Objective To develop and verify a reversed phase high-performance liquid chromatography method for the determination of the purity of recombinant Mycobacterium tuberculosis(Mtb)Ag85b protein stock solution.Methods Fourfactor,three-level orthogonal test was designed,with the area,trailing factor,peak area and peak area RSD as the evaluation indexes to explore the optimal detection conditions. The methodology verification of specificity,linear range,precision and durability was conducted in accordance with the general principles of Chinese Pharmacopoeia(Volume Ⅳ,2020 edition)9101.Results The results of all the evaluation indexes were good when the elution ratio of organic phase was30% ～ 95%,the detection temperature was 35 ℃,the sample volume was 3 μg,and the elution time of 95% organic phase was 15 min. The method had the linear correlation coefficient(R2)of 0. 998 5,the linear range of 1. 8 ～ 4. 2 μg,the reproducibility RSD of 0. 01%,and the intermediate precision RSD of 0. 16%,with good durability under slight changes of column temperature and flow rate.Conclusion The reversed phase high-performance liquid chromatography method for the purity determination of recombinant Mtb Ag85b protein stock solution was developed,which has good specificity,precision and durability,and can be used for the quality control of recombinant Mtb Ag85b protein stock solution.

Objective To discuss the early diagnosis of severe perioperative biliary infection complicated with DIC and efficacy of comprehensive treatment for it. Methods Comprehensive treat-ment was conducted in 21 patients who had intensive biliary infection complicated with DIC. The he-moglutination function was monitored before and after treatment. Results Of the 21 patients, 19 were cured and 2 dead. The curative rated was 90.5 %. The hemorrhage tendency was controlled, abdomi-nal bleeding stopped, prothrombin time (PT) obviously reduced, content of fibrinogen (Fbg)in-creased, level of D-dimer and FDPs obviously dropped. Conclusion The patients who have biliary in-fection in the perioperative period are easy to suffer from DIC and the mortality is high. Early diagno-sis and comprehensive therapy, especially the application of the cryofibrinogen and heparin, are very important.

Acute radiation dermatitis is one of the most common toxicities of radiotherapy in patients with nasopharyngeal carcinoma. It leads to a series of symptoms such as erythema, desquamation and necrosis, which severely affect the quality of life. No concensus has been achieved on the standard prevention and treatment. In this article, literature review was performed in the prevention and treatment of acute radiation dermatitis in patients with nasopharyngeal carcinoma and relevant problems and prospects were proposed, aiming to provide certain reference for clinical trial and scientific research.

Objective To investigate the clinical value of high frequency ultrasonography with E-flow imaging in the evaluation of fingertip's microcirculation changes in patients with systemic sclerosis(SSc).Methods Twenty-four SSc patients and 29 healthy subjects were involved.High frequency ultrasonography with E-flow imaging was used to observe the configuration and distribution of digital arteries in the last segment of left and right middle finger.Peak systolic velocity (PSV),end diastolic velocity (EDV),mean velocity(MV),vascular resistance index (RI) and pulsatility index(PI) of digital palmar propria arteries,nail bed arteries and finger ventral arteries were measured.Results In control group,rich blood supply was revealed within the fingertips.Digital palmar propria arteries,nail bed arteries and finger ventral arteries and their small branches were displayed clearly and continuously by E-flow imaging.While in SSc patients,the definition and continuity of fingertip's small vascular flow images were not as good as that in the control group,with the distribution of blood flow markedly reduced.compared with the control group,PSV,EDV and MV of digital palmar propria arteries,nail bed arteries,finger ventral arteries were decreased in SSc group(P＜0.01),but both RI and PI were increased(P＜0.05).There were no statistically significant differences between left and right fingertip's arteries index in normal control group (P＞0.05).But PSV,EDV and MV of left digital palmar propria arteries in SSc group were higher than that of the right(P＜0.05),whose differences bear statistic significance.Conclusions High frequency ultrasonography with E-flow imaging is sensitive and reliable to reflect fingertip's microcirculation changes and provide a new method to assess microvascular changes in SSc patients.

Objective Pemetrexed and β-elemene can inhibit the growth of tumor cells .This study was to investigate the effect of pemetrexed combined with β-elemene on the proliferation and apoptosis of cervical cancer HeLa cells. Methods Cervical cancer HeLa cells were treated with pemetrexed at the concentrations of 38, 76, 152, 228, and 304μg/mL, and at 24 and 48 hours of treatment subjected to MTT for detection of their proliferation .The experiment included four groups , with the cells treated with β-elemene ( 125μg/mL) , pemetrexed ( 76 μg/mL ) , β-elemene ( 125 μg/mL ) +pemetrexed (76μg/mL), and nothing (blank control) for 24 hours, followed by determination of their proliferation and apoptosis by MTT and flow cytometry, respectively. Results Pemetrexed at 38, 76, 152 and 228μg/mL inhibited the proliferation of the HeLa cells in a concentration-dependent manner, with the inhibition rates of (7.24 ±3.78), (7.94 ±4.37), (11.10 ±2.86) and (15.88 ± 3.38)%at 24 hours, and (16.69 ±0.95), (22.54 ±1.53), (24.48 ±0.92) and (25.54 ±3.61)%at 48 hours, both with statis-tically significant differences between any two groups (P<0.05).Significant differences were also found in the proliferation rate of the same concentration of pemetrexed at the two time points (P<0.05).The combination of pemetrexed and β-elemene showed an inhibi-tion rate of (49.95 ±5.76)%at 24 hours, remarkably higher than (24.36 ±5.59)%in theβ-elemene group and (10.69 ±1.37)%in the pemetrexed group (P<0.01). Conclusion Pemetrexed combined with β-elemene can significantly inhibit the proliferation and synergistically accelerate the apoptosis of HeLa cells .

Objective:To study the relationship between IL-1β and TNFα expression and cell apoptosis in gingival tissue of the subjects with diabetes associated periodontitis(DAP).Methods:20 cases of DAP(group DAP) and 20 cases of health controls(group H)were included.The cell apoptosis and the ultrastructural changes in gingival tissue were observed by Tunnel staining and transmission electron microscope (TEM).IL-1β and TNFα expression in gingival tissue were detected by immunohistochemical staining.SBI,GI,PD and AL of the subjects were measured.The relationship between the level of IL-1β,TNFα and the cell appotosis was analyzed.Results:Apoptosis was obvious in prickle cells and basal cells of gingival tissue of DAP group.The percentage of apoptosis cells of DAP group was significantly higher than that of group H(P＜0.01).The expression of IL-1β and TNFα in group DAP was significant higher than that of group H (P＜0.01),and the mainly positive expression cells were macrophages,plasmocytes and lymphocytes.Conclusion:IL-1β and TNFα play a role in cell apoptosis in the gingival tissue of the patients with DAP.

Objective To investigate the significance of computed tomography(CT)and 3.0 T magnetic resonance imaging(MRI)in intensity-modulated radiotherapy(IMRT)for esophageal carcinoma. Methods Thirty-five patients newly diagnosed with esophageal carcinoma who received radical radiotherapy in our hospital from November 2013 to April 2015 were enrolled as subjects. Target volume was delineated on the CT images and MRI images(T2-weighted and diffusion-weighted fusion images). The MRI-and CT-based IMRT plans were designed using the same dose prescription and dose constraints for organs at risk(OAR). The target volume,prescribed dose,and doses for OAR were compared between the two plans. Results In the two plans, dose distribution and planning parameters met the clinical requirement. The length of lesion,gross tumor volume (GTV),and planning target volume(PTV)defined by 3.0 T MRI were significantly smaller than those defined by CT(P=0.00,0.03,0.03). There were no significant differences in the D 2%,D 98%,D 50%,homogeneity index,or conformity index for primary GTV(PGTV)and PTV-PGTV between the two plans(all P>0.05). Compared with the CT-based plan,the 3.0 T MRI-based plan had a significantly smaller mean dose to the lungs and an insignificantly smaller actual dose to the lungs(P=0.00;P>0.05).There were no significant differences in maximum doses tolerated by the spinal cord or heart between the two plans. Conclusions In terms of target volume delineation and dosimetric parameters, both CT-and 3.0 T MRI-based plans meet the clinical requirement. The 3.0 T MRI-based plan may provide potential benefits for some OAR due to a smaller target volume compared with the CT-based plan.

Objective:To explore the efficacy of digital and 3D printing technologies on design of superficial iliac circumflex artery flap for coverage the donor site of anterolateral thigh flap(ALTF).Methods:Clinical data of 8 patients were studied retrospectively for treatment of soft tissue defects of hand in the Department of Hand Surgery, Wuxi NO.9 People's Hospital Affiliated to Soochow University, from April 2017 to October 2021. The patients were 6 males and 2 females, aged from 29 to 59 years(mean, 45.8 years). Cause of injury: 3 patients were crushed, 2 by hot pressing, and 3 by machine strangulation. Site of injury included: 5 cases were dorsal hand defects and 3 cases were palm defects. All the wounds were contaminated to varying degrees with soft tissue defects. The areas of soft tissue defect ranged from 11 cm×10 cm to 22 cm×14 cm. Four patients had combined injuries of open fracture of metacarpals and phalanges and 3 with tendon defects. All wounds were repaired by free ALTF transplantation. And the donor sites in the thigh were repaired by superficial iliac circumflex artery flaps. The secondary wounds caused by flap harvesting on abdominal wall were closed directly. The targeted perforator vessels were detected preoperatively by CTA combined with CDU. 3D printed models of the affected hand were obtained before operation for individualised repairs according to the shape and area of the wounds. After the operation, all patients entered scheduled follow-ups at the outpatient clinic and via internet by observing the flap shape and testing the recovery of sensory and movement of adjacent joint.Results:The shapes and sizes of the wounds and the flaps were found basically in accordance with those in the preoperative simulative designs. All flaps in 8 patients survived and the wounds healed completely. All patients entered follow-ups for 8 to 24(average, 17.5) months. The donor thighs presented good appearance and colour, pliability without bloating. The range of motion of the hips and knees was not affected. Only linear scars remained in the abdominal donor sites, with natural colour and appearance.Conclusion:Digital and 3D printing technologies in preoperative design of flaps can help to locate the perforator vessels intraoperatively and guide the individualised design of the flaps with improved operation efficiency and satisfactory appearance of the flaps.

Objective:To investigate the changes in adaptive phenotypes of Yersinia pestis ( Yp) during successive passages in macrophages. Methods:A Yp strain of 201-MI was induced by 50 successive passages of Yp 201 strain in Raw264.7 cells. Phenotypic characteristics of 201 and 201-MI strains were compared by analyzing their survival rates in macrophages, growth curves, biofilm formation abilities, acid and hydrogen peroxide-stress tolerance, and virulence to mammal cells (Raw264.7 and HeLa cells) and mice. Results:Comparing with 201 strain, 201-MI strain showed various phenotypic changes, including higher survival rate in Raw264.7 cells, faster growth in iron-deficient medium, higher tolerance to acid and hydrogen peroxide, decreased biofilm formation ability, and less damages to Raw264.7 and HeLa cells. More-over, 201-MI strain showed decreased virulence to mice in both subcutaneous and intraperitoneal challenges. Preliminary comparative genomics analysis revealed some indel and nonsense mutations in 201-MI strain, which might account for its phenotype changes.Conclusions:After successive passages in macrophages, Yp showed some phenotypic changes, which might reflect its adaptive evolution under the pressure of macrophages. Detailed multi-omics analysis would be of great help to understand the underlying genetic mechanisms of these changes, and the related Yp-macrophage interaction processes as well.

AIM:To explore the key points of per-sonalized treatment and medication monitoring by clinical pharmacists in the perioperative period of moyamoya disease patients with endometriosis.METHODS:This study showed a moyamoya disease patient with endometriosis who accepting dual an-tiplatelet therapy and how to adjust medication during the perioperative period.Besides,monitor-ing the use proton pump inhibitors in the perioper-ative period and the formulating a plan of DIE post-operative recurrence prevention are also our objec-tives.RESULTS:Through individualized assessment of the patient,a personalized medication adjust-ments for dual antiplatelet therapy was deter-mined.Individualized perioperative proton pump inhibitor regimen and postoperative prevention plan for endometriosis recurrence were developed.CONCLUSION:The development of individualized medication regimens and medication monitoring is the breakthrough point for clinical pharmacists to carry out pharmaceutical services.