap MPT=-20.840, P<0.05). Conclusion: Mucosal flap combined with silicone keel is an effective technique for preventing and treating anterior commissure adhesion. The effect of pedicled intralaryngea lmucosal flap is better.

Planning of ecologically suitable areas for Ligusticum chuanxiong under background of soil cadmium pollution / 中国中药杂志

Chuanxiong Rhizoma is a traditional Chinese medicinal material mainly produced in Sichuan and Chongqing of China. In recent years, the cadmium content in Chuanxiong Rhizoma produced in most of the genuine producing areas has exceeded the standard, which makes Chuanxiong Rhizoma difficult to be exported. To solve the problem of excessive cadmium content in soil, this study employed the MaxEnt model to simulate the potential geographic distribution of Ligusticum chuanxiong and evaluate important environmental factors, and re-plan its ecologically suitable areas based on the mineral distribution characteristics and soil cadmium pollution status. The results showed that the places suitable for L. chuanxiong growing covered an area of 335 523.69 km~2, mainly in central and eastern Sichuan, southern Shaanxi and most parts of Chongqing. Among them, the highly suitable areas of L. chuanxiong were mainly concentrated in Chengdu, Ya'an, Deyang, and Mianyang. Solar radiation, annual precipitation, and annual range of temperature were evaluated as important variables affecting the distribution of L. chuanxiong, with the contribution rates of 62.3%, 13.3%, and 6.8%, respectively. In addition, Qionglai county, Chongqing county, Mianyang city(Youxian district and Fucheng district), Qingchuan county, and Xinjin county were classified into the first-class ecologically suitable zone, covering a total area of 2 768.87 km~2. The se-cond-class ecologically suitable zone was even wider, involving such counties as Tongjiang county, Renshou county, Jianyang county, and Nanjiang county, and the total area reached 43 616.92 km~2. The re-planning of the ecologically suitable areas for L. chuanxiong has provided strong data support for the cultivation and resource development of L. chuanxiong and also new ideas for solving the problem of excessive cadmium content in L. chuanxiong.

Genomic evidence for divergent co-infections of SARS-CoV-2 lineages

Recently, patients co-infected by two SARS-CoV-2 lineages have been sporadically reported. Concerns are raised because previous studies have demonstrated co-infection may contribute to the recombination of RNA viruses and cause severe clinic symptoms. In this study, we have estimated the compositional lineage(s), tendentiousness, and frequency of co-infection events in population from a large-scale genomic analysis for SARS-CoV-2 patients. SARS-CoV-2 lineage(s) infected in each sample have been recognized from the assignment of within-host site variations into lineage-defined feature variations by introducing a hypergeometric distribution method. Of all the 29,993 samples, 53 (~0.18%) co-infection events have been identified. Apart from 52 co-infections with two SARS-CoV-2 lineages, one sample with co-infections of three SARS-CoV-2 lineages was firstly identified. As expected, the co-infection events mainly happened in the regions where have co-existed more than two dominant SARS-CoV-2 lineages. However, co-infection of two sub-lineages in Delta lineage were detected as well. Our results provide a useful reference framework for the high throughput detecting of SARS-CoV-2 co-infection events in the Next Generation Sequencing (NGS) data. Although low in average rate, the co-infection events showed an increasing tendency with the increased diversity of SARS-CoV-2. And considering the large base of SARS-CoV-2 infections globally, co-infected patients would be a nonnegligible population. Thus, more clinical research is urgently needed on these patients.

Structural studies of phosphorylation-dependent interactions between the V2R receptor and arrestin-2.

Arrestins recognize different receptor phosphorylation patterns and convert this information to selective arrestin functions to expand the functional diversity of the G protein-coupled receptor (GPCR) superfamilies. However, the principles governing arrestin-phospho-receptor interactions, as well as the contribution of each single phospho-interaction to selective arrestin structural and functional states, are undefined. Here, we determined the crystal structures of arrestin2 in complex with four different phosphopeptides derived from the vasopressin receptor-2 (V2R) C-tail. A comparison of these four crystal structures with previously solved Arrestin2 structures demonstrated that a single phospho-interaction change results in measurable conformational changes at remote sites in the complex. This conformational bias introduced by specific phosphorylation patterns was further inspected by FRET and 1H NMR spectrum analysis facilitated via genetic code expansion. Moreover, an interdependent phospho-binding mechanism of phospho-receptor-arrestin interactions between different phospho-interaction sites was unexpectedly revealed. Taken together, our results provide evidence showing that phospho-interaction changes at different arrestin sites can elicit changes in affinity and structural states at remote sites, which correlate with selective arrestin functions.

Ganoderic acid hinders renal fibrosis via suppressing the TGF-ß/Smad and MAPK signaling pathways.

Renal fibrosis is considered as the pathway of almost all kinds of chronic kidney diseases (CKD) to the end stage of renal diseases (ESRD). Ganoderic acid (GA) is a group of lanostane triterpenes isolated from Ganoderma lucidum, which has shown a variety of pharmacological activities. In this study we investigated whether GA exerted antirenal fibrosis effect in a unilateral ureteral obstruction (UUO) mouse model. After UUO surgery, the mice were treated with GA (3.125, 12.5, and 50 mg· kg-1 ·d-1, ip) for 7 or 14 days. Then the mice were sacrificed for collecting blood and kidneys. We showed that GA treatment dose-dependently attenuated UUO-induced tubular injury and renal fibrosis; GA (50 mg· kg-1 ·d-1) significantly ameliorated renal disfunction during fibrosis progression. We further revealed that GA treatment inhibited the extracellular matrix (ECM) deposition in the kidney by suppressing the expression of fibronectin, mainly through hindering the over activation of TGF-ß/Smad signaling. On the other hand, GA treatment significantly decreased the expression of mesenchymal cell markers alpha-smooth muscle actin (α-SMA) and vimentin, and upregulated E-cadherin expression in the kidney, suggesting the suppression of tubular epithelial-mesenchymal transition (EMT) partially via inhibiting both TGF-ß/Smad and MAPK (ERK, JNK, p38) signaling pathways. The inhibitory effects of GA on TGF-ß/Smad and MAPK signaling pathways were confirmed in TGF-ß1-stimulated HK-2 cell model. GA-A, a GA monomer, was identified as a potent inhibitor on renal fibrosis in vitro. These data demonstrate that GA or GA-A might be developed as a potential therapeutic agent in the treatment of renal fibrosis.

Epidemiological analysis of novel coronavirus pneumonia in Huangpu District of Shanghai / 上海预防医学

Objective To understand the epidemiological characteristics of COVID-19 epidemic in Huangpu District of Shanghai, and to provide scientific evidence for prevention and control of COVID-19. Methods Descriptive statistics were used to study the suspected and confirmed cases of COVID-19 reported from January 21 through March 10, 2020 in Huangpu District, Shanghai. Results A total of 120 suspected cases of COVID-19 were reported, of which 12 were diagnosed and 108 were excluded.The first confirmed case was reported on January 21, and the last case was on February 10; the majority (11/12) of the confirmed cases were reported from January 21 through February 1.The average duration of time from the symptom onset to the first medical visit was 2.6 days, whereas the average duration from the first medical visit to the hospital diagnosis was 2.2 days.There were 15 suspected cases with a confirmed history of residence or tourism in Wuhan, in which 6 were confirmed cases.Moreover, 5 suspected cases had a confirmed history of contact with other confirmed cases, in which 3 were confirmed cases.Thus, exposure in Wuhan and exposure to confirmed cases were the most significant risk factors at this stage of the epidemic. Conclusion The 12 cases identified in Huangpu District of Shanghai are all adults, half of whom had confirmed history of exposure in Wuhan.The first cluster of COVID-19 cases in Shanghai is documented in Huangpu District.Epidemiological investigation reveals that the confirmed cases might be infectious the day before the symptom onset.

Genetically Encoded Fluorescent Amino Acid for Monitoring Protein Interactions through FRET.

Förster resonance energy transfer (FRET) is a well-established method for studying macromolecular interactions and conformational changes within proteins. Such a method normally uses fluorescent proteins or chemical-labeling methods which are often only accessible to surface-exposed residues and risk-disturbing target protein structures. Here, we demonstrate that the genetic incorporation of a synthetic fluorescent amino acid, L-(7-hydroxycoumarin-4-yl) ethylglycine (Cou) and natural endogenous fluorophore Tryptophan (Trp) residues of a protein could serve as an efficient FRET pair to monitor protein interactions, using the signaling transducer ß-arrestin-1 as a model system. We used this technology to record the dynamic spectra in both binding and competition experiments of ß-arrestin-1, the contribution of each specific phosphate in ternary complex formation, in a rapid and efficient manner. The determined Kd value for the association between the active arrestin and Fab30 is 0.68 µM in the three-component interaction system. Moreover, we were able to determine the contributions of the site 3 phospho-site and the site 6 phospho-site binding, each contributing to the high affinity ternary complex assembly as 2.7 fold and 15.5 fold, respectively, which were never determined before. These results thus highlighted the potential usage of this new method in measurement of the allosteric-induced enhanced affinity with small amount proteins and in a fast manner and in a complex system. Collectively, our newly developed Trp:Cou FRET system based on genetic expansion technology has extended the molecular toolboxes available for biochemical and structural biology studies.

Research Progress of Angiogenesis in Atherosclerotic Plaque in Chinese Medicine and Western Medicine / 中国结合医学杂志

Angiogenesis in atherosclerotic plaque plays a critical role in the mechanism of atherosclerotic physiopathology. Present consensus shows that angiogenesis in atherosclerotic plaque is mainly resulted in hypoxia, inflammation and some pro-angiogenic factors. The homeostasis in plaque, which is hypoxic and infiltrated by inflammatory cells, may lead to angiogenesis, increase the plaque instability and the incidence rate of vascular events. This article reviews the progression of pathogenetic mechanism, physiopathological significance, relevant detecting technique and corresponding therapeutic methods of Chinese and Western medicine of angiogenesis in atherosclerotic plaque, so as to provide more theoretical basis for atherosclerotic clinical treatment.