BACKGROUND:A large number of studies have confirmed that exosomes can promote osteogenesis and vascularization.However,simple exosome therapy has problems such as poor targeting,and the content of loaded molecules cannot reach the therapeutic concentration. OBJECTIVE:To load exosomes into injectable gluconolactone-sodium alginate β-tricalcium phosphate-polyethylene glycol hydrogel,and observe the effect of the hydrogel on peri-implant bone defect in vivo and in vitro. METHODS:Exosomes were extracted from bone marrow mesenchymal stem cells and wrapped in injectable gluconolactone-sodium alginate β-tricalcium phosphate-polyethylene glycol hydrogel.(1)In vitro experiment:The hydrogel loaded with exosomes and the hydrogel without exosomes were cocultured with endothelial progenitor cells,and exosomes uptake experiment,tubule formation experiment,cell proliferation,migration ability,and angiogenic gene detection were carried out.(2)In vivo experiment:Twelve male New Zealand white rabbits were used to prepare two standard implant cavities and corresponding bone defects in the long axis of one femur.A hydrogel loaded with exosomes was implanted in the bone defect after an implant was implanted in a cavity at the proximal end of the implant(experimental group),and an unloaded exosome hydrogel was implanted in the bone defect after an implant was implanted in a cavity at the distal end of the implant(control group).At 3,6 and 9 weeks after operation,bone defects with implants were removed and stained with hematoxylin-eosin staining and Masson staining.Simultaneously,osteogenic and angiogenic genes were detected at 9 weeks after operation. RESULTS AND CONCLUSION:(1)In vitro experiment:Exosomes could enter endothelial progenitor cells.The proliferation,migration,angiogenesis and gene(CD31,vascular endothelial growth factor and basic fibroblast growth factor)expression of endothelial progenitor cells in the hydrogel-loaded group were higher than those in the hydrogel-unloaded group(P<0.05).(2)In vivo experiment:Hematoxylin-eosin staining and Masson staining showed that at 3 weeks after operation,only a small amount of new bone was found in the two groups,and the material was partially degraded.At 6 weeks after operation,the amount of new bone in the two groups increased,and a large amount of new bone was found in the experimental group,with obvious calcium deposition.At 9 weeks after operation,compared with the control group,a large number of bone trabeculae thicker than mature were found in the experimental group,calcium salt deposition was more obvious,and a large number of osteoblasts were found around the bone trabeculae.The protein expressions of CD31,vascular endothelial growth factor,basic fibroblast growth factor,bone morphogenetic protein 2,type I collagen and osteocalcin in the experimental group were higher than those in the control group at 9 weeks after operation(P<0.05).(3)The exosome-loaded gluconolactone-sodium alginate β-tricalcium phosphate-polyethylene glycol hydrogel could promote the proliferation,migration and angiogenic differentiation of endothelial progenitor cells and promote the repair and regeneration of bone defects around implants.

ObjectiveTo investigate the influence of concentration ratio（CR） between the internal reference and target components on the quantitative accuracy of quantitative analysis of multi-components by single marker（QAMS） by taking ginsenosides as an example. MethodUltra performance liquid chromatography（UPLC） was employed， the contents of nine components in Ginseng Radix et Rhizoma（ginsenosides Rg₁， Re， Rf， Rh₁， Rb₁， Rc， Rb₂， Rb₃， Rd） were determined by external standard method（ES）. Using ginsenoside Rg₁ as the internal reference， the contents of the remaining 8 ginsenosides were determined by QAMS， and the quantitative results were compared with those of ES to evaluate the quantitative accuracy of the established QAMS. According to the contents of these 9 ginsenosides， the simulated sample solutions with different CRs of ginsenoside Rg₁ to ginsenosides Rf， Rb₂， Rd were prepared with the reference substance（CR=100∶1， 10∶1， 5∶1， 2∶1， 1∶1， 0.5∶1， 0.25∶1）， in order to validate the effect of the CRs between the internal reference and other components on the quantitative accuracy of the QAMS. ResultThe results of ginsenosides Re， Rf， Rb₁， Rc， Rb₂ calculated by the two methods were the same with the relative standard deviation（RSD）<3%， however， the results of ginsenosides Rh₁， Rb₃ and Rd calculated by the two methods were different with RSDs of 7.06%-9.61%. According to the result of the simulated sample solution， the difference between the calculated results of ES and QAMS was large when the CR between the internal reference（ginsenoside Rg₁） and other components was 5 or 10 or even higher. ConclusionThe quantitative error of QAMS will increase when the CR between the quantitative component and the internal reference is too large， so it is suggested that when establishing the QAMS， the components with close concentration to the internal reference should be selected for quantification.

Objective:To explore the effect of auricular acupressure therapy on postoperative pain and early rehabilitation of patients with total knee arthroplasty.Methods:From March to June 2021, 68 patients who underwent unilateral total knee arthroplasty for the first time under general anesthesia combined with peripheral nerve block in Liaocheng People's Hospital were selected by convenient sampling. According to the random number method, the patients were divided into observation group (35 cases) and control group (33 cases) . Both groups received routine pain management and early postoperative rehabilitation nursing after operation, and the Chinese medicine doctors pasted the Wangbuliuxing seed adhesive tape at the auricular points one day before operation. The control group did not take any pressing operation. The observation group was pressed at the time of 1 day before operation to 3 days after operation, 3 times a day. The pressing frequency was 60 times/min, and the pressing time was 1 minute per acupoint. The dosage of analgesics used immediately after leaving the Anesthesia Recovery Room and at 4, 24, 48 and 72 hours after operation, as well as the visual analog pain scores of rest and activity at each follow-up time point were recorded and compared between the two groups. The knee range of motion of the two groups on the second, the fourth and seventh days after operation was compared.Results:The dosage of patient-controlled intravenous analgesia analgesics in the observation group were lower than those in the control group at 24, 48 and 72 hours after operation, with statistically significant differences ( P<0.05) . The pain score of the patients in the observation group at rest 24 hours after operation was lower than that in the control group, and the pain scores of the patients in the active state at 24, 48 and 72 hours after operation were lower than those in the control group, and the differences were statistically significant ( P<0.05) . The knee range of motion in the observation group were higher than those in the control group on the second, fourth and seventh days after operation, and the differences were statistically significant ( P<0.05) . Conclusions:Auricular acupressure therapy can reduce the dosage of analgesics used after total knee arthroplasty, relieve the degree of pain, and promote the early recovery of patients.

Pharmaceutical analysis is a discipline based on chemical, physical, biological, and information technologies. At present, biotechnological analysis is a short branch in pharmaceutical analysis; however, bioanalysis is the basis and an important part of medicine. Biotechnological approaches can provide information on biological activity and even clinical efficacy and safety, which are important characteristics of drug quality. Because of their advantages in reflecting the overall biological effects or functions of drugs and providing visual and intuitive results, some biotechnological analysis methods have been gradually applied to pharmaceutical analysis from raw material to manufacturing and final product analysis, including DNA super-barcoding, DNA-based rapid detection, multiplex ligation-dependent probe amplification, hyperspectral imaging combined with artificial intelligence, 3D biologically printed organoids, omics-based artificial intelligence, microfluidic chips, organ-on-a-chip, signal transduction pathway-related reporter gene assays, and the zebrafish thrombosis model. The applications of these emerging biotechniques in pharmaceutical analysis have been discussed in this review.

Objective:To analyze the potential specificity of broad neutralizing antibodies (bNAbs) in one patient (DRVI01) with chronic HIV-1 infection.Methods:Sequences of the envelope glycoprotein (Env) obtained from DRVI01 at different time points were analyzed by comparing with the key amino acids of reported HIV-1 bNAbs in HIV Database. After reverse mutation of key amino acids that had frequently mutated to wild type, the neutralizing sensitivity of autologous plasma against wild-type and mutated Env-pesudoviruses was compared and the potential bNAbs in DRVI01 were speculated.Results:Reported key amino acids of 10 bNAbs classes were detected in 155 Env sequences derived from DRVI01. Frequent mutations were found in key amino acids of two bNAbs classes of gp41 fusion domain and gp120/gp41 interface. Neutralizing sensitivity of the contemporaneous autologous plasma and the plasma collected at the next time point against the mutated pesudoviruses was significantly increased as compared with wild-type pesudoviruses.Conclusions:Potential NAbs with similar key amino acids to those of gp41 fusion domain and gp120/gp41 interface might present in the HIV-1 infected patient with broad neutralizing antibodies.

Objective:To explore the influence of standardized and comprehensive incubational measures on perioperative treatment of extensively burned patients who underwent escharectomy and skin grafting.Methods:From January 2017 to November 2018, 50 patients with extensive burn who underwent escharectomy and skin grafting in the First Affiliated Hospital of Air Force Medical University and met the inclusion criteria of this study, were recruited in this retrospective cohort study. According to the incubational measures at that time, 20 patients (14 males and 6 females, aged (33.5±5.2) years) who received routine incubation during the perioperative period from January to October 2017 were set as routine incubation group, and 30 patients (23 males and 7 females, aged (35.8±1.4) years) who received standardized comprehensive incubational measures during the perioperative period from November 2017 to November 2018 were set as comprehensive incubation group. Their body temperature was controlled mainly in 4 stages: preoperative preparation and transfer from intensive care unit (ICU) to operating room, preoperative preparation in operating room, intraoperative operating room management, as well as postoperative transfer from operating room to ICU. The initial body temperature in operating room and intraoperative hypothermia duration, intraoperative blood loss, postoperative recovery time, postoperative chill, blister, and ulcer, and wound healing rate on post operation day (POD) 10 were recorded and calculated. Data were statistically analyzed with two independent samples t test and chi-square test. Results:(1) The initial body temperature in operating room of patients in comprehensive incubation group was (36.3±0.4) ℃, which was significantly higher than (35.6±0.4)℃ in routine incubation group, t=6.658, P<0.01; the intraoperative duration of hypothermia was (205±38) min, which was significantly shorter than (234±42) min in routine incubation group, t=2.564, P<0.05. (2) The intraoperative blood loss of patients in comprehensive incubation group was (323±114) mL, which was significantly less than (490±162) mL in routine incubation group, t=4.272, P<0.01; the postoperative recovery time was (36±8) min, which was significantly shorter than (49±17) min in routine incubation group, t=3.229, P<0.01. (3) The incidence of postoperative chill of patients in comprehensive incubation group was significantly lower than that in routine incubation group ( χ2=28.626, P<0.01). The incidences of postoperative blister and ulcer of patients between the 2 groups were close. (4) On POD 10, the wound healing rate of patients in comprehensive incubation group was (78.08±0.06)%, which was significantly higher than (71.03±0.08)% in routine incubation group, t=3.694, P<0.01. Conclusions:The standardized and comprehensive incubational measures can effectively improve the initial body temperature of patients entering the operating room, shorten the intraoperative duration of hypothermia, reduce the amount of blood loss and postoperative complications, as well as shorten the postoperative recovery time, thus improve the wound healing rate.

Objective To study the effect of autologous periosteum on the healing of tendon-bone interface in rabbit rotator cuff tear. Methods Supraspinatus tenotomy was performed on one side shoulder in 60 New Zealand white rabbits to establish the model of rotator cuff tear. The rabbits were randomly divided into two groups:the study group(used autologous periosteum to promote the suture fixation)and control group(simple suture fixation). The rabbits were sacrificed at 4,8 and 12 weeks postoperatively with twenty rabbits sacrificed each time. Tissue samples of the tendon-bone interface was taken for histological examination and biomechanical test was performed to assess the strength of tendon-bone junction. Results At 4 weeks the study group showed extensive inflammatory cell infiltration, and a small amount of chondrocytes and extracellular matrix. At 8 weeks, the study group showed a large amount of immature chondrocytes arranged rather regularly. At 12 weeks,the tendon bone junction appeared similar to normal. In the study group,both the tendon-bone connection and the arrangement of chondrocytes were significantly better than the control group. The result of biomechanical testing showed that the highest tendon load in the study group was significantly higher than the control group (P < 0.05). Conclusions The use of autologous periosteum as a patch to strengthen the repair of the rotator cuff tear can effectively promote the healing of tendon bone interface, shorten the rotator cuff healing time and has good biological properties. This method provides an experimental basis for clinical rotator cuff repair surgery.

Objective:To optimize the water extraction technology of Xiaokeantangfang. Methods: The extraction technology of Xiaokeantangfang was optimized by orthogonal design with the content of astragaloside A and the extract yield as the indices and the a-mount of added water,extraction time and extraction times as the influencing factors,and the results were verified. Results:The amount of added water had significant effect on the content of astragaloside A(P<0. 05). The optimal extraction technology was as follows:12-fold amount of added water,extracting for 3 times with 0. 5 hour per time. The average extract yield was 38. 33%(RSD=1. 22%,n=3), and the average content of astragaloside A was 0. 0828 mg·g-1 for 3 batches of samples in the validation tests(RSD=3. 37%, n=3). Conclusion:The optimal water extraction technology of Xiaokeantangfang is stable and feasible, which can provide experimen-tal basis for the oral preparation of Xiaokeantangfang.

Objective This study was performed to investigate the effect of hypoxia on glucose-6-phosphate isomerase (G6PI) expression and cell cycle of fibroblast-like synoviocytes from synovium of rheumatoid arthritis (RA) and osteoarthritis (OA) under hypoxia or normoxia.Methods Fibroblast-like synoviocytes were cultured with either of hypoxia (3％ oxygen) or normoxia (21％ oxygen) for 24 hours.The mRNA expression of G6PI and HIF-1α was tested by PCR quantification,while the protein levels of G6PI and HIF-1α were measured by western blot.Cell cycle was performed by FACS.T-test and Mann-Whitney U were used for statistical analysis.Results The expression levels of G6PI mRNA under hypoxia in RA were higher than those of OA (2.6±0.4 vs 1.5±0.4,P＜0.05).The protein levels of G6PI in RA were higher than those of OA (P＜0.05).The expression levels of HIF-1α mRNA under hypoxia in RA were higher than those of OA (2.9±0.8vs 1.4 ±0.4,P＜0.05).The protein levels of HIF-1α in RA were higher than those of OA (P＜0.05).The G1 phase ratio of cell cycle was decreased significantly under hypoxia than those of normoxia in RA ELs (t=1 1.31,P＜0.05).The S and G2 phase ratio of cell cycle were increased.Conclusion Hypoxia upregulates G6PI and HIF-1α expression and improves proliferation in fibroblast-like synoviocytes.

Background and purpose: Because of the aggressive nature of hilar cholangiocarcinoma and the absence of effective adjuvant therapy, surgical radical resection offers hilar cholangiocarcinoma patients the only choice. Research focus include preoperative assessment, the use of preoperative biliary drainage, the range of hepatic resection, and the range of lymphadenectomy. To investigate the clinical experience and efifcacy of combined hepatectomy in the treatment of hilar cholangiocarcinoma. Methods: Two hundred and seven patients with hilar cholangiocarcinoma treated surgically in the First Afifliated Hospital of Kunming Medical University form Jan. 2007 to Oct. 2013 were retrospectively analyzed. Results:Of the 207 patients, 125 patients who received radical resection (R0 resection) and the curative resection rate was 60.4%. One hundred and iffty-six cases were treated in combined hepatectomy group, 51 cases in non-hepatectomy group, the rate of R0 resection was 70.5%in hepatectomy group and 29.4%in non-hepatectomy group, and the difference was signiifcant (P<0.01). Two patients died perioperatively, the main postoperative complications included hepatic function insufifciency and bile leakage. One hundred and seventy-two patients were followed up, the median survival time of the 102 patients who received R0 resection was 45 months, and the 1, 3, 5 year survival rates were 96.1%, 59.1%and 17.2%. The median survival time of the 70 patients who received R1-2 resection was 26 months, and the 1, 3 year survival rates were 81.3%and 19.2%, and none of the patient survived for over 5 years. The survival rate of patients who received R0 resection was signiifcantly higher than those who received R1-2 resection (χ2=39.121, P<0.01). In the hepatectomy group was awarded the R0 resection in patients with postoperative 1, 3, 5 year survival rate was 97.8%, 63.9% and 18.0%, in non-hepatectomy group received R0 resection in patients with postoperative 1, 3, 5 year survival rate was 83.3%, 20.8%and 8.3%. There were signiifcant differences in the postoperative survival rate between both group (χ2=5.988, P=0.014). Conclusion:Radical excision is the key to improve the long term survival. Combined hemihepatectomy and standardized lymph node resection has signiifcantly improved the radical resection rate and the efifcacy of treatment for hilar cholangiocarcinoma.