Objective To investigate the effect of Rho kinase inhibitor,fasudil,on pulmonary fibrosis induced by paraquat in rats in order to elucidate the underlying mechanisms.Methods A total of 72 SpragueDawley male rats of specific pathogen free (SPF) were randomly (random number) divided into four groups:the normal control group (NS group,n =18),fasudil control group (FS control group,n =18),paraquat poisoning group (PQ poisoning group,n =18) and fasudil intervention group (FS intervention group,n =18).On days 7,14,28 after paraquat exposure,six rats were respectively selected from each group.These rats were anesthetized and sacrificed immediately,and their lung tissues were collected.The hydroxyproline (HYP) in the lung tissue was detected by using alkaline hydrolysis.The expressions of type Ⅰ,Ⅲ collagen protein,connective tissue growth factor (CTGF) and ROCK1 mRNA in Rho/ROCK signaling pathway were assayed by using the real-time quantitative PCR (RT-PCR),and the levels of type Ⅰ,Ⅲ collagen protein,connective tissue growth factor (CTGF) and Rho / ROCK signaling pathway ROCK1 protein were measured by using Western blotting.The pathological changes of lung tissue were observed under light microscope.Results There were no significant differences in the observed biomarkers between FS control group and NS group (P ＞ 0.05).While in PQ poisoning group and FS intervention group on days 7,14,28 (all P ＜ 0.05),the amount of HYP increased obviously (P ＜ 0.05),the expressions of type Ⅰ,Ⅲ collagen protein,CTGF,ROCK1 mRNA and protein levels were increased significantly (P ＜ 0.05).Compared with the PQ poisoning group,the amount of HYP decreased significantly,and the expressions of type Ⅰ,Ⅲ collagen protein,CTGF,ROCK1 mRNA and protein levels were decreased significantly in FS intervention group on days 7,14,28 (all P ＜ 0.05).The pathological changes of lung tissue revealed that the degree of pulmonary fibrosis in the PQ poisoning group were most serious on 28 d after paraquat exposure,and the degree of pulmonary fibrosis were lessened in FS intervention group on days 7,14,28.Conclusions ROCK inhibitor,fasudil,has obvious therapeutic effects on paraquat-induced lung fibrosis,by regulating Rho / ROCK signaling pathway with downregulated expression of CTGF,and decrease in the levels of type Ⅰ,Ⅲ collagen protein,thus reducing protein deposition.

OBJECTIVE：To establish characteristic chromatogram of Cornus officinalis and its different wine-processedproducts，investigate the differences of chromaticity values，and analyze them with chemical pattern recognition technology.METHODS：UPLC method was adopted. Using loganin as reference，UPLC characteristic chromatograms were drawn for 10batches of C. officinalis and 20 batches of different wine-processed products （stewing with wine，steaming with wine）. TCMFingerprint Similarity Evaluation System（2012A edition）was used for similarity evaluation，and common peaks were confirmed.The chromaticity values [lightness（L），red and green tone value（a），yellow and blue tone value（b），color difference value（ΔE）]were determined by spectrophotometer. SPSS 20.0 and SIMCA 14.0 software were used for cluster analysis，principal componentanalysis and partial least squares-discriminant analysis；taking the area of characteristic peak and chromaticity value as indexes，andthe variable importance projection greater than 1 as the standard，the difference markers affecting its quality were screened.RESULTS：There were 6 common peaks in the chromatograms for decoction piece of C. officinalis，7 common peaks forwine-processed C. officinalis（stewing with wine）and wine-processed C. officinalis（steaming with wine）. Four components wereidentified as gallic acid，5-hydroxymethylfurfural，morroniside，loganin. 5-hydroxymethylfurfural was produced after processing.The similarity between C. officinalis and different wine-processed products （stewing and steaming with wine） was low（0.869-0.937，0.845-0.944），but the similarity between different wine-processed products was higher than 0.99. ΔL，Δa，Δb and ΔEof C. officinalis decoction pieces and wine-processed C. officinalis decoction pieces（stewing in wine）were －9.42-－3.58，－24.92- －15.00，－11.33- －7.00 and 17.01-28.12，respectively. ΔL，Δa，Δb and ΔE of C. officinalis decoction pieces and wine-processed C. officinalis（steaming in wine）decoction pieces were －8.58-－2.42，－25.08-－13.83，－10.92-－6.08，15.58-28.67. ΔL，Δa，Δb and ΔE of wine-processed C. officinalis decoction pieces（stewing and steaming with wine）were －2.17-3.00，－0.75-2.50， 0.25-1.42 and 1.25-3.83，respectively. Results of cluster analysis showed that 30 batches of sample were clustered into two categories，S1-S10 were clustered into one category，and S11-S30 were clustered into other category. Principal component analysis showed that cumulative contribution rate of former two main components was 83.147％. Results of partial least squares-discriminant analysis showed that morroniside，No.5 peak and chromaticity values（L，a，b）were the difference markers affecting its quality. CONCLUSIONS：Established UPLC characteristic chromatogram is stable and feasible，and can be used to rapidly identify C. officinalis and its different wine-processed products. Established chemical mode can be used to identify different wine-processed products.

Objective To explore the role of CD4~+CD25~+CD127~(lo) regulatory T cells (Tregs) and inter-leukin (IL)-6, transforming growth factor beta (TGF-β), IL-17 in the pathogenesis of lupus nephritis (LN) by detecting the levels of IL-10, IL-6, TGF-β, IL-17, CD4~+CD25~+CD127~(lo) Tregs in the peripheral blood of patients with active and inactive LN. Methods Three-colour flow cytometry was used to quantitatively measure proportions of Treg cells, the levels of TGF-β, IL-17 were detected by ELISA, and the levels of IL-10, IL-6 in the peripheral blood were detected by Cytometric Bead Array System. Results ① Compared with the inactive LN and the normal controls (P<0.01), the level of CD4~+CD25~+CD127~(lo) Tregs from patients with active LN was lower(P<0.01). When compared with the normal controls, the level of CD4~+CD25~+CD127~(lo) Tregs from LN inactive patients had no significant difference (P>0.05). ② Compared with patients with inactive LN, the levels of IL-10, IL-6 was higher (P<0.01) in patients with active LN. ③ Compared with the patients with inactive LN and the normal controls, the levels of TGF-β, IL-17 was not significantly different (P>0.05). ④ The level of CD4~+CD25~+CD127~(lo) T cell was correlated negatively with the levels of IL-10, IL-6 and SLEDAI (P<0.05), and was not correlated with C3 and C4. ⑤ SLEDAI was correlated positively with the levels of IL-10 and IL-6 (P<0.01). SLEDAI and the level of IL-10 were correlated negatively with C3 and C4 (P<0.01 for both). ⑥ The level of CD4~+CD25~+CD127~(lo) Tregs from LN was not correlated with TGF-β and IL-17. ⑦ TGF-β was correlated positively with the level of IL-17. Conclusion ① The level changes of Tregs and IL-10, IL-6, TGF-β in the peripheral blood of LN can be used as the indicators for the activity status of lupus nephritis. ② Tregs and IL-10, IL-6 in the peripheral blood of LN patients is negatively correlated. ③ The glucocorticoid hormone is helpful to elevate the level of Tregs but decrease IL-17. T cell level can vary in different body status, different microenvironmental and immune status.

Objective: To explore the effect of interfering insulin-like growth factors-1 receptors (IGF-1R) by small interfering RNA (siRNA) on cell cycle and apoptosis of hypoxic hepatocellular carcinoma HepG2 cells. Methods: The hypoxic hepatocellular carcinoma model was established via cobalt chloride treatment. Three siRNAs targeting IGF1R gene and one negative control siRNA were designed and synthesized. They were transfected into hypoxic HepG2 cells, and 24 h later, the transfection efficiency was detected by fluorescent microscopy. The protein expression of IFG-1R was detected with Western blotting (WB) to screen the siRNA with highest transfection efficacy. The selected siRNA was used to transfect hypoxic HepG2 cells. The proliferation of hypoxic HepG2 cells was determined by MTT assay. Cell cycle distribution and apoptosis were analyzed by Flow cytometry. WB was performed to detect the proteinexpressionsofCDK1,CDK2andCaspase-3inHepG2cells. Results: The hypoxic hepatocellular carcinoma model was successfully established. IGF-1R-siRNA-2 showed the most effective interference efficiency and the most significant knockdown of IGF-1R (all P<0.01). The proliferation of HepG2 cells transfected with IGF-1R siRNA-2 was significantly suppressed (P<0.05 or P<0.01), the cell cycle was blocked at G0/G1 phase (P<0.05), and the apoptosis rate was increased up to (25.3±1.3)% P<0.01). In the meanwhile, the expressions of CDK1 and CDK2 were decreased and the expression of Caspase-3 was increased in hypoxic HepG2 cells after IGF-1R knockdown (P<0.05). Conclusion: Interfering IGF-1R by siRNA inhibits the malignant biological behaviors of hypoxic HepG2 cells via regulating cell cycle and apoptosis-related proteins. IGF-1R may be a potential target for the treatment of HCC.

Objective:To analyze the correlation of fetal cervical cystic hygroma (CCH) with chromosomal and structural abnormalities and to assess the prognosis of CCH.Methods:This study retrospectively enrolled 70 fetuses with CCH diagnosed by prenatal ultrasound in the First Affiliated Hospital of Xi'an Jiao Tong University from July 2015, to December 2021. According to whether complicated by structural malformations or other anomalies, all the subjects were divided into the non-isolated and isolated CCH groups. The correlation of CCH and the gestational age at detection with chromosomal and structural abnormalities were analyzed and the prognosis of the cases were summarized using Chi-square test. Results:There were 34 isolated CCH (34/70, 49%) and 36 non-isolated CCH (36/70, 51%) among the 70 cases. In the non-isolated CCH group, there were eight cases (22%, 8/36) with abnormal heart structure, ten (28%, 10/36) with abnormal anterior abdominal wall, 16 (44%,16/36) with systemic edema and/or pleural effusion, one (3%,1/36) with craniocerebral abnormalities and one with holoprosencephaly and cardiac structural abnormalities. Eighteen out of 44 cases undergoing chromosome testing had chromosomal abnormalities, which were trisomy-18 ( n=6), trisomy-21 ( n=3), trisomy-13 ( n=3), 45,XO ( n=3), and chromosome segment duplication or deletion ( n=3). The detection rate of chromosome abnormality was higher in non-isolated CCH group comparing with isolated CCH group [59%(13/22) vs 23%(5/22), χ2=6.02, P=0.014]. There was no significant difference in the gestational age at the detection of CCH or proportion of women of advanced maternal age between the isolated and non-isolated CCH groups (both P>0.05). The ratios of isolated CCH cases with normal chromosome detected at the gestational weeks of 14-27 +6 was higher than those detected at 11-13 +6 weeks [62%(13/21) vs 17%(4/23), χ2=7.39, P=0.001]. Out of the 17 cases with isolated CCH and normal chromosomes, 12 were live births. One of the 12 cases still had a cystic mass with a diameter of 3 cm in the neck nine months after birth, and the other 11 cases had no mass at birth but one case died at the age of five months (hospitalized one week for neonatal edema),one case was found with anal atresia three days after birth and underwent operation and the remaining nine cases were normal during five months to six years follow-up. Conclusions:Non-isolated CCH is at a higher risk of chromosomal abnormalities. Isolated CCH cases detected later had higher rate of normal chromosome and often have a higher survival rate.