As the need for antibody production rises, there is an urgent need to lower the costs and enhance the efficiency of the separation process. Currently, the chromatographic media used for antibody separation and purification often focus on individual properties of antibodies, such as affinity, hydrophobicity, and charge, leading to issues like low purification efficiency or inadequate adsorption capacity. To address this, an electrostatically coupled polypeptide affinity medium (FD7-3, 5-diaminobenzoic acid n-sepharose, FD7-DA-Sepharose) was developed for rapid purification of antibodies from cell culture supernatant. This medium utilized 3, 5-diaminobenzoic acid as a spacer to attach the heptapeptide-affinity ligand (FYEILHD, FD7) to agarose microspheres. Antibodies could be adsorbed through charge interactions with the carboxyl functional group of the FD7-DA-Sepharose spacer, while FD7 enhanced electrostatic coupling and affinity adsorption through synergistic effects, significantly increasing the adsorption capacity while maintaining the affinity and specificity. The influences of pH and ionic strength on adsorption capacity were investigated with human immunoglobulin as a model protein. The static adsorption capacity (Q_m) of FD7-DA-Sepharose in the solution of pH 6.0 reached 67.73 mg/mL, representing a 52.68% increase compared with that (44.36 mg/mL) of the commercial Protein A affinity medium. Furthermore, the elution conditions for FD7-DA- Sepharose were mild (20 mmol/L PB, 0.5 mol/L NaCl, pH 6.0), in contrast to the harsh acidic elution (pH 2.7-3.6) typically associated with Protein A, which can damage antibody integrity. The FD7-DA-Sepharose medium was then employed to purify antibodies from cell culture supernatant, achieving the yield of 94.8% and the purity of 98.4%. The secondary structure of the purified antibody was determined by circular dichroism spectroscopy. The results demonstrated that FD7-DA-Sepharose enabled efficient purification of antibodies from cell culture supernatant, which provided a cost-effective solution (approximately one-third the price of commercial Protein A affinity medium) with gentle elution conditions that preserve the natural conformation of antibodies. This approach paves a novel, economical, and efficient way for the separation and purification of antibodies from cell culture supernatant.
Chromatography, Affinity/methods* ; Static Electricity ; Humans ; Sepharose/analogs & derivatives* ; Peptides/chemistry* ; Adsorption ; Antibodies/isolation & purification*

OBJECTIVE To establish a quantitative analysis model for the contents of tussilagone， moisture， ethanol-soluble extract and total ash in Farfarae Flos based on near-infrared spectroscopy （NIRS）， providing a new idea for the rapid quality evaluation of Farfarae Flos and its preparations. METHODS Referring to the 2020 edition of the Chinese Pharmacopoeia， the contents of the main quality control indexes tussilagone， moisture， ethanol-soluble extract and total ash in 130 batches of Farfarae Flos from 19 producing areas were determined by HPLC， drying method， hot dip method and ash assay， respectively. The NIRS data information of the medicinal herbs of Farfarae Flos was collected， and then NIRS combined with the partial least squares method was used to establish the individual quantitative analysis models of the above quality control indexes in the samples， and the predictive model of the NIRS content was obtained after sample validation with validation set. RESULTS The range for the contents of tussilagone， moisture， ethanol-soluble extract and total ash in 130 batches of Farfarae Flos were 0.051 4%-0.103 5%， 7.75%-10.93%， 20.17%-31.12%， and 7.68%-12.10%， respectively. The internal cross-validation coefficients of determination （R2） of the established models for the quantitative analysis of tussilagone， moisture， ethanol-soluble extract and total ash in Farfarae Flos were 0.985 8， 0.968 4， 0.973 4， 0.988 0， respectively； the root mean square errors of calibration （RMSEC） were 0.001 54， 0.187， 0.478， 0.127， respectively； the root mean square errors of prediction （RMSEP） were 0.001 81， 0.212， 0.543， 0.149， respectively； RMSEP/RMSEC were 1.175 3， 1.133 7， 1.136 0 and 1.173 2， respectively， which were all within a reasonable range （1＜RMSEP/RMSEC≤1.2）. The mean absolute errors between the true and model-predicted values of the above four quality control indexes in the validation set of samples were －0.000 36， 0.061 43， 0.144 00， and 0.010 43， respectively，and the mean predicted recoveries were 99.65%， 100.72%，100.66%， and 100.15%， respectively. CONCLUSIONS The established NIRS quantitative analysis model has high stability and reliable results， which can be used for the rapid batch prediction of the content of relevant quality control indexes in Farfarae Flos.

Objective    To evaluate the safety, feasibility and short-term outcomes of single-direction gastric mobilization under 3D-laparoscopy in minimally invasive esophagectomy for the treatment of esophageal cancer. Methods    From February 2018 to December 2019, 118 consecutive patients who underwent minimally invasive McKeown esophagectomy for esophageal squamous cell carcinoma in our hospital were included. There were 94 males and 24 females with an average age of 53.7 (41–77) years. They were divided into two groups based on the methods of gastric mobilization: a traditional dissociation (TD) group (n=55) and a single-direction mobilization (MD) group (n=63). The clinical data of the two groups were compared. Results    Enbloc resection and a negative resection margin were obtained in all patients. There was no postoperative mortality or incision complication. The rate of postoperative complications was 22.9%. There was no significant difference in the spleen injury, gastric injury, conversion to open surgery, abdominal reoperation as well as cervical anastomotic leakage between the two groups (P>0.05). It took significantly less time in the MD group compared with the TD group (P<0.05). There was an obvious statistical difference in the incidence of gastric mobilization related complications between the MD group (1.6%, 1/63) and TD group (12.7%, 7/55, P<0.05). Conclusion    Application of single-direction gastric mobilization under 3D-laparoscopy in minimally invasive esophagectomy for the treatment of esophageal cancer is safe and easy to perform with a satisfactory short-term outcome.

Objective:To observe the clinical efficacy of Danzhi Sangqin decoction in the treatment of premenstrual acne with liver and lung stagnation.Methods:Sixty patients with liver and lung stagnation premenstrual acne diagnosed and treated in the Department of Dermatology of Shaoxing Central Hospital from July 2016 to November 2018 were selected in the research.They were divided into the control group( n=28) and treatment group( n=32) by random number table method.The treatment group was treated with self-made Danzhi Sangqin decoction, and the control group was orally treated with isotretinoin capsules and vitamin B6.The two groups were treated for 1 week.The clinical efficacy and the incidence of adverse reactions were compared between the two groups. Results:In the treatment group, 18 cases were markedly effective, 11 cases were effective, and 3 cases were ineffective, and the total effective rate was 90.6%(29/32). In the control group, 16 cases were significantly effective, 10 cases were effective, and 2 cases were ineffective, and the total effective rate was 92.9%(26/28). There was no statistically significant difference in the total effective rate between the two groups( P>0.05). The incidence of adverse reactions in the treatment group was 3.1%(1/32), which was significantly lower than that in the control group [21.4%(6/28)], the difference was statistically significant(χ 2=6.281, P<0.05). Conclusion:The clinical efficacy of Danzhi Sangqin decoction in the treatment of premenstrual acne with liver and lung stagnation is similar to that of isotretinoin capsules, but the incidence of adverse reactions is low.

Objective:To study the effects of compound schisandra extracts (CSE) (schisandra,astragalus,acanthopanax,and rhodiola)on the exhaustive swimming time and the levels of blood urea nitrogen(BUN),serum lactic acid(LD),liver glycogen and muscle glycogene,superoxide dismutase(SOD)activity,and malonaldehyde(MDA) level in the mice and to charify its anti-fatigue effect and the mechanism.Methods:Eighty male ICR mice were randomly divided into blank control group,50 mg·kg-1 CSE group,100 mg·kg-1 CSE group,and 200 mg·kg-1CSE group;there were 20 mice in each group.The mice were administered orally for 30 d.Then 10 mice were randomly selected for exhaustive swimming test in each group and the exhaustive swimming time of the mice was recorded.The remaining 10 mice in each group were used for 90 min swimming,then all the mice were sacrificed and the blood and tissue samples were taken for the measurement of the levels of BUN,LD,liver glycogen and muscle glycogen,the SOD activity and MDA level;the total inhibitory rate of oxidation of CSE in vitro was determined by linoleic acid-ferric thiocyanate method.Results:Compared with blank control group,the exhaustive swimming time of the mice in 50,100,and 200 mg·kg-1 CSE groups were significantly increased (P<0.01);the levels of BUN and LD of the mice in 100 and 200 mg·kg-1 CSE groups were significantly decreased (P<0.05 or P<0.01);the levels of liver glycogen and muscle glycogen of the mice in 100 and 200 mg·kg-1 groups were significantly increased(P<0.05 or P<0.01);whereas the SOD activities were significantly increased and the levels of MDA were decreased(P<0.05 or P<0.01).Compared with 100 mg·kg-1 CSE group,the levels of serum BUN and LD of the mice in 200 mg·kg-1 CSE group were decreased (P<0.01),and the levels of liver glycogen and muscle glycogen were increased(P<0.05).The total inhibitory rate of oxidation of 5 g·L-1CSE was 76.94%.Conclusion:CSE has an anti-fatigue effect and the mechanism may be related to anti-oxidation effect.