With the advantage of clear target and little side effect, antibody drug has attracted widely attention of worldwide pharmaceutical companies. However, large scale mammalian cell culture and antibody quality analysis are the bottlenecks of antibody drug industrialization in China. Especially due to the significant effect of cell culture conditions on antibody heterogeneity. Therefore, it is extremely urgent to optimize cell culture conditions to favor the demands of antibody drug development. This review summarized the most recent advances in the effect of cell culture conditions on antibody quality, followed by addressing the key issues that might be strategically important for domestic antibody drug development.
Animals ; Antibodies, Monoclonal ; biosynthesis ; isolation & purification ; Antibody Formation ; Cell Culture Techniques ; methods ; trends ; Genetic Heterogeneity ; Protein Stability ; Quality Control

BACKGROUND:Systemic lupus erythematosus is a kind of heterogeneous disease, and the difference of clinical features may also be the risk factors of osteonecrosis besides of treatment with glucocorticoids according to the literature. However, it remains controversial on the risk factors of osteonecrosis in systemic lupus erythematosus patients.
OBJECTIVE:To systematical y review the major risk factors of osteonecrosis in the Chinese patients with systemic lupus erythematosus.
METHODS:The CNKI database, CBMdisc database and Wanfang database were retrieved for the published case-control study literatures on the risk factors of osteonecrosis in the Chinese patients with systemic lupus erythematosus. The literatures met the inclusion and exclusion criteria were included, and a Meta-analysis was conducted by RevMan 5.0 and Stata software. Then, the pooled odd ratio and 95%confidence interval of each risk factor were calculated.
RESULTS AND CONCLUSION:Ten case-control study literatures were included involving 332 cases in the case group and 986 cases in the control group. The pooled odd ratio and 95%confidence interval of each risk factor of osteonecrosis in the Chinese patients with systemic lupus erythematosus were as fol ows:Raynaud’s
phenomenon 3.28(1.69-6.38), dental ulcer 2.95(2.13-4.09), renal involvement 1.21(0.83-1.74), vasculitis 5.64(2.84-11.21), hyperlipidemia 5.11(3.10-8.42), anti-phospholipid antibody 2.32(1.49-3.61) and joints involvement
2.02(1.33-3.07). It has been clear that the glucocorticoids is an independent risk factor of osteonecrosis in the patients with systemic lupus erythematosus. However, it is not the one and only risk factor. The fol owing risk factors of
vasculitis, hyperlipidemia, Raynaud’s phenomenon, dental ulcer, positive anti-phospholipid antibody and joints involvement are the risk factors of osteonecrosis in the patients with systemic lupus erythematosus.

Objective:To optimize the formula of ropivacaine hydrochloride transdermal gel. Methods:The steady transdermal rate and cumulative transdermal percentage in 24 h of ropivacaine hydrochloride gel were used as the indices, an orthogonal design was applied to select the optimal formula, and Design Expert 8. 0. 5. 0 software was used to analyze the results. Results: The optimal formula con-tained 2% carbomer, 10% propylene-glycol and 5% Azone. The steady transdermal rate of the optimal formula was 0. 6951 mg·h-1 · cm-2 . The cumulative transdermal percentage in 24 h of the optimal formula was 91. 04%, which was 22. 79% higher than that of ropiva-caine hydrochloride solution with the same concentration. Design Expert 8. 0. 5. 0 software could predict the steady transdermal rate and cumulative transdermal percentage in 24 h of the optimal formula. Conclusion: The preparation design is reasonable, and the gel has promising properties, which is suitable for skin local application.

Objective To investigate effects of total flavonoids of litchi (TFL) on the proliferation of rat hepatic stellate cells (HSC-T6) in comparison with western medicine rosiglitazone, and to explore the mechanism of anti hepatic fibrosis of TFL. Methods Effect of TFL on proliferation of HSC-T6 was examined by MTT. The expression of peroxisome proliferator-activated receptor-γ (PPAR-γ) mRNA, connective tissue growth factor (CTGF) mRNA in HSC-T6 cells exposured were examined by real-time quantitative PCR. Effects on HSC-T6 CTGF protein from TFL and rosiglitazone were detected by Western bloting. Results The expression of PPAR-γ mRNA was upregulated and the expression of CTGF mRNA and protein was downregulated after exposure to TFL and rosiglitazone for 72 hours. And the effect of TFL increased with the increase of concentration. Conclusion TFL can inhibit the proliferation of HSC-T6 and antagonizing liver fibrosis. This mechanism may be associated with the upregulation of PPAR-γ expression and the downregulation of CTGF expression.

Objective:To investigate the effects of three herb extracts on cell proliferation and collagen synthesis in cultured hu-man dermal fibroblasts. Methods:The human dermal fibroblasts were isolated and cultured with tissue explant culture technique. Im-munohistochemical staining was performed to observe vimentin in fibroblasts. The proliferation of fibroblasts was determined by MTT as-say. Hydroxyproline in the cultural medium was determined by a digestion method. Results:The cells were typical fibroblasts, and the immunohistochemistry staining of vimentin in the cells was positive. The three herb extracts could promote the proliferation of the fibro-blasts at appropriate concentrations in a time-dependent manner. After the 72-hour culture in the medium, 0. 312 5-5. 000 0 mg·ml-1 Aloe vera gel extract, 0. 156 3-2. 500 0 mg·ml-1 Bletilla striata extract and 0. 075 0 mg·ml-1 salvianolate showed significant effects on the proliferation of fibroblasts (P<0. 05 or P<0. 01 vs the blank group). Compared with that in the blank group, the amount of hydroxyproline in the three herb extracts cultural medium was also increased (P<0. 05 or P<0. 01). The effect of salvianolate on the cell proliferation was closely related with time and concentration, and the high concentration group could inhibit the growth of fibroblasts as the extension of time. Conclusion:Aloe vera gel extract, bletilla striata extract and salvianolate can promote the proliferation of fi-broblasts and the production of hydroxyproline, which may contribute to the wound healing.

Objective:To optimize the formula of amlexanox nasal thermosensitive gel spray and establish the quality control meth-od. Methods:Amlexanox nasal thermosensitive gel spray was prepared by a cold dissolving method, and poloxamer 407 (P407) and poloxamer 188 (P188) were used as the carrier materials. Central composite design-response surface methodology was used to optimize the formula with the amount of P407 and P188 as the influencing factors and the gel temperature and the viscosity before gelling as the indices. The content of amlexanox was determined by HPLC. According to Chinese Pharmacopoeia (2010 edition), the other indices of the preparation such as appearance, pH, viscosity, content, total spray times of each bottle and the content of each spray were deter-mined as well. Results:The optimum ratio of P407 and P188 was 17. 0% and 0. 9%,respectively . The average recovery of amlexanox was 98. 8% and RSD was 1. 7%(n=9). The quality of 3 batches of amlexanox nasal thermosensitive gel spray met the related require-ments. Conclusion:The formula and preparation process of amlexanox nasal thermosensitive gel spray are reasonable and feasible with controllable quality, which is worthy of further research.

OBJECTIVETo explore combination rules of Chinese herbal prescriptions from effective cases for treatment of unstable angina (UA).

METHODSPrescription data from 156 UA patients effectively treated at Cardiovascular Diseases Centre of Xiyuan Hospital were analyzed using complex network method.

RESULTSAccording to multi-scale analysis of backbone network and pointwise mutual information analysis, core prescriptions from the 156 UA patients were presented as follows: Rhizoma Ligustici wallichii, Radix Paeoniae rubra, Radix Codonopsis, Rhizoma Pinelliae, poria, and Angelica sinensis. Meanwhile, core couplet medicines for these patients covered Rhizoma Ligustici wallichii and Radix paeoniaerubra, Angelica sinensis and Rhizoma Ligustici wallichii, Radix Codonopsis and Rhizoma Ligustici wallichii, Rhizoma Ligustici wallichii and Rhizoma Pinelliae, Rhizoma Atractylodis Macrocephalae and poriacocos, Bulbus Alli Macrostemi and Rhizoma Pinelliae. Among different primary symptoms, there was slightly difference in core prescriptions.

CONCLUSIONThe core prescriptions for the treatment of UA include blood-activating drug, phlem-resolving drugs. As an exploration of combination rules of Chinese herbal prescriptions in treating UA based on complex network, it can be used as a reference for further researches.

Angelica sinensis ; Angina, Unstable ; drug therapy ; Drugs, Chinese Herbal ; standards ; therapeutic use ; Humans ; Pinellia ; Plant Roots ; Practice Guidelines as Topic ; Prescriptions ; standards

Objective To investigate the inhibitory effects of overexpression of PTEN on renal epithelial-mesenchymal trans-differentiation induced by TGF-β1,and the signaling transduction mechanism.Methods HKC cells were transfected with GFP-PTEN via lipofectAMINE2000.The efficiency of transfection was detected by fluorescence microscope.The expression of PTEN protein and mRNA in the translected cells was detected by Western blot and RT-PCR respectively.The experiment was divided into four groups:normal group,TGF-β1 stimulation group,GFP-PTEN+TGF-β1 group and empty vector+TGF-β1 group.The expression of E-cadherin,a-SMA,Akt and p-Akt was detected by Western blot.Results Most ceils transfected with GFP-PTEN expressed GFP.The expression of PTEN protein and mRNA was strongly increased when HKC cells were transfected with GFP-PTEN(all P<0.05).In both TGF-β1 stimulation group and empty vector+TGF-β1 group,the expression level of E-cadherin was lower(all P<0.05),while that of p-Akt and a-SMA was higher than in normal group(both P<0.05).The expression level of p-Akt and a-SMA in GFP-PTEN+TGF-β1 group was Iower(both P<0.05),while that of E-cadherin was higher than in TGF-β1 stimulation group and empty vector+TGF-β1 group(both P<0.05).The expression of Akt was similar in the four groups.Conclusion Overexpression of PTEN can inhibit renal epithelial-mesenehymal trans-differentiation induced by TGF-β1 through suppressing the activation of PI3K/Akt signal pathway.

Objective To investigate the impact of sputum suction under laryngoscope in stroke patients on the occurrence and prognosis of ischemic stroke-associated pneumonia (SAP).Methods A total of 127 cases with acute ischemic stroke (AIS) with Glasgow Coma Scale 6-12 scores,non-artificial airway and need for intensive care were enrolled.According to the suction practice,the cases were divided into laryngoscope euthyphoria group (practice group,PG,n =59),and conventional oral or nasal sputum suction group (control group,CG,n =68).The PG was perspectively studied,while the CG were analyzed retrospectively.All patients were cared with the Guidelines for the management of adult with hospitalacquired ventilator-associated and healthcare-associated pneumonia set by American Thoracic Society/Infectious Diseases Society of America in 2005 and the Guidelines for the early management of patients with acute ischemic stroke set by American Heart Association/American Stroke Association in 2013 including patting on the back of patients in prone position routinely in order to facilitating expectoration,phlegm elimination with physical vibration and apophlegmatic agent application during their stay in ICU.Incidences of SAP and 28-day mortality were calculated.Levels of inflammatory markers (white blood cell,WBC;highsensitive C-reaction protein;procalcitonin,PCT) in serum were detected.Adverse events related to laryngoscopy were documented.Results There were no significant differences in incidences of SAP and 28-day mortality between PG and CG groups (32.20％ vs.41.18％,P ＞ 0.05;47.37％ vs.53.57,P ＞ 0.05,respectively).However,there was statistically significant difference (P ＜ 0.05) in the ICU stay length and inflammatory markers,and the sputum suction process in PG did decrease the incidence of artificial airway establishment and mechanical ventilation (18.64％ vs.35.29％,P ＜ 0.05),and the laryngoscope euthyphoria method was proved to be a protective factor (HR =0.515;95％ CI 0.281-0.057,P =0.038).WBC and PCT were lower in PG group than in CG group (P ＜ 0.05).Conclusion Sputum suction under laryngoscope can reduce the probability of artificial airway establishment and invasive ventilator employment in patients with AIS,shortening ICU stay length and down-regulating levels of WBC and PCT.

Objective To explore prognosis and remission-related factors in lupus nephritis (LN) patients.Methods Patients diagnosed as LN by renal biopsy in Tongji Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology between Jan 1,2011 and July 31,2016 were enrolled.All related baseline clinical data was recorded and regular follow-up was performed.Kaplan-Meier curves was used to analyze partial remission and complete remission rates.Log-rank test was performed to compare remission rates of patients with nephrotic-range proteinuria (24-hour proteinuria≥3.5 g) and without nephrotic-range proteinuria (24-hour proteinuria＜3.5 g).Univariate and muhivariate Cox regression analyses were performed to evaluate the remission-related factors in different periods.Results A total of 115 patients,with 88.7％ female and (31.5±9.5)years mean age,were followed up for up to 5 years.During follow-up period 2 patients died and 1 dialyzed.The 6-,12-,24-and 36-and 48-month renal partial remission and complete remission rates were 33.3％,58.2％,71.5％,84.0％,89.6％,and 18.9％,40.5％,67.3％,79.4％,87.0％,respectively.Patients without nephrotic-range proteinuria had higher complete remission than patients with nephrotic -range proteinuria (HR=2.01,95％CI 1.15-3.34,P=0.014),but there was no difference in their partial remission (HR=1.33,95％ CI 0.74-2.43,P=0.341).Multivariate Cox regression model indicated that every 1 g/L increase in baseline level of serum albumin was associated with increased 8％ and 9％ risk,respectively,in partial remission (HR=1.08,95％CI 1.01-1.15,P=0.024) and complete remission (HR=1.09,95％CI 1.01-1.07,P=0.038).Conclusions Around half of LN patients reach remission during 1 year.Patients without nephrotic-range proteinuria have higher complete remission,and serum albumin is a remission-related factors.