italic>α-Conotoxin ArIB[V11L,V16D] is currently the most optimal selective inhibitor of α7 nicotinic acetylcholine receptor (nAChR) known. In order to explore chemical modification methods and enrich its application in targeting nAChR, this study utilized the linker to covalently connect camptothecin and 7-amino-4-methylcoumarin to the [2,4] disulfide bond of ArIB[V11L,V16D]. Therefore, two peptide-drug conjugates (PDCs), ArIB[V11L,V16D]-5 and ArIB[V11L,V16D]-6, and one fluorescent-labeled peptide, ArIB[V11L,V16D]-7 were constructed. Cytotoxicity evaluation showed that the IC₅₀ values against non-small cell lung cancer cell line A549 of the two PDCs were respectively 1.3 and 4.1 times of camptothecin, indicating slight reduction in activity at the cellular level which was related to the linker structure. Fluorescence spectrum scanning revealed that the excitation and emission wavelength of the fluorescent-labeled peptide were 340 nm and 403 nm respectively, and the fluorescence features of 7-amino-4-methylcoumarin as a marker were retained without fluorescence quenching. This modification strategy laid a solid foundation for the further application of α-conotoxin ArIB[V11L,V16D] in PDCs and fluorescent probes.

Objective To analyze the expression profile of circular RNA (circRNA) in LPS-treated murine peritoneal macrophages (MPMs) and to investigate the effects of mmu_circ_0000790 (circ790) on the secretion of pro-inflammatory cytokines in MPMs following LPS stimulation.Methods MPMs were isolated from C57BL/6 male mice and then stimulated with (LPS treatment group) or without (blank control group) 1 mg/L of LPS for 12 hours.Supernatants of cell culture and cell pellets were collected from each group.Enzyme linked immunosorbent assay (ELISA) was used to measure the changes in the secretion of IL-1β,IL-6 and TNF-α in the supernatants.Expression profile of circRNA in macrophages from the two groups (n=3) was analyze by circRNA microarray.Some differentially expressed circRNAs were validated by real-time PCR (RT-PCR).Moreover,RT-PCR was also performed to detect the expression of circ790 in MPMs after LPS stimulation.Small interfering RNA (siRNA) oligo specific for circ790 was designed and synthesized.Then the synthesized siRNA oligo and normal control (NC) oligo were transfected into MPMs by LipofectamineTM2000.The transfected MPMs were treated with LPS.ELISA was used to detect the levels of IL-1β,IL-6 and TNF-α in the supernatants.Arraystar's home-made microRNA target prediction software was used to predict circ790/microRNA.Results The concentrations of IL-1β,IL-6 and TNF-α in the LPS treatment group were significantly higher than those in the blank control group (P<0.01).These results indicated that the inflammatory model of MPMs was successfully constructed.Statistical differences in 34 differentially expressed circRNAs were found between the two groups (P<0.05).Among them,12 circRNAs were up-regulated and the other 22 circRNAs were down-regulated in the LPS treatment group.RT-PCR results were generally consistent with the microarray data.The expression of circ790 was increased by (1.94±0.15),(3.18±0.13) and (4.21±0.22) folds as compared with that of the blank control group at 3,6 and 12 h after LPS stimulation (P<0.05).After transfecting the MPMs with circ790-siRNA,the secretion of IL-6 was significantly decreased (P<0.05) without notable influence on the secretion of IL-1β and TNF-α.circ790 could function as a microRNA sponge to regulate the gene expression.Conclusion These data show a significantly altered circRNA expression profile in the LPS-treated MPMs.circ790 may be involved in the regulation of IL-6 secreted by macrophages.

Nicotinic acetylcholine receptors (nAChRs) belong to ligand-gated ion channel receptors, of which α7 nAChR subtype is widely distributed in the cerebral cortex, thalamus, hippocampus, and also identified in microglia, macrophages, bone marrow cells, etc. Previous studies revealed that α7 nAChR is closely related to the function of the cholinergic anti-inflammatory pathway, and is a vital target for drug development of Alzheimer's disease and schizophrenia. The establishment of a stable α7 nAChR in vitro drug screening system is crucial for the efficient screening of novel drugs targeting this target. Recombinant expression of different subtypes of nAChRs on Xenopus laevis oocyte membranes and current detected by two-electrode voltage clamp (TEVC) is an advanced and complex model for novel drug screening. Molecular chaperones can assist the assembly of some nAChR subunits to form functional receptors, providing a stable expression model for the screening of compounds targeting this receptor. In this study, a molecular chaperone gene of α7 nAChR, transmembrane protein 35A (Tmem35a), was isolated and cloned from rats. We constructed the recombinant expression vector and obtained the cRNA of Tmem35a by in vitro transcription technique. Two cRNAs (Tmem35a and α7) were mixed and injected into X. laevis oocytes for expression. Then, the effects of this molecular chaperone on the current expression and pharmacological properties of α7 nAChR were evaluated by the TEVC. The results revealed that TMEM35A, also known as novel acetylcholine receptor chaperone (NACHO) could effectively increase the expression of α7 nAChR protein on oocyte membranes, and the amount of α7 nAChR protein was increased about 1-fold. The peak current induced by agonist acetylcholine (ACh) was increased about 10-fold. After injection of Tmem35a cRNA, the median effect concentration (EC₅₀) value of α7 nAChR to agonist ACh is 228.5 μmol·L^-1, which shows almost no difference from native α7 nAChR (EC₅₀: 223.3 μmol·L^-1), indicating the preservation of the normal properties of α7 nAChR. The results of this investigation indicate that the molecular chaperone NACHO effectively assists the heterologous expression of α7 nAChR in X. laevis oocytes, which provides a model for screening the potency of lead compounds targeting α7 nAChR. All animal experiments in this study were reviewed and approved by the Ethics Committee of Guangxi University (approval number: GXU-2023-0249).

N-type voltage-gated calcium (Ca²⁺) channels (N-type VGCC, Ca_V2.2) mediate Ca²⁺ influx in response to action potential at the presynaptic terminal, and play an important role in synaptogenesis, neurotransmitter release and nociceptive signal transduction. It is a new target for the development of drugs for the treatment of neuralgia (chronic pain) and other major diseases. Due to the difficulty of calcium channel expression in vitro and the detection of channel current, there is a great lack of new drug screening models. In this study, we established and optimized the electrophysiological drug screening model using Xenopus laevis oocytes for the recombinant expression of Ca_V2.2 in vitro (this study were reviewed and approved by the Ethics Committee of Guangxi University, approval number: GXU-2023-0249). Firstly, the linear plasmids encoding cDNA of major subunit α1B and auxiliary subunits α2δ1 and β3 of rat Ca_V2.2 were used as templates for in vitro transcription to generate their related mRNA (cRNA), after which three kinds of cRNA were injected into Xenopus laevis oocytes at the mass ratio of 2∶1∶1 for expression. The two-electrode voltage clamp (TEVC) technique was used to detect the inward current produced by Ca_V2.2. At the same time, the expression conditions of Ca_V2.2 were optimized, and its gating function was characterized from the aspects of channel activation and inactivation. The results showed that 3-5 days after cRNA microinjection, stable Ca_V2.2-mediated barium ion (Ba²⁺) currents were successfully detected. The interference of endogenous potassium channels and Ca²⁺-activated chloride channels can be eliminated by tetraethylammonium hydroxide (TEAOH) and 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis (BAPTA-AM) treatment. The maximum potential for Ca_V2.2 activation is 0 mV, and the current reverses to be outward when the membrane potential is greater than +50 mV. By fitting the steady-state activation and inactivation curves, the half-maximal activation potential and half-maximal inactivation potential of Ca_V2.2 are identified as -15.9 and -60.2 mV. In this study, a stable Ca_V2.2 expression system was established based on Xenopus laevis oocytes. The in vitro expression system can provide a new way for the screening of Ca_V2.2 active compounds or lead drugs.

Objective To assess the feasibility of semi-quantitative scoring system for contrast-enhanced ultrasound (CEUS)quantitative analysis's color images in the differential diagnosis of breast nodules.Methods Totally 244 BI-RADS 4 breast solid lesions received CEUS before core needle biopsy or surgical resection were included.A semi-quantitative scoring system for color images of CEUS quantitative analysis were built.The scores were given as follows:1 )Color type and its distribution (0 to 4);2)Color scope (0 to 1 );3)Color margin (0 to 1 );4)Color shape (0 to1 ).The total score for each lesion would be from 0 to 7.And the differenital value between benign and malignant lesions were assessed.Results The total semi-quantitative scores of 102 malignant tumors (5.1 ±1 .7)was significant higher than that of benign lesions (3.34±0.7)(P < 0.05 ).In 102 malignant lesions,the total scores of 81 lesions (79.41 %)were more than 4 points,and in 142 benign lesions,the total scores of 89 lesions (62.67%)were less than 4 points.Depending on the Wilcox rank sum test (Mann-Whitney)analysis,the distribution of total scores between benign and malignant lesions was significant different (P <0.000 1).Total score 4 was selected as the best cutoff,the area under ROC curve was 0.749,on which the sensitivity,specificity and accuracy were 79.4%,62.7% and 69.67%,respectively.Conclusions The semi-quantitative scoring system of CEUS quantitative analysis color images showed good sensitivity but not satisfied specificity and accuracy in differential diagnosis between malignant and benign breast lesions.

The cyanuric chloride linkers have been used for cyclizing polypeptide, but not used for α-conotoxin, the peptides with rich disulfide bonds and more amino acid residues. In this study, cyclic peptides c[A10L]PnIA-1-4 were synthesized efficiently by lysine assisted cyanuric chloride linkers with 28.92%-52.00% yields. The activity evaluation showed that the IC₅₀ values of c[A10L]PnIA-1 against α7 and α3β2 nAChR subtypes were 5 and 7 times higher than [A10L]PnIA respectively, and the subtype selectivity was maintained. The results of circular dichroism show that this cyclization method had no significant effect on its secondary structure. Compared with the commonly used head-to-tail cyclization in conotoxin cyclization, this method has the advantages of rapid reaction and high yield, which is expected to be further applied to the cyclization study of various α-conotoxins.

BACKGROUNDOn May 12, 2008, a major earthquake hit Wenchuan County in Sichuan Province of China. The number of cases of crush injury following this event was high. Ultrasonic appearance of rhabdomyolysis (RM) caused by crush injury in the Wenchuan earthquake was observed to evaluate the diagnostic value of ultrasound for detection of rhabdomyolysis.

METHODSWe analyzed clinical and ultrasonic manifestations of 50 cases of RM and 18 cases of RM with osteofascial compartment syndrome (OCS). All cases were caused by crush injury in the Wenchuan earthquake. For these RM patients, we also evaluated the correlations between creatine kinase (CK) and the scope of the muscle lesions as observed by ultrasound.

RESULTSThere were differences in clinical symptoms, physical signs and ultrasonic appearance between the two groups of patients. The ultrasonic characteristics of the RM were as follows: the striated muscle in the lesions thickened with good overall continuity, and the muscle texture was vague; the strength of the echo was uneven and the echo was cloudy or ground glass-like. Liquid dark zones appeared between muscles and were spindle-like or irregular in shape. There were no blood flow signals in the liquid dark areas. The volume of the striated muscle increased in patients with OCS; the fascia wrapping the muscle showed arched protrusions and significant displacement. The flow velocity of the distal arteries decreased and the spectrum was abnormal. The muscle lesion scope of RM group and RM and OCS group was (7.8 +/- 2.0) cm and (13.6 +/- 3.1) cm, respectively. The correlation coefficient (r) between the muscle lesion scope and the CK was 0.681 for the RM group (P < 0.05) and 0.516 for the RM and OCS group (P < 0.05).

CONCLUSIONSThe ultrasonogram of RM has characteristic manifestations and can provide important information for clinical diagnosis and treatment of rhabdomyolysis.

Adolescent ; Adult ; China ; Compartment Syndromes ; diagnostic imaging ; Crush Syndrome ; diagnostic imaging ; Earthquakes ; Female ; Humans ; Male ; Rhabdomyolysis ; diagnostic imaging ; Ultrasonography ; Young Adult

Objective:To evaluate the effect of small-dose furosemide on tissue edema after shoulder arthroscopy.Methods:A total of 368 patients of either sex, aged 18-65 yr, of American Society of Anesthesiology physical status Ⅰ orⅡ, were divided into 2 groups ( n=184 each) by a random number table method: control group (group C) and small-dose furosemide group (group F). A posterior approach to interscalene brachial plexus block was selected, tracheal intubation was performed under general anesthesia, and all the patients were placed in the lateral position.At 20-30 min before the end of surgery, patients in group F received intravenous injection of furosemide 2 mg (diluted to 2 ml in normal saline) and patients in group C received intravenous injection of normal saline 2 ml.The tissue thickness from the surface of the second rib of the middle clavicular line to the skin (CR2) was measured by ultrasound immediately after nerve block (T 0), immediately after tracheal intubation (T 1), immediately after the end of operation (T 2), and at 30 min and 1, 4 and 8 h after operation (T 3-6). Arterial blood gas analysis was performed at T 1-3, Hct and blood glucose concentration were measured simultaneously, and relative blood volume was calculated.Tube malposition at T 2 and hypoxemia within 30 min after operation were recorded. Results:Compared with the baseline at T 0, CR2 on the affected side was significantly increased at T 2-5 in group F, and CR2 on the affected side was significantly increased at T 2-6 in group C ( P<0.05). CR2 at T 2-5 was significantly higher on the affected side than on the healthy side ( P<0.05). Compared with group C, intraoperative urine volume was significantly increased, CR2 was decreased at T 2-5, the incidence of tube malposition and hypoxemia was decreased ( P<0.05), and no significant changes were found in pH value, PaO 2, PaCO 2, Na + , K + , Ca 2+ , Cl -, blood glucose concentration or relative blood volume at each time point in group F ( P>0.05). Conclusion:Small-dose furosemide can reduce tissue edema after shoulder arthroscopy and decrease the occurrence of postoperative tube malposition and hypoxemia, without causing disturbances in the internal environment.

Antibiotic-associated diarrhea(AAD)in children is a type of diarrhea that occurs after the use of antibiotics in children,and its pathogenesis is closely related to the intestinal flora.The medication of antibiotics can affect the metabolic function of the intestinal flora and the immune function of the body,and then leads to the occurrence of AAD.In the view of Chinese medicine,AAD in children is mainly involved the spleen,and the etiology of the disease is due to the weakness of the spleen and stomach of the body constitution together with the attack of the pestilential pathogen and the accumulation of drug toxin.The pathogenesis of ADD in children is characterized by spleen deficiency with predominant dampness,deficiency of spleen qi,and insufficiency of spleen yang.Spleen deficiency is the root cause of pediatric AAD,and spleen and intestinal flora have commonality,so the treatment of pediatric AAD can be performed from the perspective of the spleen.The treatment of pediatric ADD from the spleen follows the principle of strengthening and activating the spleen,and the regulation of the spleen for achieving the purpose of treating the disease from the root can be achieved by the methods of strengthening spleen and draining dampness,strengthening spleen and replenishing qi,and strengthening spleen and warming yang separately with the fundamental prescriptions of Shenlin Baizhu Powder,Sijunzi Decoction,and Fuzi Lizhong Pills.

OBJECTIVETo study the clinicopathologic features of various types of mature T-cell and natural killer (NK)/T-cell lymphoma in Guangdong, China, with respect to the 2008 WHO classification of lymphoid neoplasms.

METHODSEleven hundred and thirty-seven (1137) cases of mature T-cell or NK/T-cell lymphoma diagnosed during the period from 2002 to 2006 in Guangzhou area were retrieved. The clinical data, histologic features and immunohistochemical findings were reviewed by a panel of experienced hematopathologists. Additional immunostaining was performed if indicated. The cases were re-classified according to the 2008 WHO classification of lymphoid neoplasms.

RESULTSNine hundred and sixty-three (963) cases fulfilled the diagnostic criteria of mature T-cell or NK/T-cell lymphoma and accounted for 20.1% of all cases of lymphoma encountered during the same period (963/4801). A predominance of extranodal involvement was noted in 644 cases (66.9%), while 319 cases (33.1%) showed mainly nodal disease. The prevalence of various lymphoma subtypes was as follows: peripheral T-cell lymphoma, unspecified (PTCL, NOS) 293 cases (30.4%), extranodal NK/T-cell lymphoma, nasal type 281 cases (29.2%), anaplastic large cell lymphoma (ALCL) 198 cases (20.6%), and angioimmunoblastic T-cell lymphoma (AILT) 46 cases (4.8%). The male-to-female ratio was 1.99. The median age of the patients was 44 years, with the peak age of PTCL, NOS, extranodal NK/T-cell lymphoma, nasal type and AILT being 55 to 64 years, 25 to 54 years and 65 to 74 years, respectively. ALK-positive ALCL occurred more frequently in young age, while the ALK-negative ALCL cases occurred mainly in the elderly.

CONCLUSIONSExtranodal lesions predominate in mature T-cell and NK/T-cell lymphomas occurring in Guangzhou area. There is a male predominance and the overall incidence shows no increasing trend with age of the patient. The peak age of various subtypes however varies. The most common subtype was PTCL, NOS, followed by extranodal NK/T-cell lymphoma, nasal type, ALCL and AILT. The relatively frequent occurrence of extranodal NK/T-cell lymphoma, nasal type in Guangdong area is likely associated with the high incidence of Epstein-Barr virus infection there.

Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; Epstein-Barr Virus Infections ; Female ; Humans ; Immunoblastic Lymphadenopathy ; metabolism ; pathology ; virology ; Infant ; Lymphoma, Extranodal NK-T-Cell ; metabolism ; pathology ; virology ; Lymphoma, Large-Cell, Anaplastic ; metabolism ; pathology ; virology ; Lymphoma, T-Cell ; classification ; metabolism ; pathology ; virology ; Lymphoma, T-Cell, Peripheral ; metabolism ; pathology ; virology ; Male ; Middle Aged ; Protein-Tyrosine Kinases ; metabolism ; Receptor Protein-Tyrosine Kinases ; Retrospective Studies ; Sex Factors ; World Health Organization ; Young Adult