convalescents, and to screen for broad-spectrum neutralizing antibodies against the SARS-CoV-2 RBD. Methods Using biotinylated RBD as a molecular probe, flow cytometry was employed to perform single-cell sorting of B cells from peripheral blood mononuclear cells (PBMCs) of convalescents. The obtained B cells were lysed and subjected to reverse transcription, followed by nested PCR amplification of the heavy and light chains of antibodies was conducted using random primers. The amplified products were cloned into corresponding expression vectors, and the respective matched heavy-light chain plasmids were co-transfected into 293F cells for expression. Monoclonal antibodies were then purified using Protein A column chromatography. Neutralization experiments were conducted with the wild-type (WT) pseudovirus, and antibodies with IC50<0.1 μg/mL were selected for further testing of neutralizing breadth and potency against the wild-type (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and currently prevalent pseudovirus strains (XBB, BA.5, BF.7). Results A total of 21 RBD-specific monoclonal B cells were obtained from two recovered patients, resulting in the isolation of 13 pairs of antibody light/heavy chains. Nine antibodies were successfully expressed, with P1-A1, P1-B6, and P1-B9 exhibiting IC50 values below 0.1 μg/mL against the pseudovirus of the wild-type strain (WT). Specifically, P1-B6 effectively neutralized the wild-type strain (WT), Beta variant (B.1.351), and Delta variant (B.1.617.2), with IC50 values reaching 0.01 μg/mL. P1-B9 demonstrated effective neutralization against the wild-type strain (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and Gamma variant (P.1) pseudoviruses, with IC50 values of 0.42 μg/mL, 0.63 μg/mL, 0.28 μg/mL, and 2.50 μg/mL, respectively. Additionally, P1-B6 exhibited good neutralization against BA.5 and BF.7 pseudoviruses, with IC50 values of 0.06 μg/mL and 0.09 μg/mL, respectively. Conclusions Infection with the SARS-CoV-2 WT strain can induce the generation of neutralizing antibodies with broad-spectrum activity. Generating these broadly neutralizing antibodies does not require an excessively high somatic hypermutation. The obtained antibodies can be used as candidates for SARS-CoV-2 diagnosis and prevention.

Premature ovarian failure （POF） is a common gynecological endocrine disease ，which can cause follicular development stagnation or premature failure in the ovary ，leading to ovarian function loss ，and seriously affecting women ’s physical and mental health . Traditional Chinese medicine shows less toxicity and side effects but significant effect in the treatment of POF . Chinese herb monomers are the active ingredients of traditional Chinese medicine that exert their effects ，and their chemical structure are determined . From the perspective of Chinese herb monomers ，the author reviewed the mechanism of their effect on improving POF . The results showed that icariin ，quercetin，resveratrol and ginsenoside Rg 1 could improve the ovarian function of POF model animals by activating the phosphatidylinositol 3-kinase/protein kinase B signaling pathway ，inhibiting oxidative stress and apoptosis of granulosa cells ，promoting ovarian autophagy ，and regulating the level of sex hormones ；puerarin and resveratrol can improve the ovarian function of POF model by interfering with Wnt/ β-catenin signaling pathway ，reducing oxidative stress response ，and inhibiting follicular atresia . Although the Chinese herb monomers have certain effect in improving POF，most of them remain in the experimental stage ，and the clinical efficacy in improving POF cannot be objectively evaluated . In the future ，the development of new dosage forms of Chinese herb monomers should be strengthened to lay a foundation for the development of new drugs for treating POF .

Objective To explore the mechanism of latent human immunodeficiency ciency virus type 1 (HIV-1) infection is unclear, especially in dendritic cells (DC).We hypothesized that DC-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) binds with HIV-1 may activate HIV-1 provirus.Methods We generated a model by transfecting 293T cells with a DC-SIGN expression plasmid and a HIV-1 5'long terminal repeat (LTR) reporter plasmid, and then stimulated the 293T cells with HIV-1 gp120 protein, wild-type HIV-1 and VSV-G-pNL4.3 pseudotype virus ( without gp120 protein).CEM-Bru cells were transfected with the DC-SIGN expression plasmid and stimulated by HIV-1 gp120 protein.Then HIV-1 replication was detected.The involvement of the ERK, p38 and NF-κB pathways signaling in this response were determined by inhibiting the pathways specifically and detecting the phosphorylation of the signaling kinase.Results The HIV-1 5'LTR was reactivated by HIV-1 gp120 in DC-SIGN-expressing 293T cells.After HIV-1 gp120 protein stimulation of the mold of CEM-Bru cells, the increasing expression of HIV-1 Tat mRNA and HIV-1 p24,which implies early and late HIV-1 provirus replication was reactivated by the HIV-1 gp120/DC-SIGN stimulation.HIV-1 gp120/DC-SIGN stimulation reactivates latent HIV-1 provirus via the NF-κB signal pathway.Conclusion HIV-1 gp120/DC-SIGN stimulation reactivates latent HIV-1 provirus via the NF-κB signal pathway.

Objective To observe the inhibitory effects of sub-MIC matrine alone and in combination with erythromycin on Staphylococcus epidermidis biofilms and their influences on morphological changes of the biofilms.Methods Minimum inhibitory concentrations (MIC) of matrine and erythromycin against Staphylococcus epidermidis were determined by the serial dilution method,antibacterial activity of matrine combined with erythromycin against planktonic S.epidermidis was evaluated by the checkerboard method.S.epidermidis biofilms were constructed in vitro,XTT reduction assay was used to evaluate influences of sub-MIC matrine alone and in combination with erythromycin on metabolism and adhesion of S.epidermidis biofilms,and scanning electronic microscope(SEM) was applied to observe the morphological and the structural changes of the biofilms.Results The MIC of erythromycin to S.epidermidis was 7.8125 μg/ml,while the MIC of matrine was greater than 1000 μg/ml,besides,a synergistic effect between erythronmycin and matrine on planktonic S.epidermidis was shown (FIC＜0.5).The sub-MIC matrine had no significant inhibitory effect on adhesion of S.epidermidis,and also the combination of the two agents was better than was used alone.However,the sub-MIC matrine had inhibitory effects on metabolism and morphology of S.epidermidis biofilms,and the combination of the two agents was weaker than was used alone.Conclusion Both the sub-MIC matrine and erythromycin had a significant inhibitory effect on S.epidermidis biofilm formation.Combination of the two agents showed synergistic effects on plankton and adhesion of S.epidermidis,but showed no synergistic effect on metabolism and morphology of the biofilms.

Objective To investigate the impact on ovarian reserve function by different hemostasis methods during laparoscopic surgery in treatment of ovarian endometrioma.Methods From September 2008 to February 2010,162 cases with ovarian endometrioma undergoing laparoscopic surgery in Shandong Provincial Hospital were enrolled in this study.At the 3rd day of the menstrual cycle before surgery and the 1 st,3rd,6th and 12th cycle after surgery,serum FSH and anti-mullerian hormone(AMH) and ultrasound basal antral follicle count (AFC) and peak systolic velocity (PSV) were examined and compared.Based on hemostasis method,those patients were divided into 3 groups,including 54 cases in bipolar hemostasis,54 cases in ultrasonic scalpel hemostasis and suture after excision of endometrioma.Results (1) Before surgery:no significant different factors among three groups before surgery were observed,including age,size of endometrioma,the level of FSH,AMH,AFC,PSV (P ＞ 0.05).(2) Ovarian reserve function after surgery:①FSH:at the 1st,3rd,6th and 12th month follow-up,the FSH in the bipolar group was (11.7 ±4.0),(9.9 ± 4.0),(9.5 ± 4.3),(9.5 ± 3.9) U/L,and the FSH in ultrasonic scalpel group was (11.4 ±4.3),(9.7 ± 4.0),(9.2 ± 3.7),(9.9 ± 4.6) U/L,were significantly higher than (9.3 ± 3.8),(6.7 ±3.0),(6.5 ± 3.2),(6.4 ± 2.2) U/L in suture group respectively (all P ＜ 0.05).()AMH:at the 1 st,3rd,6th and 12th month follow-up,the AMH in the bipolar group was (1.8 ±0.9),(1.8 ± 1.0),(1.9 ±1.0),(2.0 ± 1.0) μg/L,and the AMH in the ultrasonic scalpel group was (1.6 ±± 0.8),(1.8 ± 1.0),(2.0 ± 1.1),(2.1 ± 1.0) μg/L,which were significantly lower than (2.8 ± 1.7),(2.9 ± 1.6),(3.0 ±1.3),(3.2 ± 1.5) μg/L in suture group,respectively (all P ＜ 0.05).③AFC:there was no significant difference of APC among the three groups in the 1st month after surgery.However,at the 3rd,6th and 12th month follow-up,the AFC of 4.8 ± 1.4,5.9 ± 1.5,6.1 ± 1.5 in the suture group was significant higher than 3.7 ± 1.4,4.1 ± 1.4,4.0 ± 1.5 in bipolar group and 3.6 ± 1.3,4.0 ± 1.1,3.9 ± 1.5 in ultrasonic group,respectively (all P ＜ 0.05).④PSV:at the 1 st,3rd,6th and 12th month follow-up,the PSV of the bipolar group(7.9 ±3.5),(8.1 ±3.3),(8.4 ±3.1),(8.6±3.0) cm/s in bipolar group and (8.1 ±3.5),(8.0 ± 3.0),(7.9 ± 3.2),(8.0 ± 2.9) cm/s in ultrasonic group were significant lower than (10.9 ± 3.3),(12.0 ± 3.2),(11.8 ± 3.0),(12.1 ± 4.1) cm/s in suture group,respectively.(allP＜0.05).Conclusions Bipolar or ultrasonic scalpel hemostasis during laparoscopic excision of ovarian endometrioma is associated with a significant reduction in ovarian reserve.Electrocoagulation of the ovarian tissue should be avoided.

Objective To detect the role of AP-1 or ETS-1 transcription factor binding sites (TFBS) in activity of DC-specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN) pro-moter. Methods Genomic DNA was extracted from peripheral blood. Complete DC-SIGN promoter and those without AP-1 or ETS-1 TFBS were amplified by PCR using designed primers. The PCR products were digested with MLu Ⅰ and Bgl Ⅱ and then ligated into MLu Ⅰ and Bgl Ⅱ digested pGL-3/Basic and pGL-3/En-hancer vectors. Transfection in Hacat and 293 cells was accomplished with Trans Fast liposome. Activity of luciferase was detected after 48 h. Results The DC-SIGN promoters without AP-1 or ETS-1 TFBS and the recombined pGL-3 vectors were correctly constructed. The activity of DC-SIGN promoters without AP-1 was reduced 20% (293) and 10% (Hacat), which was 40%-50% with enhancer. The activity of DC-SIGN pro-moters without ETS-1 was nearly vanished, no matter with or without enhancer. Conclusion ETS-1 TFBS, not AP-1 TFBS, plays an important role in activity of DC-SIGN promoter.

Self-assembly of peptides is ubiquitous in the body of creatures. The molecules of peptides combine with each other to form proteins with different functions through self-assembly. The formation of a specific conformation of one type of protein is owing to the self-assembly of its compositive amino acids. So, researchers can design self-assembly of peptides at the molecular level and can control its formation and configuration. It has the potential for application in the preparation of new medicines and biomaterials. In recent years, self-assembling peptides have been increasingly high-lighted and used to simulate the function of natural biomolecules, to synthesize peptide-medicine, and to serve as the carriers of medicine.
Biocompatible Materials ; chemical synthesis ; Molecular Conformation ; Nanotechnology ; methods ; Peptides ; chemistry ; Protein Engineering ; methods

OBJECTIVETo determine the relationships of Met416Val and XbaI polymorphism of muscle glycogen synthase (GYS1) gene and Trg64Arg variant of the beta(3)-adrenergic-receptor (beta(3)-AR) gene with type 2 diabetes mellitus (DM) and its intermediate phenotypes in the Chinese population.

METHODSPolymerase chain reaction-oligonucleotide ligation assay and restriction fragment length polymorphism assay were used to evaluate the GYS1 and beta(3)-AR gene polymorphisms in 102 pairs of case-control Chinese spouses.

RESULTSSubjects with Met416Val variant had a significantly higher 2-hour post-glucose level than subjects without this variant had in diabetic group (P = 0.032). The Met416Val polymorphism of GYS1 gene was not significantly associated with the risk of type 2 DM (adjusted OR = 1.67; 95% CI: 0.73 - 3.81, P = 0.223). Subjects with Trp64Arg variant had a significantly higher serum uric acid level than subjects without this variant had in diabetic group (P = 0.034). The combination of BMI and Arg64 allele carrier of the beta(3)-AR gene increased the diabetic risk over four-fold (adjusted OR = 4.00; 95% CI: 1.53 - 10.45, P = 0.005).

CONCLUSIONSIn the Chinese population, Met416Val polymorphism is identified in a subgroup of diabetic subjects with high 2-hour post-glucose. It will explain why some diabetic patients appear to be genetically predisposed to developing high postpradial glucose level. The presence of the Arg64 allele in the beta(3)-AR gene may predispose patients to higher serum uric acid level.

Adult ; Aged ; Alleles ; Body Mass Index ; Diabetes Mellitus, Type 2 ; blood ; genetics ; Glycogen Synthase ; genetics ; Humans ; Hyperglycemia ; genetics ; Middle Aged ; Polymorphism, Genetic ; Postprandial Period ; physiology ; Receptors, Adrenergic, beta-3 ; genetics ; Uric Acid ; blood

Objective To detect the relationship between the polymorphism of the glycogen-targeting regulatory subunit of the skeletal muscle glycogen-associated protein phosphatase 1 (PPP1R3) gene and type 2 diabetes by case-control study. Methods We genotyped the PPP1R3 gene Asp905Tyr polymorphism and a common 3'-untranslated region AT (AU)-rich element (ARE) polymorphism in 101 type 2 diabetic patients and 101controls by oligonucleotide ligation assay (OLA) and polyacrylamide gel elecrophoresis, respectively. Results Subjects with Tyr/Tyr genotypes whose body mass index (BMI)<25 were used as the reference group. Those whose BMI25 with Asp905 had a 3.66-fold increase (95% CI: 1.48-9.06, P=0.005) in type 2 diabetes risk. No association was found between 3'UTR ARE polymorphism and type 2 diabetes mellitus (OR=1.15; 95% CI: 0.62-2.14, P=0.65). Conclusion A joint effect between the Asp905 and BMI increases the risk of type 2 diabetes, and Asp905Tyr and ARE polymorphism of PPP1R3 gene are not the major diabetogenic gene variants in Chinese population.

Objective To investigate the effect of high dose mifepristone and high dose progesterone in the treatment of patients with endometrial carcinoma and to explore the possible mechanisms associating with them Methods Thirty untreated patients diagnosed as endometrial carcinoma through dilation and curettage of the uteri were divided into 3 groups at random Each group was given medroxyprogesterone acetate(MPA),(500 mg/day) or mifepristone(MIF),(100 mg/day)or MIF(100 mg/day)+ MPA(500 mg/day)for 5 days respectively On the sixth day, hysterectomy was performed on these patients The endometrial cancer specimen of post hysterectomy was compared with the one of pre administrating The morphologic changes of the endometrial cancer cells were observed through light microscope Immunohistochemistry assay (SP method) was applied to determine the localization and immunoreactive intensity of proliferating cell nuclear antigen(PCNA), estrogen receptor (ER), progesterone receptor (PR), B cell leukemia lymphoma 2 (bcl 2), bcl 2 associated X protein(bax) and CD 44 v6 Results Better differentiation degree and active excretion were observed in all of the post hysterectomy endometrial specimen In the same time, apoptosis of carcinoma cells was observed The most significant changes were seen in the MIF+MPA group In the MPA group,the pre treatment and post treatment expression of PR(2 9?1 1,1 6?0 8),ER(2 8?0 9,1 4?0 9),PCNA(0 84?0 10,0 60?0 12),bcl 2(0 236?0 089,0 157?0 981) and CD 44 v6 (4 6?1 8,2 5?1 9) were all decreased(all P 0 05) In the MIF+MPA group, the expression of PR(3 2?1 0,0 8?0 8),ER(2 7?0 9,0 7 ?0 9 ),PCNA(0 81?0 09,0 25?0 09),bcl 2(0 225?0 091,0 066?0 009)and CD 44 v6(4 5?1 9,2 7?1 6) were all decreased(all P