Sjögren's syndrome (SS) is an autoimmune disease characterized primarily by oral and periocular dryness. Astragalus-Salvia (AS) and Ophiopogon-Dendrobium (OD) represent two frequently utilized herb pairs in SS treatment. While the combination of AS-OD herb pairs demonstrates clinical efficacy in alleviating SS symptoms, its underlying mechanism remains unclear. This investigation sought to assess the therapeutic effects and elucidate the potential mechanisms of AS-OD in non-obese diabetic (NOD)/Ltj mice with SS. The study utilized NOD/Ltj mice as SS models, administering AS-OD treatment for 10 weeks at doses of 113.1, 226.2, and 339.3 mg·d^-1·20 g^-1. Results demonstrated that AS-OD improved SS symptoms, evidenced by enhanced salivary flow rate, decreased anti-SSA/Ro and anti-SSB/La antibody levels, increased swimming duration, and reduced lactate (LA) and blood urea nitrogen (BUN) levels in NOD/Ltj mice. AS-OD reduced lymphocyte infiltration, enhanced Aquaporin-5 (AQP5) expression in the submandibular gland, decreased inflammatory cytokine levels in the submandibular gland, and reduced the T helper type 17/regulatory T lymphocyte (Th17/Treg) cell ratio in the spleen. Transcriptomic and proteomic analyses indicated AS-OD's involvement in regulating phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) and Janus kinase 3/signal transducer and activator of transcription 3 (JAK1/STAT3) pathways, with inhibitory effects validated in both NOD/Ltj mice submandibular gland and A-253 cells. Furthermore, AS-OD enhanced cell viability and reduced A-253 cell apoptosis through the PI3K/AKT pathway. In A-253 cells, AS-OD reduced inflammatory cytokine levels, CXC chemokine ligand 9/10 (CXCL9/10), and T-cell chemotaxis by inhibiting the JAK1/STAT3 pathway. AS-OD mitigates SS by suppressing inflammation and immune responses through the PI3K/AKT and JAK1/STAT3 pathways.
Animals ; STAT3 Transcription Factor/genetics* ; Sjogren's Syndrome/immunology* ; Mice, Inbred NOD ; Proto-Oncogene Proteins c-akt/genetics* ; Phosphatidylinositol 3-Kinases/genetics* ; Mice ; Drugs, Chinese Herbal/administration & dosage* ; Signal Transduction/drug effects* ; Janus Kinase 1/genetics* ; Humans ; Female ; Astragalus Plant/chemistry* ; Male

As main recipient cells for porcine reproductive and respiratory syndrome virus (PRRSV), porcine alveolar macrophage (PAM) are involved in the progress of several highly pathogenic virus infections. However, due to the fact that the PAM cells can only be obtained from primary tissues, research on PAM-based virus-host interactions remains challenging. The improvement of induced pluripotent stem cells (iPSCs) technology provides a new strategy to develop IPSCs-derived PAM cells. Since the CD163 is a macrophage-specific marker and a validated receptor essential for PRRSV infection, generation of stable porcine induced pluripotent stem cells lines containing CD163 reporter system play important roles in the investigation of IPSCs-PAM transition and PAM-based virus-host interaction. Based on the CRISPR/Cas9- mediated gene editing system, we designed a sgRNA targeting CD163 locus and constructed the corresponding donor vectors. To test whether this reporter system has the expected function, the reporter system was introduced into primary PAM cells to detect the expression of RFP. To validate the low effect on stem cell pluripotency, we generated porcine iPSC lines containing CD163 reporter and assessed the pluripotency through multiple assays such as alkaline phosphatase staining, immunofluorescent staining, and EdU staining. The red-fluorescent protein (RFP) expression was detected in CD163-edited PAM cells, suggesting that our reporter system indeed has the ability to reflect the expression of gene CD163. Compared with wild-type (WT) iPSCs, the CD163 reporter-iPSCs display similar pluripotency-associated transcription factors expression. Besides, cells with the reporter system showed consistent cell morphology and proliferation ability as compared to WT iPSCs, indicating that the edited-cells have no effect on stem cell pluripotency. In conclusion, we generated porcine iPSCs that contain a CD163 reporter system. Our results demonstrated that this reporter system was functional and safe. This study provides a platform to investigate the iPS-PAM development and virus-host interaction in PAM cells.
Swine ; Animals ; Induced Pluripotent Stem Cells/metabolism* ; Receptors, Cell Surface/genetics* ; Antigens, CD/metabolism* ; Porcine respiratory and reproductive syndrome virus/genetics*

Objective:To investigate the therapeutic effect and possible mechanism of low intensity pulsed ultrasound（LIPUS）on painful gait in mice with Achilles tendon injury.Methods:Male C57BL/6 mice were selected to establish the Achilles tendon injury model by surgically full-thinkness tear of the right Achilles tendon. The mice were divided into Achilles tendon injury group and Achilles tendon injury+LIPUS group according to the random number table method，with 7 mice per group. The Catwalk gait analysis system was used to evaluate the gait function of the mice by measuring the following five parameters 14 days after operation，including print area，standing time，step cycle，max intensity and stride length. Morphological changes of the Achilles tendon were observed by HE staining. Immunofluorescence staining was used to detect the expression level of nitric oxide synthase（iNOS）in the Achilles tendon tissues. At the same time，Achilles tendon cells were isolated and cultured in vitro. The cells were induced by 100 ng/ml lipopolysaccharide（LPS）for 12 hours to establish in vitro model（LPS group），and treated with LIPUS（LPS+LIPUS group）. The control group was set as well（without any treatment）. The nuclear translocation of nuclear transcription factor-κB（NF-κB）P65 was detected by immunofluorescence staining，and the expression of iNOS and phosphorylation（p）-NF-κB p65 protein was detected by Western blotting. Results:Compared with Achilles tendon injury group［（0.14±0.10）cm 2］，the print area of the affected limb in Achilles tendon injury+LIPUS group［（0.28±0.13）cm 2］was increased（ P<0.05）. Compared with Achilles tendon injury group［（0.11±0.04）seconds］，the standing time of the affected limb in Achilles tendon injury+LIPUS group［（0.21±0.03）seconds］was increased（ P<0.05）. Compared with Achilles tendon injury group［（0.25±0.05）seconds］，the step cycle of the affected limb in Achilles tendon injury+LIPUS group［（0.40±0.05）seconds］was increased（ P<0.05）. There was no significant difference in the max intensity and stride length between Achilles tendon injury group and Achilles tendon injury+LIPUS group（ P>0.05）. HE staining showed obvious hyperplasia of Achilles tendon tissues in Achilles tendon injury group，with loosely and randomly arranged fibers，accompanied by neovascularization and inflammatory cell infiltration. Achilles tendon injury+LIPUS group showed more orderly arranged fibers in Achilles tendon tissues，and the degree of neovascularization and inflammatory cell infiltration were reduced. Compared with Achilles tendon injury group［（5.70±0.81）%］，the expression level of iNOS in Achilles tendon injury+LIPUS group［（2.84±0.94）%］was decreased 14 days after operation（ P<0.05）. Immunofluorescence results of Achilles tendon cells showed that NF-κB p65 protein entered the nucleus in LPS group compared with control group，and that LIPUS treatment inhibited nuclear translocation. In control group，Western blotting showed that iNOS was not expressed，and that the expression of phosphorylated NF-κB p65 was 0.63±0.16. Compared with control group，the expression levels of iNOS（0.99±0.22）and p-NF-κB P65（1.02±0.19）in LPS group were significantly increased（ P<0.05）. Compared with LPS group，the expression levels of iNOS（0.62±0.10）and p-NF-κB P65（0.65±0.21）in LPS+LIPUS group were decreased（ P<0.05）. Conclusion:LIPUS treatment can alleviate pain gait in mice with Achilles tendon injury and inhibit iNOS expression in vivo and in vitro，which may be related to inhibition of NF-κB signaling pathway.

Impaired awareness of hypoglycemia is an acquired complication of insulin therapy, which affects people with type 1 and insulin-treated type 2 diabetes mellitus, whereby the ability to perceive the onset of hypoglycemia becomes diminished or absent, which not only affects life health, but also causes fear of hypoglycemia , and causing a serious burden on the quality of life. The paper reviews the concept,the assessment tools,the occurrence,the influencing factors and intervention strategies of impaired awareness of hypoglycemia.In order to provide suggestions for the researches of impaired awareness of hypoglycemia in China.

Objective To explore the effect of the intervention of clinical pharmacists on the rational use of piperacillin-tazobactam by using PDCA cycle, in order to provide reference for rational drug use. Methods The problems of piperacillin-tazobactam in our hospital was analyzed. PDCA cycle was used to manage the problems. Then, the data before and after PDCA cycle was compared and analyzed. Results After using PDCA cycle, the irrational use rate of piperacillin-tazobactam gradually decreased, from 9% in February 2018 to 2% in February 2019; the doses decreased from 4380 in February 2018 to 3346 in February 2019; and the frequency of usage decreased from 391 DDDs in February 2018 to 298 DDDs in February 2019. The effectiveness and continuous improvement of PDCA cycle in managing piperacillin-tazobactam were significant. Conclusion PDCA cycle can effectively improve the management effectiveness of piperacillin-tazobactam administration.

Objective? To construct the critical illness patient inter-hospital referral process of primary hospitals based on Delphi method so as to provide a basis for the inter-hospital critical illness patient standardized referral process in the medical alliance model. Methods? The inter-hospital referral process was designed by retrieving literatures at home and abroad, and was demonstrated and revised with the modified Delphi method. Results? We constructed a total of 30 items from four aspects including work preparation of medical staff as well as reception medical staff before inter-hospital referral, risk-prevention during referral and patients' condition after referral. Among two rounds of expert consultation, the active coefficients were all 100% and the authority coefficients were all 0.843 respectively. Conclusions? The critical illness patient inter-hospital referral process of primary hospitals based on modified Delphi method accords with the principle of scientific nature and operability which has positive a meaning for standardizing clinical inter-hospital critical illness patient referral.

Objective To explore the effects of referral and rehabilitation status of patients with total knee arthroplasty (TKA) under the mode of medical alliance.Methods One hundred and ninety cases with total knee arthroplasty in the secondary rehabilitation hospitals after referral from a tertiary hospital from January 2016 to January 2017 were analyzed, including general demographic data, disease and surgical data, rehabilitation data after referral, hospital day and change of medical expenses of the tertiary hospital and rehabilitation hospitals before and after the promotion of the medical alliance mode.Results After receiving the rehabilitation treatment in secondary rehabilitation hospitals, the range of motion, muscle strength and some other indices of the patients' surgical legs were significantly improved (P<0.05). And the hospital day of the surgical hospital was shorten from (15.05±6.59) d to (12.97±5.24) d, and medical expenses were decreased by 2.18% in total, 4.76% per knee and 14.01% in rehabilitation costs.Conclusions Under the mode of medical alliance, transferring patients with TKA from a tertiary hospital to secondary rehabilitation hospitals in their early postoperative phase can facilitate recovery of the patients, and it can also help to reduce hospital day and medical expenses, thus to optimize the allocation of medical resources.

Objective To introduce a practical method that can be used to efficiently express,purify and identify Alzheimer's disease (AD) related beta-site app-cleaving enzyme 1 (BACE1) in common eukaryotic cells.Methods BACE1 cDNA was fished out from human brain cDNA library and ligated into the pEGFP-c3 expression vector,and then,the recombinant plasmid was transfected into the HEK293 cells.The BACE1 protein was purified with TALON Mental Affinity Resins column.The target protein was identified by Western blotting and fluorescence resonance energy transfer (FRET).BACE1 Activity Assay Kit was employed to test the activity of purified BACE1 in vitro.The recombinant BACE1/pEGFP-c3 plasmid and amyloid precusor protein (APP)/pDsRed-Monomer-N1 plasmid were co-transfected to the HEK293 cells and the cleavage activity of BACE1 in the cells was identified by Western blotting.Results The sequencing data of the obtained BACE1 gene were identical with those in GenBank.Activity test showed that the fluorescent values of blank controls,expressed BACE1 and standard BACE1 were 55.013±3.597,1836.629±154.195 (n=3) and 2639.548±207.1901 (n=3),respectively;as compared with the control group,significant differences were noted in both of the two groups (F=78.681,P=0.000);however,there is no significant difference between expressed BACE1 and standard BACE1 groups (P＞0.05).Westem blotting showed the co-transfected BACE1 could cleave APP in HEK293 cells and the CTF-APP band was detectable.Conclusion A practical protocol is established for high expression,purification and identification of BACE1 in HEK293 cells,which is helpful to obtain BACE1,an important molecular target in AD research and treatment.

Aim To investigate the effect of simvastatin ( Sim ) on endogenous antioxidant system after acute myocardial infarction ( AMI ) and its potential mecha-nisms. Methods The acute myocardial infarction ( AMI ) rat models were made by ligation left anterior descending of coronary artery. Then the successful models were randomly divided into myocardial infarc-tion group ( MI group) and simvastatin group ( Sim,20 mg·kg-1·d-1), another group without ligation left anterior descending of coronary artery served as sham group(Sham group). The Sim group was administered simvastatin by gavage for 7 days. MI group and Sham group received saline. Hemodynamic parameters, lipid levels, troponinI ( c-TnI ) and lactate dehydrogenase ( LDH) concentrations were examined after 7days, and the levels of superoxide dismutase ( SOD) and glutathi-one peroxidase ( GP) of myocardial antioxidant system were detected by ELISA. The expression of cardiac p-Akt and p-eNOS protein were detected by Western blot. Results Acute myocardial infarction significant-ly lowered cardiac hemodynamic parameters, increased serum c-TnI and LDH levels, lowered levels of SOD and GP, and lowered the expression of p-Akt and p-eNOS protein. However, Sim could effectively prevent the deterioration of cardiac function, reduce serum c-TnI and LDH levels, increase levels of SOD and GP, and increase p-Akt and p-eNOS protein expression. Conclusion Early using Sim can effectively improve heart function after acute myocardial infarction, acti-vate myocardial antioxidant system,and reduce myocar-dial necrosis, which may be related to increasing the expression of p-Akt and p-eNOS.

Compared to the mature monitoring and reporting system of ADRs in developed countries such as the USA and EU,medical institutions in China have played too much a role.Instead,local primary monitoring and reporting system of ADRs should expand their scope of monitoring and reporting,as drug makers,chain drugstores and retailers should develop an effective ADR monitor/report system as well.At present,such a primary system is plagued by insufficient budget and staff,imbalanced urban/rural layout and defective statements.Recommended measures include enhanced surveillance for drug makers,chain drugstores and retailers,for higher reporting ratio of ADRs at primary level; greater publicity and better monitoring and penalty mechanism; better platform for various organizations.