Objective: To compare quality control (relative purity and specific activity) and process control [plasminogen (Pg) antigen recovery and potency recovery] indexes of samples before and after adding the Q chromatography step to the full chromatography process of human Pg, thereby determining whether the addition of this step could improve Pg recovery by affinity chromatography. Methods: A Q chromatography step was added before the Pg affinity chromatography in the original Pg chromatography process. The loading solution, flow through solution and eluate of Q chromatography and Pg affinity chromatography were collected. The potency of coagulation factor Ⅱ (FⅡ), Ⅶ (FⅦ), Ⅷ (FⅧ), Ⅸ (FⅨ), and Ⅹ(FⅩ) were detected by the coagulation method, the total protein content was detected by the BCA method, and the Pg potency was detected by the chromogenic substrate method. The content of specific plasma proteins was detected by immunoturbidimetry, the potency recovery of coagulation factors was calculated, and the flow direction of coagulation factors was analyzed. The recovery of different plasma protein antigens were calculated, and the distribution of impurity proteins was analyzed. The relative purity and specific activity of Pg, antigen content, and potency recovery in the target fractions were calculated and compared with the original process indicators, so as to determine the effect of adding Q chromatography on the original process. Furthermore, the reproducibility after process modification was assessed. Results: 100% of FⅡ, FⅩ, and FⅨ, 87.81% of FⅧ, and 40.44% of FⅦ in filtered plasma were removed by Q chromatography. The residual FⅦ (53.26%) and FⅧ (13.30%) in Q flow-through fraction were completely removed by Pg affinity chromatography. In both the original process (without Q-chromatography) and the modified process (with Q-chromatography), non-target plasma proteins mainly existed in the flow-through fraction of Pg affinity chromatography. The antigen recovery of IgM, ceruloplasmin (CER), and fibronectin (FNC) in Q-chromatography flow-through fraction were reduced. In contrast, antigen recovery of other plasma proteins [IgG, IgA, Pg, albumin (AlB), alpha-1-antitrypsin (AAT), and fibrinogen (Fg)] were all >90%, which were consistent with the protein composition and proportion in the original affinity chromatography loading solution. Compared with the recovery rate of Pg antigen in the original process (74.4%), the total recovery of Pg antigen in the modified process was significantly increased (89.97%). Compared with the recovery of IgG (97.48%) and Fg (95.32%) in the Pg affinity flows-through fraction of the original process, the modified process resulted in a slight reduction in the recovery of IgG (94.60%), while the recovery of Fg was not affected (95.05%). The potency recovery rate, specific activity, and relative purity of Pg after Q chromatography were 99.3%, 0.016 U/mg, and 0.15%. These values were the same as those of Pg affinity chromatography loading solution by the original process, indicating that introduction of Q chromatography did not affect subsequent Pg affinity chromatography. Compared with the recovery of Pg antigen in three batches of the original process (66.49±1.02)%, the recovery of Pg antigen in the affinity chromatography eluent of the modified process [five batches; (77.43±4.43)%] was significantly improved. Furthermore, the potency recovery was (86.80±4.28)%, the relative purity was (81.99±1.25)%, the specific activity was (8.679±1.073)U/mg, and the process was reproducible. Conclusion: The addition of Q chromatography could improve the recovery of Pg affinity chromatography in the full chromatography process.

By analyzing the medical insurance policy, the medical insurance audit business and the intelligent monitoring requirements put forward by the medical reform, the intelligent examination and approval database was established, and the system process reengineering was carried out in conjunction with HIS, LIS and electronic medical record systems. The intelligent medical insurance audit information system was successfully developed and implemented, which realized automatic review of the medical qualifications of trauma patients, automatic audit of blood product items, automatic verification of high-cost fees, automatic review of repeated hospitalizations within 10 days, and automatic update of discharge diagnosis, as well as drug( diagnosis/treatment) project limits( excessive) early warning and other core business functions. The system meets the actual needs of the quality and efficiency of data management in medical insurance audit, providing basic platform and data support for standardizing doctors′diagnosis and treatment behavior and avoiding hospital risks.

Objective: To investigate the clinical effect and safety of long-acting pegylated interferon-α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 μg/week) in the treatment of HBeAg-positive chronic hepatitis B (CHB) patients, with standard-dose Peg-IFN-α-2a as positive control. Methods: This study was a multicenter, randomized, open-label, and positive-controlled phase III clinical trial. Eligible HBeAg-positive CHB patients were screened out and randomized to Peg-IFN-α-2b (Y shape, 40 kD) trial group and Peg-IFN-α-2a control group at a ratio of 2:1. The course of treatment was 48 weeks and the patients were followed up for 24 weeks after drug withdrawal. Plasma samples were collected at screening, baseline, and 12, 24, 36, 48, 60, and 72 weeks for centralized detection. COBAS® Ampliprep/COBAS® TaqMan® HBV Test was used to measure HBV DNA level by quantitative real-time PCR. Electrochemiluminescence immunoassay with Elecsys kit was used to measure HBV markers (HBsAg, anti-HBs, HBeAg, anti-HBe). Adverse events were recorded in detail. The primary outcome measure was HBeAg seroconversion rate after the 24-week follow-up, and non-inferiority was also tested. The difference in HBeAg seroconversion rate after treatment between the trial group and the control group and two-sided confidence interval (CI) were calculated, and non-inferiority was demonstrated if the lower limit of 95% CI was > -10%. The t-test, chi-square test, or rank sum test was used according to the types and features of data. Results: A total of 855 HBeAg-positive CHB patients were enrolled and 820 of them received treatment (538 in the trial group and 282 in the control group). The data of the full analysis set showed that HBeAg seroconversion rate at week 72 was 27.32% in the trial group and 22.70% in the control group with a rate difference of 4.63% (95% CI -1.54% to 10.80%, P = 0.1493). The data of the per-protocol set showed that HBeAg seroconversion rate at week 72 was 30.75% in the trial group and 27.14% in the control group with a rate difference of 3.61% (95% CI -3.87% to 11.09%, P = 0.3436). 95% CI met the non-inferiority criteria, and the trial group was non-inferior to the control group. The two groups had similar incidence rates of adverse events, serious adverse events, and common adverse events. Conclusion In Peg-IFN-α regimen for HBeAg-positive CHB patients, the new drug Peg-IFN-α-2b (Y shape, 40 kD) has comparable effect and safety to the control drug Peg-IFN-α-2a.

Objective To analyze the rules for acupoint selection of warming acupuncture and moxibustion in clinical treatment of knee osteoarthritis based on data mining technology. Methods CNKI, Wanfang Data, CBM, VIP, PubMed and Cochrane Library were searched for relevant literature about warming acupuncture and moxibustion in treatment of knee osteoarthritis. Database was built according to the search results. Association rules were used to analyze the rules for acupoint selection. Results Totally 189 papers were included, involving 189 prescriptions of acupuncture and moxibustion, 46 acupoints, 1270 times of acupoint selection. There were 1119 times of neighboring acupoint selection (88.1%), 151 times of distant acupoint selection (11.9%), 654 times of acupoint selection in yang meridian (51.5%) and 283 times of acupoint selection in yin meridian (22.3%). Conclusion Warming acupuncture and moxibustion in clinical treatment of knee osteoarthritis focuses on acupoints in Stomach Meridian of Foot-Yangming, Spleen Meridian of Foot-Taiyin; the neighboring acupoint selection is the main method, combined with evidence-based distant acupoint selection; acupoint selection from yang meridian was emphasized.

BACKGROUND: Recent studies have demonstrated that sterol regulatory element binding protein-2 (SREBP-2) plays a key role in osteoarthritis, but its exact pathogenesis remains incompletely understood yet. OBJECTIVE:To investigate the expression of SREBP-2 in the process of interleukin-1β-induced articular chondrocyte degenerationin vitro. METHODS: Articular chondrocytes obtained from C57BL/6J mice were culturedin vitro. After the second passage, cels were randomly divided into four groups: control group, and three experimental groups treated with 10 μg/L interleukin-1β for 24, 48 and 72 hours, respectively. RESULTS AND CONCLUSION:The cels became hypertrophic after being stimulated by interleukin-1β, and the staining of colagen X was positive at 72 hours. MTT assay demonstrated that the cel activity after stimulation with interleukin-1β decreased with time. Results of RT-PCR showed that the expression of SREBP-2 and SREBP cleavage activating protein mRNA was significantly increased after stimulation with interleukin-1βas compared with the control group and increased with time. On the contrary, the expression of aggrecan and colagen II mRNA was decreased with time. It is revealed that interleukin-1β could inhibit the proliferation of regular chondrocytes and the expression of its extracelular matrix, and furthermore, induce chondrocyte hypertrophy. The expression of SREBP-2 showed a negative relationship with key cartilage genes during this interleukin-1β-induced degeneration.

Objective To compare the advantages and values of several special staining methods of chondrocytes . Methods Twelve 7-day old healthy C57BL/6J mice were killed to obtain the cartilage tissue of the knee joint in order to isolate the chondrycytes .Type II collagen was used to assess the chondrocytes .Then the chondrocyte climbing slices were prepared.The materials were fixed, and HE staining, Safranin O-fast green staining, SA-β-gal staining and immunohistochemical staining of Type II collagen were performed and compared .Results HE staining showed clear morphology of the chondrocytes .The cell nuclei were stained blue and the cytoplasm was pink .Safranin O-fast green staining showed that the nuclei were pink and the cytoplasm green .SA-β-gal staining showed that the aging cells were green while the young cells were colorless .Immunohistochemical staining of type II collagen showed the distribution of type II collagen and they were stained brown while the cell nuclei were blue .Conclusions HE staining and safranin O-fast green staining can provide more information than the other staining techniques .SA-β-gal staining is useful in the analysis of aging chondrocytes .Immunohistochemical of type II collagen can be used to study type II collagen .

Objective To develop the technique to detect total core antigen of HCV(Total HCV-cAg) by Enzyme-Linked Immu-nosorbent Assay (ELISA) and apply it for clinical diagnosis. Methods 201 serum samples with anti-HCV antibody were detected total HCV-cAg after pre - treating the samples, then the sensitivity of results were compared with HCV RNA tests. Among them, 176 cases was determined by FQ-PCR, and 25 cases by RT-PCR for HCV-RNA. Results HCV RNA was found in sera from 88 of 201 samples (43.8%). Total HCV-cAg was positive in 71 (35.3%) of 201 samples . There was no significant difference between the detection rate of HCV RNA by PCR and total HCV-cAg by ELISA. Conclusion Detection of total core antigen of HCV is suitable to be used as to diagnose HCV in clinic.