Objective:To systematically investigate the short-term efficacy of total knee arthroplasty in the treatment of osteoarthritis coupled with fixed patellar dislocation.Methods:A retrospective analysis was conducted on a cohort of 11 patients diagnosed with knee osteoarthritis and fixed patellar dislocation who underwent total knee arthroplasty at Panzhihua Central Hospital from January 2018 to October 2021. The cohort comprised 4 males and 7 females, aged 63.45±4.76 years (range, 56-70 years), all of whom underwent unilateral surgery. There were 5 left and 6 right knees, with a body mass index of 23.20±2.02 kg/m 2 (range, 20.8-27.6 kg/m 2) and a disease course of 12.63±4.81 years. According to the American Society of Anesthesiologists classification, 9 cases were categorized as grade II, and 2 cases as grade III. Recovery of patellar trajectory during total knee arthroplasty, using medial synovial flap transposition to repair lateral joint capsule. Preoperative and postoperative assessments included knee joint range of motion, Knee Society score (KSS), University of California Los Angeles (UCLA) score, and visual analogue scale (VAS). Results:All 11 patients were followed up for a period of 28.64±4.01 months (range, 24-36 months). Two patients exhibited subcutaneous fat liquefaction locally after surgery, which resolved following dressing changes. All wounds achieved primary healing. Two of them developed intramuscular vein thrombosis after surgery and were cured after anticoagulant treatment. The range of motion of the knee joint increased from 63.18°±17.07° before surgery to 104.55°±16.80° at the last follow-up, with a statistically significant difference ( t=14.041, P<0.001). The KSS score increased from 38.00±6.78 points to 80.91±5.65 points, with a statistically significant difference ( t=16.472, P<0.001). The UCLA score increased from 3.18±1.17 to 6.73±1.35, with a statistically significant difference ( t=9.694, P<0.001). The VAS decreased from 6.09±0.94 points to 2.32±0.64 points, with a statistically significant difference ( t=16.600, P<0.001). At the last follow-up, imaging examinations showed no cases of patellar subluxation or dislocation, no tearing or breakage of the knee extension device, and no infection or loosening around the prosthesis. Conclusion:Utilizing medial synovial flap transposition for repairing the lateral joint capsule proves to be an effective technique for key capsule repair. Total knee arthroplasty for osteoarthritis combined with fixed patellar dislocation demonstrates satisfactory early clinical outcomes.

Objective To analyze the relationship between angiotensin type Ⅱ 1 receptor autoantibodies (AT1-AA),blood uric acid and urea nitrogen with hypertensive disorder complicating pregnancy (HDCP). Methods A total of 125 patients with HDCP diagnosed in the obstetrics department of Tianjin Municipal Bao-di District People's Hospital from August 2021 to February 2023 were randomly selected as the HDCP group, and 55 healthy pregnant women admitted to this hospital during the same period were selected as the control group.The HDCP group was divided into the gestational hypertension subgroup (n=77) and preeclampsia group (n=48) according to the degree of disease progression.The levels of serum uric acid,blood urea nitro-gen and AT1-AA were detected and compared between the HDCP group and control group and between the gestational hypertension subgroup and preeclampsia subgroup.Results There were no statistically significant differences in the age,gestational time,D-dimer,serum calcium and serum creatinine between the control group and the HDCP group (P>0.05).The levels of peripheral blood AT1-AA,serum uric acid and blood u-rea nitrogen in the HDCP group were higher than those in the control group,and the differences were statisti-cally significant (P<0.05).The levels of peripheral blood AT1-AA,serum uric acid and blood urea nitrogen in the preeclampsia subgroup were significantly higher than those in the gestational hypertension subgroup, and the differences were statistically significant (P<0.05).The Spearman rank correlation coefficient analysis showed that the levels of peripheral blood AT1-AA (r=0.301),serum uric acid (r=0.380) and blood urea nitrogen (r=0.257) levels were positively correlated with the severity degree of HDCP (P<0.01).The area under the curve (AUC) of AT1-AA,serum uric acid and blood urea nitrogen detection alone and combined de-tection in the diagnosis of HDCP were 0.680,0.714,0.647 and 0.725,and the corresponding Youden coeffi-cients were 0.368,0.380,0.306 and 0.439,indicating that the value of combined testing was higher than that of single item detection in the diagnosis of HDCP.Conclusion AT1-AA,serum uric acid and blood urea nitro-gen are correlated with the occurrence and development of HDCP,and the 3-item combined detection has high-er value.

ObjectiveTo study the effects of Toddalia asiatica alcohol extract on autophagy and apoptosis of non-small cell lung cancer A549 cells， and to explore its possible mechanism. MethodA549 cells were cultured in vitro. Cell counting kit-8 （CCK-8） was used to detect the proliferation of A549 cells， and cell survival rate was calculated to screen the drug concentration. The apoptosis in each dose group and that after the use of 3-methyladenine （3-MA）， an autophagy inhibitor， were detected by flow cytometry combined with Annexin V-FITC/PI double staining. Western blot was used to detect the expression levels of apoptosis-related proteins such as B cell lymphocytoma-2（Bcl-2）， Bcl-2-associated X protein（Bax）， microtubule-associated protein 1 light chain 3 （LC3）， cleaved cysteinyl aspartate-specific protease-3 （cleaved Caspase-3）， activated poly （Adenosine diphosphate） ribonucleotide polymerase （cleaved PARP1）， PARP1， activated death activator （t-Bid）， Bid， and ubiquitin-binding protein p62 in each group and those after the use of 3-MA. ResultCompared with the conditions in the control group， the cell survival rate in 0.25 g·L^-1 group （P<0.05）， and 0.5， 1， 2， 4 g·L^-1 groups （P<0.01） was decreased after 24 h intervention. Additionally， the cell survival rate was reduced in a concentration-dependent manner at 48 h and it was less than 10% at 4 g·L^-1 （P<0.01）. Compared with the conditions in the control group， the total apoptosis rate in 0.5 g·L^-1 group was increased （P<0.05）， and the apoptosis rate in 1 and 2 g·L^-1 groups was also increased （P<0.01）. Compared with the 2 g·L^-1 group and 3-MA group， the 3-MA combined with T. asiatica alcohol extract had significantly decreased apoptosis rate （P<0.01）. Compared with the conditions in the control group， elevated expression of pro-apoptotic proteins cleaved PARP1， Bax and t-Bid in 1 and 2 g·L^-1 groups （P<0.05， P<0.01）， and reduced expression of Bid in the 2 g·L^-1 group （P<0.01） were found. Compared with the conditions in the control group， the expression of anti-apoptotic protein Bcl-2 （P<0.05， P<0.01） and the level of p62 （P<0.01） were down-regulated in 0.5， 1， 2 g·L^-1 groups， while the level of LC3 Ⅱ protein was up-regulated （P<0.01）， with certain concentration dependence. ConclusionT. asiatica alcohol extract could significantly inhibit the proliferation of A549 cells， which might be related to promoting autophagy and inducing apoptosis.

Single incision laparoscopic surgery (SILS) has many advantages than standard multiport laparoscopic cholecystectomy (MLC),such as small trauma,less postoperative pain,shorter hospital stay,good cosmetic effect,and so on.Especially the satisfying cosmetic result of no abdominal scar is more important.But the SILS is also faced with many difficulties.The operation time of SILS were significantly longer in duration than MLC.The safety and the technical feasibility were lower for the SILS whose operation complications are more than the MLC.The main reason is that the operation field don't exposureis sufficient and the formation of surgical operation triangle is not easy.In order to overcome these difficulties,scholars have used the auxiliary methods of exposing the operative field in various operation,including the penetration of abdomen wall retraction and intraperitonealretractionand abdomen wall retraction.This paper with review the advantages and disadvantages of the above methods aiming to affer more values for clinical doctors in opperating SILS who get more knonledge abont it.

The biological activity of ADCC by anti-CD20 monoclonal antibody was determined by BioGlo™ Luciferase Assay System using Jurkat/NFAT-luc+FcγRIIIa cell line as effector cell and WIL2-S cell line as target cell. The developed method was verified for specificity, precision and accuracy. Anti-CD20 monoclonal antibody showed a dose-response mode by the developed method, and the determination result complied with the following four-parameter equation: y = (A-D)/[1 + (X/C)(B)] + D. The optimized parameters of the method were determined including the antibodies diluted concentration (18,000 ng·mL(-1)), dilution rate (1:5), the ratio of effector cell and target cell (6:1), and induction time (6 h). The values of eight independent tests have passed a statistical test for curve regression analysis, linear or parallelism, which showed the method possessed good specificity. Four different dilute groups of recovery rates sample were determined for 3 times, and the result showed mean relative potencies of (44.39±3.93)%, (72.74±2.78)%, (128.28±7.01)% and (168.19±2.70)% respectively, with a variation coefficient of less than 10%, and the recoveries of (88.78±7.85)%, (96.99±3.70)%, (102.63±5.61)% and (112.12±1.80)% respectively. A novel reporter gene method for determination of biological activity of ADCC by anti-CD20 monoclonal antibody was successfully developed, which showed strong specificity, good reproducibility and high accuracy, and might be used routinely.

Objective To establish a HPLC method for simultaneous determination of seven active components in Fufang Danshen tablets and Fufang Danshen dripping pills. Methods These seven compounds were analyzed simultaneously with a Zorbax C18 column by gradient elution using acetonitrile-0. 1% phosphoric acid solution as mobile phase, the flow rate was 1 mL·min-1 and the detection wavelength was set at 203, 270 and 281 nm, respectively. Results All the seven components showed good linear relation between peak area and concentration of the test, and the average recoveries were between 95. 1%-100. 4%. Tanshinone ⅡA was not detected in samples of dropping pills. Conclusion The HPLC method to determine the components including tanshinone ⅡA, salvianolic acid B, propanoid acid, protocatechuic aldehyde, notoginsenoside R1 , ginsenoside Rg1 and ginsenoside Rb1 of the two different Danshen preparations has been established, and it has the advantages of simplicity, high precision, good repeatability, and can be used for the quality control of two kinds of Fufang Danshen preparations. The content of tanshinone Ⅱ A in Fufang Danshen tablet was distinctly higher than that of dropping pills.

Here we discuss whether N terminal sequencing is appropriate as one of the conventional control methods for monoclonal antibody products. We determined the N terminal sequences of two monoclonal antibody products targeting two antigens separately with both Edman degradation and mass peptide spectrometry. We also identified the characteristic peptide fragments with mass spectrometry. Furthermore, we analyzed their heterogeneity with ion exchange chromatography, capillary zone electrophoresis and Imaged Capillary Isoelectric Focusing. Edman degradation method showed that the N terminal 15 amino acids of heavy and light chains of the two monoclonal antibodies were identical. Peptide mass spectrometry demonstrated that T1 peptide fragments of heavy and light chains of the two antibodies were also the same. But in contrast, peptide mapping and the three analytical methods for heterogeneity analysis could effectively identify and differentiate the two antibodies. The N terminal sequences of two monoclonal antibodies are identical because the number of framework sequences of humanized or human monoclonal antibodies is relatively limited, so whether N terminal sequencing analysis could be regulated as one of the practical control methods should be carefully discussed. Our work also proves that the above analytical methods could combinatorially applied to the identification of monoclonal antibody products, and are more objective compared to N terminal sequencing.
Amino Acid Sequence ; Antibodies, Monoclonal ; isolation & purification ; Chromatography, Ion Exchange ; Humans ; Isoelectric Focusing ; Mass Spectrometry ; Peptide Mapping ; Peptides ; Sequence Analysis, Protein ; methods

Objective To analyze the differences of size and charge heterogeneities between origi-nal humanized anti-TNF-αantibody and four similar biotherapeutic products ( SBP ) .Methods The size exclusion chromatography ( SEC-HPLC ) and weak cation exchange chromatography ( WCX-HPLC ) were used to analyze the size and charge heterogeneities , respectively.Carboxypeptidase B (CpB) treatment was employed to analyze the source of charge heterogeneity of the antibody products .Results Four SBPs showed the same pattern with the originator in SEC-HPLC, and no significant difference with the percentage of mono-mer was observed .The percentages of the aggregates of SBP-3 and SBP-4 were a little higher than those of the originator .The charge distribution of SBPs was significantly different from the originator ′s, especially in the basic region .The results from the samples treated with CpB indicated that the difference of charge distri -bution in the basic region might be caused by the C-terminal lysine variants .Conclusion Four SBPs showed similar size heterogeneity with the originator , but significant differences with charge heterogeneity were observed among them .The study suggested that more attention should be paid to the charge heterogene -ity analysis of the biosimilar products .

Objective To compare the capability of capillary electrophoresis-sodium dodecyl sul-fate ( CE-SDS) and size exclusion-high performance liquid chromatography ( SE-HPLC) for analysis of size heterogeneity of monoclonal antibody products .Methods The size heterogeneity of one humanized anti-VEGF monoclonal antibody was analyzed by using non-reduced and reduced CE-SDS, and conventional , de-natured and denatured reduced SE-HPLC.Results The percentage of aggregates detected by non-reduced CE-SDS (0.82%±0.01%) was equal to that by using denatured SE-HPLC (1.05%±0.02%), but it was significantly lower than that by using conventional SE-HPLC analysis (5.08%±0.10%).With regard to fragments analyzed with non-reduced antibodies, its percentage was (7.12±0.04)% measured by non-re-duced CE-SDS analysis that was significantly higher than that by conventional SE -HPLC analysis (0.02%± 0.01%) and denatured SE-HPLC analysis (0.62%±0.01%).Using reduced antibodies , the percentage of fragments was (3.19±0.50)%tested by reduced CE-SDS analysis that was significantly higher than that by using denatured reduced SE-HPLC analysis (0.07%±0.01%).Conclusion Conventional SE-HPLC was more objective than CE-SDS for content analysis of aggregates , as both the covalent and non-covalent forms of aggregates could be detected .Non-reduced CE-SDS could demonstrate the content of clips , while reduced CE-SDS showed the degraded fragments .Therefore, CE-SDS had an advantage over conventional SE-HPLC for content analysis of fragments .The use of the two analytical methods in combination provided solid techni-cal supports for the quality control of size heterogeneity of monoclonal antibodies .

This paper reports the determination of the drug antibody ratio in an antibody-drug conjugate with two methods, i.e. LC-MS and UV/VIS, and to provide a reliable method to scientifically evaluate and effectively control the drug antibody ratio. Deglycosylated sample was analyzed with C4 column followed by MS, and the number of conjugated drugs in the antibody was determined by the molecular weight increase due to the addition of different number of drugs to the antibody, and then drug antibody ratio was calculated by weighted average of different number of drugs conjugated to the antibody. Optical density at 252 and 280 nm was measured with UV/VIS, and due to the difference of extinction coefficients between the antibody and the drug, the drug antibody ratio was calculated from linear equation with two unknowns. The drug antibody ratio was 3.21 and 3.25 respectively measured by the two methods, and the results were similar with the two methods. Our study indicated that both methods, LC-MS and UV/VIS, could be applied to the analysis of drug antibody ratio of the antibody drug conjugate.