OBJECTIVE: To establish a visual reporting system for evaluating the activity of collagen Ⅰ α 1 chain (COL1A1) gene promoter in immortalized human hepatic stellate cells, so as to estimate the activation status of the cells and provide a new cell model for the screening and study of anti-hepatic fibrosis drugs. METHODS: The promoter sequence of human COL1A1 was amplified from the genomic DNA of human hepatocarcinoma cell line HepG2. Based on the pLVX-AcGFP1-N1 plasmid, the recombinant plasmid pLVX-COL1A1-enhanced green fluorescent protein (EGFP) was constructed, in which the enhanced green fluorescent protein gene expression was regulated by the COL1A1 promoter. The monoclonal cell line was acquired by stably transfecting pLVX-COL1A1-EGFP into the immortalized human hepatic stellate cell line LX-2 by the lentivirus packaging system and screening. The cell line was treated with transforming growth factor-β1 (TGF-β1) or co-treated with TGF-β1 and drugs with potential anti-hepatic fibrosis effects. The EGFP fluorescence intensity in cells was analyzed by the fluorescence microscope and ImageJ 1.49 software using a semi-quantitative method. The COL1A1 and EGFP mRNA were detected by reverse transcription real-time quantitative PCR (RT-qPCR), and corresponding proteins were detected by Western blot. RESULTS: The recombinant plasmid pLVX-COL1A1-EGFP with the expression of EGFP regulated by COL1A1 promoter was successfully constructed. Kozak sequence was added to enhance the expression of EGFP, which was identified by double digestion and sequencing. The LX-2 monoclonal cell line LX-2-CE stably transfected with pLVX-COL1A1-EGFP was obtained. After co-treatment with TGF-β1 and 5 μmol/L dihydrotanshinone Ⅰ with potential anti-hepatic fibrosis effect for 24 h, the total fluorescence intensity and the average fluorescence intensity of LX-2-CE were lower than those in TGF-β1 single treatment group (P < 0.05), the intracellular mRNA and protein levels of COL1A1 and EGFP were also lower than those in the TGF-β1 single treatment group (P < 0.05). CONCLUSION A reporter system for estimating activation of hepatic stellate cells based on COL1A1 promoter regulated EGFP expression is successfully constructed, which could visually report the changes in COL1A1 expression, one of the activation-related markers of hepatic stellate cells, in vitro. It provides a new cell model for the screening and study of anti-hepatic fibrosis drugs.
Humans ; Transforming Growth Factor beta1/pharmacology* ; Hepatic Stellate Cells/pathology* ; Liver Cirrhosis/genetics* ; Collagen Type I/pharmacology* ; RNA, Messenger/metabolism*

By referring to the relevant ancient herbal literature， medical records and prescription books， the textual research of Violae Herba has been conducted to verify the name， origin， producing area， quality evaluation and processing method changes. The results showed that the name of Zihua Diding originated from its flower color and plant morphological characteristics. The primitive plant of Violae Herba is Viola genus of Violaceae， V. yedoensis， as stipulated in the 2020 edition of Chinese Pharmacopoeia， has been the mainstream in past dynasties of China. Violae Herba is mainly wild， and it is widely distributed throughout the country. Since modern times， the quality of Violae Herba is better with integrity， green color and yellow root. There are few records on the harvesting and processing methods of Violae Herba in ancient times， most of which are directly used after drying. It is suggested that the collection and processing methods of Violae Herba in the famous classical formulas can be implemented in accordance with the provisions of the 2020 edition of Chinese Pharmacopoeia.

Background::Accurate prediction of ischemic stroke is required for deciding anticoagulation use in patients with atrial fibrillation (AF). Even though only 6% to 8% of AF patients die from stroke, about 90% are indicated for anticoagulants according to the current AF management guidelines. Therefore, we aimed to develop an accurate and easy-to-use new risk model for 1-year thromboembolic events (TEs) in Chinese AF patients.Methods::From the prospective China Atrial Fibrillation Registry cohort study, we identified 6601 AF patients who were not treated with anticoagulation or ablation at baseline. We selected the most important variables by the extreme gradient boosting (XGBoost) algorithm and developed a simplified risk model for predicting 1-year TEs. The novel risk score was internally validated using bootstrapping with 1000 replicates and compared with the CHA 2DS 2-VA score (excluding female sex from the CHA 2DS 2-VASc score). Results::Up to the follow-up of 1 year, 163 TEs (ischemic stroke or systemic embolism) occurred. Using the XGBoost algorithm, we selected the three most important variables (congestive heart failure or left ventricular dysfunction, age, and prior stroke, abbreviated as CAS model) to predict 1-year TE risk. We trained a multivariate Cox regression model and assigned point scores proportional to model coefficients. The CAS scheme classified 30.8% (2033/6601) of the patients as low risk for TE (CAS score = 0), with a corresponding 1-year TE risk of 0.81% (95% confidence interval [CI]: 0.41%-1.19%). In our cohort, the C-statistic of CAS model was 0.69 (95% CI: 0.65-0.73), higher than that of CHA 2DS 2-VA score (0.66, 95% CI: 0.62-0.70, Z = 2.01, P = 0.045). The overall net reclassification improvement from CHA 2DS 2-VA categories (low = 0/high ≥1) to CAS categories (low = 0/high ≥1) was 12.2% (95% CI: 8.7%-15.7%). Conclusion::In Chinese AF patients, a novel and simple CAS risk model better predicted 1-year TEs than the widely-used CHA 2DS 2- VA risk score and identified a large proportion of patients with low risk of TEs, which could potentially improve anticoagulation decision-making. Trial Registration::www.chictr.org.cn (Unique identifier No. ChiCTR-OCH-13003729).

OBJECTIVE: To study and compare the effect of indwelling drainage tube and extubation time on occult hemorrhage and functional recovery after total hip arthroplasty(THA). METHODS: From July 2017 to June 2018, 123 patients who underwent THA in our hospital for the first time were selected as the subjects of study. According to whether the drainage tube was retained or not and the time of extubation, they were divided into three groups:in group A, 41 patients (24 males, 17 females, age 53 to 77 years) did not put drainage tube after THA;in group B, 41 patients were removed 24 hours after THA, 26 males and 15 females, aged 55 to 74 years;in group C, 41 patients were removed 48 hours after THA, 25 males and 16 females, aged 52 to 75 years. The VAS score of pain 72 hours after THA, the total and recessive blood loss, the time of starting functional exercise, and the incidence of postoperative limb swelling were recorded. All the patients were followed up for one year after discharge. Harris hip score was used to evaluate the degree of hip function recovery one year after operation. RESULTS: The occult blood loss of group A, B and C were(513.6±25.3), (521.7±33.4), (519.3±29.8) ml, respectively, with no significant difference(>0.05). There was no significant difference in blood loss in operation among the three groups(>0.05). In group B and C, the postoperative apparent blood loss was more than that in group A (<0.05). There was no significant difference in VAS scores of the three groups before and 72 hours after operation (>0.05). The time of getting out of bed in group A was shorter than that in group B and C (<0.05), and that in group B was shorter than that in group C(<0.05). The Harris hip score at 1 year after operationof the three groups was significantly higher than that of before operation (<0.05). There was no significant difference in Harris hip score before and after operation among three groups (>0.05). There was no significant difference in the incidence of complications among three groups (>0.05). CONCLUSION Whether the drainage tube is left or not and the time of extubation have no significant effect on the latent blood loss and functional recovery after THA, but without drainage tube after THA can reduce the apparent blood loss, patients can get out of bed at 6 hours after THA, which is more conducive to the recovery and nursing of patients.
Aged ; Airway Extubation ; Arthroplasty, Replacement, Hip ; Drainage ; Female ; Humans ; Male ; Middle Aged ; Recovery of Function ; Treatment Outcome

Objective To summarize the treatment decision-making strategy and its long-term efficacy for advanced elderly patients with severe valvular heart disease and clear indications for surgery.Methods Clinical data of 196 patients aged 75 years and older firmly diagnosed as severe valvular heart diseases were retrospectively analyzed.The patients were divided into the surgical group (a mean age of 77.4±2.0 years,n=126)and the conservative group(a mean age of 80.5±5.0 years,n =70).Factors affecting therapeutic decision-making were analyzed,and the differences in a long-term survival were compared between the two groups.Results The most common reason for choosing conservative treatment was the recommendation of the doctor giving a preliminary diagnosis and worrying about the high-risk surgery for the patients(62.9％,44/70).Only 26(37.1％)patients in the conservative group were evaluated by cardiac surgeons,among whom 12 (17.1％)patients were considered to have surgical contraindications,and 14 (20.0％) patients themselves or their family members chose conservative treatment for the fear of surgical risks.Patients in the operation group were mainly from the outpatient department of cardiac surgery,and only 8 (6.3 ％)cases were referred from department of internal medicine.Logistic regression analysis showed that female,chronic renal insufficiency,advanced age,pneumonia and emergency hospital admissions were independent predictors for the conservative option(P ＜0.01),while patients with isolated aortic valve disease tended to receive surgical treatment.Overall 5-year survival was higher in the surgical group than in the conservative group (76.4％ vs.39.9％,P ＜ 0.01).Cox regression analysis disclosed that the conservative treatment option was the single risk factor for long-term survival in all series.Conclusions Many factors affect the process of therapeutic decision-making for patients with severe valvular heart diseases,and a multidisciplinary collaboration is the best way for the optimal treatment strategy for those patients.

Objective To investigate the significance of the expression of phosphatidic acid phosphatase type 2 domain containing 1A(PPAPDC1A) in human colorectal cancer cell lines.Methods The high metastatic potential cells LOVO,SW620 and low metastatic potential cells SW480,RKO,HCT116 and DLD-1 were cultured,the expression of PPAPDC1A mRNA and protein in different colorectal cancer cells in logarithmic growth period was detected by real-time quantitative polymerase chain reaction and Western blot.Results There were significant differences in the expressions of PPAPDC1A mRNA and protein among the six human colorectal cancer cells (F =41.213,344.1 16;P ＜ 0.05).The expression of PPAPDC1 A mRNA and protein in highly metastatic potential cells LOVO and SW620 was significantly higher than that in DLD-1,HCT116,RKO and SW480 cells (P ＜0.05).The expression of PPAPDC1A protein in LOVO cells with high metastatic potential was significantly higher than that in SW620 cells(P ＜ 0.05).The expression of PPAPDC1A protein in DLD-1 cells was significantly higher than that in HCT116,RKO and SW480 cells (P ＜0.05).The expression of PPAPDC1 A protein in HCT116 cells with low metastatic potential was significantly higher than that in RKO and SW480 cells (P ＜ 0.05).The expression of PPAPDC1 A protein in RKO cells was significantly higher than that in SW480 cells (P ＜ 0.05).There was no significant difference in the expression of PPAPDC1A mRNA between LOVO and SW620 cells (P ＜ 0.05).There was no significant difference in the expression of PPAPDC1A mRNA between SW480,RKO,HCT116 and DLD-1 cells (P＜ 0.05).Conclusion PPAPDC1A expresses differentially in colorectal cancer cell lines,which may be involved in the invasion and metastasis of colorectal cancer.

Objective At present, the model of pulmonary blast injury is characterized by complexity, a certain degree of safety risks,and difficulty in obtaining the materials.This article aimed to establish a simple, safe and reusable primary pulmonary blast injury model in rats. Methods A total of 16 SD rats were randomly divided into control group and pulmonary blast injury group. In rats of control group, only ketamine(120 mg/kg)was injected intraperitoneally and hair was removed from chest.In pulmonary blast injury group,after the rats were fully anesthetized and depilated on chest,they were placed in a reducing T-pipe with only chest fully exposed,and the pneumatic simulated blasting device was adjusted to 0.4 Mpa.Taking locations about 2.5 cm from the intersection points of the median line of the sternum with left and right anterior axillary lines as targets,explosion were carried out respectively,fol-lowing the sequence of right chest 2 min prior to left chest.Lung pathology,lung water content,arterial blood gas analysis, and inter-leukin-6(IL-6)changes were observed. Results No obvious dam-age was found as to the appearance of chest in all rats after blast inju-ry.Transient purpura appeared in 37.5%(3/8)of the rats within 5 min after injury and disappeared within 1 h.On the spot,62.5%(5/8)of the rats presented 10-20 s of brief apnea and gradually recov-ered 6 h later.The lung tissue W/D,serum IL-6 level and lung tissue IL-6 level in pulmonary blast injury group were significantly greater than those in control group[(4.97±0.37)vs(4.62±0.07),(68.13± 59.40)vs(16.27 ±5.08)pg/mL,(487.04 ±30.57)vs(172.74 ±35.02)pg/mL],representing statistically significant difference(P<0.05).PaO2level in pulmonary blast injury group was significantly lower than that of control group,and the difference was statistically significant(P<0.05).After pulmonary blast injury, punctate and flaky hemorrhages were observed in the lesions of lung tissues,showing red hepatic changes.Lung interstitium was obviously thickened and alveoli collapsed under microscope.Erythrocytes and edema fluid in lung interstitium and alveoli exuded.Gas embolism in intersti-tial blood vessels was observed under microscope. Conclusion The lungs of rats after blast through simple T-pipe is typical of prima-ry pulmonary blast injury in pathological and pathophysiological manifestations, which proves the convenience and reliability of the modeling method.

Objective: To verify the safety and efficacy of IONTRIS particle therapy system (IONTRIS) in clinical implementation. Methods: Between 6.2014 and 8.2014, a total of 35 patients were enrolled into this trial: 31 males and 4 females with a median age of 69 yrs (range 39-80). Ten patients had locally recurrent head and neck tumors after surgery, 4 cases with thoracic malignancies, 1 case with hepatocellular carcinoma, 1 case with retroperitoneal sarcoma, and 19 cases with non-metastatic prostate carcinomas. Phantom dose verification was mandatory for each field before the start of radiation. Results: Twenty-two patients received carbon ion and 13 had proton irradiation. With a median follow-up time of 1 year, all patients were alive. Among the 16 patients with head and neck, thoracic, and abdominal/pelvic tumors, 2, 1, 12, and 1 cases developed complete response, partial response, stable disease, or disease progression, respectively. Progression-free survival rate was 93.8% (15/16). Among the 19 patients with prostate cancer, biological-recurrence free survival was 100%. Particle therapy was well tolerated in all 35 patients. Twenty-five patients (71.4%) experienced 33 grade 1 acute adverse effects, which subsided at 1 year follow-up. Six (17.1%) patients developed grade 1 late adverse effects. No significant change in ECOG or body weight was observed. Conclusions IONTRIS is safe and effective for clinical use. However, long term follow-up is needed to observe the late toxicity and long term result.

Objective To verify the safety and efficacy of IONTRIS particle therapy system ( IONTRIS) in clinical implementation. Methods Between 6.2014 and 8.2014, a total of 35 patients were enrolled into this trial:31 males and 4 females with a median age of 69 yrs ( range 39?80) . Ten patients had locally recurrent head and neck tumors after surgery, 4 cases with thoracic malignancies, 1 case with hepatocellular carcinoma, 1 case with retroperitoneal sarcoma, and 19 cases with non?metastatic prostate carcinomas. Phantom dose verification was mandatory for each field before the start of radiation. Results Twenty?two patients received carbon ion and 13 had proton irradiation. With a median follow?up time of 1 year, all patients were alive. Among the 16 patients with head and neck, thoracic, and abdominal/pelvic tumors, 2, 1, 12, and 1 cases developed complete response, partial response, stable disease, or disease progression, respectively. Progression?free survival rate was 93.8％ (15/16). Among the 19 patients with prostate cancer, biological?recurrence free survival was 100％. Particle therapy was well tolerated in all 35 patients. Twenty?five patients (71.4％) experienced 33 grade 1 acute adverse effects, which subsided at 1 year follow?up. Six ( 17.1％) patients developed grade 1 late adverse effects. No significant change in ECOG or body weight was observed. Conclusions IONTRIS is safe and effective for clinical use. However, long term follow?up is needed to observe the late toxicity and long term result.