Clinical value of plasma scaffold protein SEC16A in evaluating hepatitis B-related liver cirrhosis and hepatocellular carcinoma / 中华肝脏病杂志

Objective: To investigate the clinical value of plasma scaffold protein SEC16A level and related models in the diagnosis of hepatitis B virus-related liver cirrhosis (HBV-LC) and hepatocellular carcinoma (HBV-HCC). Methods: Patients with HBV-LC and HBV-HCC and a healthy control group diagnosed by clinical, laboratory examination, imaging, and liver histopathology at the Third Hospital of Hebei Medical University between June 2017 and October 2021 were selected. Plasma SEC16A level was detected using an enzyme-linked immunosorbent assay (ELISA). Serum alpha-fetoprotein (AFP) was detected using an electrochemiluminescence instrument. SPSS 26.0 and MedCalc 15.0 statistical software were used to analyze the relationship between plasma SEC16A levels and the occurrence and development of liver cirrhosis and liver cancer. A sequential logistic regression model was used to analyze relevant factors. SEC16A was established through a joint diagnostic model. Receiver operating characteristic curve was used to evaluate the clinical efficacy of the model for liver cirrhosis and hepatocellular carcinoma diagnosis. Pearson correlation analysis was used to identify the influencing factors of novel diagnostic biomarkers. Results: A total of 60 cases of healthy controls, 60 cases of HBV-LC, and 52 cases of HBV-HCC were included. The average levels of plasma SEC16A were (7.41 ± 1.66) ng/ml, (10.26 ± 1.86) ng/ml, (12.79 ± 1.49) ng /ml, respectively, with P < 0.001. The sensitivity and specificity of SEC16A in the diagnosis of liver cirrhosis and hepatocellular carcinoma were 69.44% and 71.05%, and 89.36% and 88.89%, respectively. SEC16A, age, and AFP were independent risk factors for the occurrence of HBV-LC and HCC. SAA diagnostic cut-off values, sensitivity, and specificity were 26.21 and 31.46, 77.78% and 81.58%, and 87.23% and 97.22%, respectively. The sensitivity and specificity for HBV-HCC early diagnosis were 80.95% and 97.22%, respectively. Pearson correlation analysis showed that AFP level was positively correlated with alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil), and γ-glutamyltransferase (GGT) with P < 0.01, while the serum SEC16A level was only slightly positively correlated with ALT and AST in the liver cirrhosis group (r = 0.268 and 0.260, respectively, P < 0.05). Conclusion: Plasma SEC16A can be used as a diagnostic marker for hepatitis B-related liver cirrhosis and hepatocellular carcinoma. SEC16A, combined with age and the AFP diagnostic model with SAA, can significantly improve the rate of HBV-LC and HBV-HCC early diagnosis. Additionally, its application is helpful for the diagnosis and differential diagnosis of the progression of HBV-related diseases.

Study on HBV-related acute-on-chronic liver failure risk factors and novel predictive survival model / 中华肝脏病杂志

Objective: To identify the predisposing factors, clinical characteristics, and risk factors of disease progression to establish a novel predictive survival model and evaluate its application value for hepatitis B virus-related acute-on-chronic liver failure. Methods: 153 cases of HBV-ACLF were selected according to the guidelines for the diagnosis and treatment of liver failure (2018 edition) of the Chinese Medical Association Hepatology Branch. Predisposing factors, the basic liver disease stage, therapeutic drugs, clinical characteristics, and factors affecting survival status were analyzed. Cox proportional hazards regression analysis was used to screen prognostic factors and establish a novel predictive survival model. The receiver operating characteristic curve (ROC) was used to evaluate predictive value with the Model for End-Stage Liver Disease (MELD) and the Chronic Liver Failure Consortium Acute-on-Chronic Liver Failure score (CLIF-C ACLF). Results: 80.39% (123/153) based on hepatitis B cirrhosis had developed ACLF. HBV-ACLF's main inducing factors were the discontinuation of nucleos(t)ide analogues (NAs) and the application of hepatotoxic drugs, including Chinese patent medicine/Chinese herbal medicine, non-steroidal anti-inflammatory drugs, anti-tuberculosis drugs, central nervous system drugs, anti-tumor drugs, etc. 34.64% of cases had an unknown inducement. The most common clinical symptoms at onset were progressive jaundice, poor appetite, and fatigue. The short-term mortality rate was significantly higher in patients complicated with hepatic encephalopathy, upper gastrointestinal hemorrhage, hepatorenal syndrome, and infection (P < 0.05). Lactate dehydrogenase, albumin, the international normalized ratio, the neutrophil-to-lymphocyte ratio, hepatic encephalopathy, and upper gastrointestinal bleeding were the independent predictors for the survival status of patients. The LAINeu model was established. The area under the curve for evaluating the survival of HBV-ACLF was 0.886, which was significantly higher than the MELD and CLIF-C ACLF scores (P < 0.05), and the prognosis was worse when the LAINeu score ≥ -3.75. Conclusion: Discontinuation of NAs and the application of hepatotoxic drugs are common predisposing factors for HBV-ACLF. Hepatic decompensation-related complications and infection accelerate the disease's progression. The LAINeu model can predict patient survival conditions more accurately.

Effects of Tuina on Muscle-specific microRNA and Factors Related with Proliferation and Differentiation of Muscle Satellite Cells in Denervation-induced Atrophy in Rats / 中国康复理论与实践

Objective:To investigate the effects and mechanism of Tuina on denervation-induced atrophy. Methods:A total of 42 Sprague-Dawley rats were randomly divided into sham group (n = 6), model group (n = 18) and Tuina group (n = 18). The model group and Tuina group freed and excised right tibia nerve about one centimeter, while the sham group freed the right tibia nerve only. From the second day after operation, Tuina group accepted Tuina on the injured area, while the sham group and the model group were only fixed without any intervention. Six rats were sacrificed on the 14th, 21st and 28th day after operation in the model and Tuina groups, and the sham group was sacrificed on the 28th day after operation. The gastrocnemius muscles were measured wet weight ratio. The diameter and area of muscle cells were measured under HE staining. The expression of Pax7, MyoD, MyoG, microRNA-1, microRNA-133a and microRNA-206 in the gastrocnemius muscles were detected with reverse transcription real-time quantitative polymerase chain reaction. Results:Compared with the sham group, the wet weight ratio, the area of muscle cells (except the 14-day-Tuina group) and the diameter of muscle cells decreased at each time point in the model group and Tuina group (P < 0.05); compared with the model group, the wet weight ratio, muscle cell diameter and muscle cell area increased at each time point in Tuina group (P < 0.05). Compared with the sham group, the expression of Pax7 increased in the 14-day-model group (P < 0.05) and decreased in the 28-day-model group (P < 0.05), and it increased at each time point (except 28-day) in Tuina group (P < 0.05); compared with the model group, the expression of Pax7 increased at each time point in Tuina group (P < 0.05). Compared with the sham group, the expression of MyoD and MyoG increased at each time point in the model group and Tuina group (P < 0.05); compared with the model group, the expression of MyoD and MyoG increased at each time point (except 14-day) in Tuina group (P < 0.05). Compared with the sham group, the expression of microRNA-1 and microRNA-133a decreased, and microRNA-206 increased in the model group and Tuina group at 21-day (P < 0.05); compared with the model group, the expression of microRNA-1, microRNA-133a and microRNA-206 increased in Tuina group (P < 0.05). Conclusion:Tuina may activate the Pax7/MyoD/MyoG pathway by increasing the expression of muscle-specific microRNA, to promote the proliferation and differentiation of muscle satellite cells, and delay denervation-induced atrophy.

Effect of Tuina on Autophagy-related Factor Beclin-1, Vacuolar Protein Sorting and Microtubule-associated Protein Light Chain 3 in Rats with Denervated Skeletal Muscle Atrophy / 中国康复理论与实践

Objective:To explore the effect and mechanism of Tuina on denervated skeletal muscle atrophy. Methods:A total of 77 male Sprague-Dawley rats were randomly divided into sham group (n = 7), model group (n = 35) and Tuina group (n = 35). The latter two groups were established skeletal muscle atrophy model by exposing and cutting off the common tibial nerve of rats. One day after modeling, the lower limbs of the surgical side received Tuina in Tuina group. Separately, the surgical side of gastrocnemius muscle were sampled on the 0th, 7th, 14th, 21st and 28thday after modeling, and measured the wet mass ratio. The cross-sectional area and diameter of muscle fiber were measured after HE staining. The mRNA expression of autophagy-realated factor Beclin-1, vacuolar protein sorting (Vps34) and microtubule-associated protein light chain 3 (LC3) were tested with reverse transcription real-time quantitative polymerase chain reaction. Results:There was no statistical difference in the ratio of gastrocnemius wet weight, the cross-sectional area and diameter of muscle fiber, and the mRNA expression of Beclin-1, Vps34 and LC3 among three groups on the 0th day (F < 1.321, P > 0.05). Compared with the sham group, the ratio of gastrocnemius wet weight, the cross-sectional area and diameter of muscle fiber decreased at different time points in the model group and Tuina group (P < 0.05), the ratio of gastrocnemius wet weight was higher, and the cross-sectional area and diameter of muscle fiber were bigger, both except on the 21st day, in Tuina group than in the model group (P < 0.05). Compared with the sham group, the mRNA expression of Beclin-1, Vps34 and LC3 increased at different points in the model group than in Tuina group (P < 0.05), and all the mRNA expression was higher, except on the 14th day, in Tuina group than in the model group (P < 0.05). The ratio of gastrocnemius wet weight, the cross-sectional area and diameter of muscle fiber showed a trend of progressive decrease with time in the model group and Tuina group (P < 0.05). The mRNA expression of Beclin-1 and Vps34 increased (P < 0.05), and the mRNA expression of LC3 increased in the model group 21 days after intervention (P < 0.05). The mRNA expression of Beclin-1, Vps34 and LC3 increased first and then decreased, except the mRNA expression on the 14th day in Tuina group (P < 0.05). Conclusion:Tuina may promote the activation of autophagy by up-regulating the expression of autophagy-realated factor Beclin-1, Vps34 and LC3, remove the damaged organelles and proteins, provide certain synthetic substrate and energy for muscle fiber regeneration, thereby reduce the loss of degree of denervated skeletal muscle atrophy.

Activated mTOR signaling pathway in myofibers with inherited metabolic defect might be an evidence for mTOR inhibition therapies / 中华医学杂志(英文版)

BACKGROUND@#Abnormally activated mechanistic target of rapamycin (mTOR) pathway has been reported in several model animals with inherited metabolic myopathies (IMMs). However, the profiles of mTOR pathway in skeletal muscles from patients are still unknown. This study aimed to analyze the activity of mTOR pathway in IMMs muscles.@*METHODS@#We collected muscle samples from 25 patients with mitochondrial myopathy (MM), lipid storage disease (LSD) or Pompe disease (PD). To evaluate the activity of mTOR pathway in muscle specimens, phosphorylation of S6 ribosomal protein (p-S6) and p70S6 kinase (p-p70S6K) were analyzed by Western blotting and immunohistochemistry.@*RESULTS@#Western blotting results showed that p-p70S6K/p70S6K in muscles from LSD and MM was up-regulated when compared with normal controls (NC) (NC vs. LSD, U = 2.000, P = 0.024; NC vs. MM: U = 6.000, P = 0.043). Likewise, p-S6/S6 was also up-regulated in muscles from all three subgroups of IMMs (NC vs. LSD, U = 0.000, P = 0.006; NC vs. PD, U = 0.000, P = 0.006; NC vs. MM, U = 1.000, P = 0.007). Immunohistochemical study revealed that p-S6 was mainly expressed in fibers with metabolic defect. In MM muscles, most p-S6 positive fibers showed cytochrome C oxidase (COX) deficiency (U = 5.000, P = 0.001). In LSD and PD muscles, p-S6 was mainly overexpressed in fibers with intramuscular vacuoles containing lipid droplets (U = 0.000, P = 0.002) or basophilic materials (U = 0.000, P = 0.002).@*CONCLUSION@#The mTOR pathway might be activated in myofibers with various metabolic defects, which might provide evidence for mTOR inhibition therapy in human IMMs.

Berberine Induces Cell Apoptosis through Cytochrome C/Apoptotic Protease-Activating Factor 1/Caspase-3 and Apoptosis Inducing Factor Pathway in Mouse Insulinoma Cells / 中国结合医学杂志

OBJECTIVE@#To investigate apoptotic effects of berberine, a significant alkaloids component existing in Rhizoma coptidis, and its possible acting mechanism in insulinoma cells.@*METHODS@#Different concentrations of berberine were used to treat mouse insulinoma (MIN6) cells for various period of time. The viability and apoptosis of the cells were analyzed using methylthiazolyldiphenvl-tetrazolium bromide assay, flow cytometry and enzyme-linked immuno sorbent assay. Changes in the relating pro- and anti-apoptosis proteins were detected by western-blotting.@*RESULTS@#The half-maximal inhibitory concentration (IC) of berberine was 5.7 μmol/L on MIN6 cells viability for 16 h. Berberine caused a 20% reduction (P<0.05) in cell number after only 4-h incubation; which reached 50% after 24 h (P<0.01). Berberine treatment for 16 h significantly increased the level of DNA fragmentation. The flow cytometry showed the apoptotic rate increased 2.9- and 4.6-fold after treating with berberine (5 μmol/L) for 8 and 16 h, while 3- and 8.7-fold after 10 μmol/L treatment for 8 and 16 h (P<0.01). Berberine treatment dramatically elevated the expression ratio of Bax to Bcl-2. Meanwhile, berberine notably increased the apoptosis-inducing factors and cytochrome C transforming from the mitochondria to the cytoplasm. Apoptotic protease-activating factor 1 (Apaf-1) was subsequently activated after cytochrome C release. Furthermore, caspase-3 and poly adenosine diphosphate-ribose polymerase were also activated to trigger apoptosis cascade.@*CONCLUSION@#High concentration (5 and 10 μmol/L) of berberine could induce the apoptosis of MIN6 cells through cytochrome C/Apaf-1/caspase-3 and apoptosis inducing factor (AIF) pathway.

Therapeutic effect of massage on denervated skeletal muscle atrophy in rats and its mechanism / 中国应用生理学杂志

OBJECTIVE@#To investigate the therapeutic effects of massage on denervated skeletal muscle atrophy in rats and its mechanism.@*METHODS@#Forty-eight male SD rats were randomly divided into model group (n=24) and massage group (n=24). Gastrocnemius muscle atrophy model was established by transecting the right tibial nerve of rat. On the second day after operation, the gastrocnemius muscle of the rats in the massage group was given manual intervention and the model group was not intervened. Six rats were sacrificed at the four time points of 0 d, 7 d, 14 d and 21 d. The gastrocnemius of the rats were obtained and measured the wet mass ratio after weighing. Cross-sectional area and diameter of the muscle fiber were measured after HE staining. The relative expressions of miR-23a, Akt, MuRF1 and MAFbx mRNA were tested with qPCR.@*RESULTS@#Compared with 0 d, the wet weight ratio, cross-sectional area and diameter of gastrocnemius muscle showed a progressive decline in the model group and massage group. The wet weight ratio, cross-sectional area and diameter of gastrocnemius muscle in the massage group were higher than those in the model group on 7 d, 14 d and 21 d (P＜0.05, P＜0.01). Compared with 0 d, the expressions of MuRF1, MAFbx and Akt mRNA were increased first and then were decreased in the model group and massage group. The expression of MuRF1 mRNA in massage group was lower than that in model group on 7 d and 21 d (P＜0.05, P＜0.01). The expression of MAFbx mRNA in massage group was lower than that in model group on 7 d, 14 d and 21 d (P＜0.01, P＜0.05, P＜0.01). The expression of Akt mRNA in massage group was higher than that in model group on 7 d, 14 d and 21 d (P＜0.05, P＜0.01). Compared with 0 d, the expression of miR-23a mRNA was increased in the model group and massage group on 21 d, and the expression of miR-23a mRNA in massage group was higher than that in model group (P＜ 0.05).@*CONCLUSION@#Massage can delay the atrophy of denervated skeletal muscle. The mechanism may be related to up-regulation of the expression of miR-23a and Akt mRNA, down-regulation of the expressions of MuRF1 and MAFbx mRNA, inhibition of protein degradation rate, and reduction of skeletal muscle protein degradation.

Changes in expression of autophagy-related factors during acute contusion repair of skeletal muscle / 中国应用生理学杂志

<p><b>OBJECTIVES</b>To explore the possible biological mechanism of skeletal muscle contusion repair through researching the changes in expression of autophagy-related genes and proteins in SD rats with acute skeletal muscle contusion.</p><p><b>METHODS</b>Six rats were randomly selected as the control group from 30 male SD rats, acute skeletal muscle contusion model were established in the remaining 24 rats with self-made hitter, then the model rats were randomly divided into 4 groups (3 d, 5 d, 7 d, 14 d groups, =6). On the 3, 5, 7 and 14 day after injury, injured gastrocnemius of each group was harvested. The morphological and the ultra-microstructure changes of gastrocnemius after injury were observed by HE staining and transmission electron microscope (TEM) respectively. The relative protein levels of (LC3-Ⅱ) and P62 of each group were observed by Western blot. The relative mRNA levels of atg7, atg10, atg12, atg16L1 of each group were observed by RTPCR.</p><p><b>RESULTS</b>The results of HE staining showed that compared with the control group, the inflammation reached its peak on the 5 day after injury, new muscle fibers were clearly observed in 7 d group. The results of TEM showed that, compared with the control group, oncotic mitochondria could be clearly seen in the 3 d, 5 d, 7 d groups. Also, the Z line changed from disappearing to drift thickening, sarcoplasmic reticulum dilatation gradually improved, there was no evident difference between the 14 d group and the control group, suggesting that the damage has preliminarily healed. The results of Western blot showed that the expressions of LC3-Ⅱand P62 were increased at first and then decreased. The expression of LC3-Ⅱwas markedly up-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (<0.01). Similarly, compared with the control group, the expression of P62 reached its peak on the 3 day after injury (<0. 01), and returned to normal level on the 14 day. The results of RT-PCR showed that the expression of atg10 mRNA in the natural recovery group of 3 d, 5 d, 7 d, 14 d was firstly decreased and then increased, the atg10 mRNA was markedly down-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (<0. 01). The expression of atg7, atg12, atg16L1 mRNA was generally increased at first and then decreased, it was markedly up-regulated in the 3 d, 5 d, 7 d groups compared with the control group and the 14 d group (<0.01, <0.05, <0.01).</p><p><b>CONCLUSIONS</b>The above results indicate that the autophagy is involved in repair of skeletal muscle injury by its autophagyrelated factors,regularly changes after contusion, and the rate of damage repair may be related to the level of autophagy.</p>

Effect of triglyceride level on acute kidney injury complicating acute pancreatitis / 上海预防医学

[Objective] To explore whether the triglyceride level is associated with incidence of acute kidney injury in patients with acute pancreatitis. [Methods] From Jan 2015 to March 2017, 184 patients with acute pancreatitis were selected and divided in to 3 groups based on different triglyceride levels: ideal triglyceride group (n = 89) , mild high triglyceride group (n = 53) and severe high triglyceride group (n =42). The incidence of acute kidney injury and its severity were compared between the three groups.[Results]The incidence of acute kidney injury in severe high triglyceride group was 33.3％, significantly higher than that in ideal triglyceride group (12.3％). The surgical treatment (16.7％ vs 4.5％) , average hospitalization days (20 days vs 14 days) and systemic inflammatory response syndrome score (3.0 vs2.3) in severe high triglyceride group were higher than those in ideal triglyceride group, and all differences between the two groups were significant. After adjusting for factors such as age, sex, body mass index and other confounders, the risk of acute kidney injury occurring in severe high triglyceride group was 2.35 times that in ideal triglyceride group (95％CI: 1.32-4.29). [Conclusion] High triglyceride level proves to be associated with high risk of acute kidney injury in patients with acute pancreatitis.

Effects of Electroacupuncture on Autophagy-related Gene Expression in Skeletal Muscle in Denervated Rats / 中国康复理论与实践

@#Objective To explore the effects and mechanism of electroacupuncture(EA)on denervation-induced atrophy. Methods A total of 21 male Sprague-Dawley rats were divided into sham group(n=7),model group(n=7)and EA group (n=7).The latter two groups were cut off their right sciatic nerve.Since one day after modeling,EA group accept-ed electroacupuncture at right Zusanli(ST36)and Huantiao(GB30)for eight weeks.Then,the gastrocnemius of all the rats were obtained,and measured the wet mass ratio.Cross-sectional area(CSA)and fiber diameter were measured after HE staining. The expression of autophagy-related gene ULK1, Atg13, Beclin1, Atg14, Atg7, Atg12,Atg5 and Atg16L1 were tested with reverse transcription real-time quantitative polymerase chain reaction. Results Compared with the sham group,the wet mass ratio,CSA and fiber diameter of gastrocnemius were lower signifi-cantly in the model group and EA group(P<0.001),and they were more in EA group than in the model group(P<0.05).Compared with the sham group,the mRNA expression of ULK1,Atg13,Beclin1,Atg14,Atg7,Atg12,Atg5 and Atg16L1 was more significantly in the model group (P<0.001), and they decreased in EA group compared with those of the model group(P<0.05). Conclusion Electroacupuncture can inhibit the overexpression of autophagy-related gene in denervated rats,which may steady skeletal muscle cells to delay atrophy.

Advance in Model Establishment for Denervated Skeletal Muscle Atrophy (review) / 中国康复理论与实践

The skeletal muscle atrophy could be induced by the injury of nerve. According to the source of denervated skeletal mus-cle atrophy, it could be divided into exogenous muscle atrophy and endogenous muscle atrophy. In recent years, the ex-ogenous muscle atrophy models are mainly established by operating, physically injuring or chemically injuring, while the endogenous muscle atrophy models are mainly established by the transgenic animals of amyotrophic lateral sclero-sis. The selection and optimazation of animal models are crucial for the basic studies of denervated skeletal muscle atro-phy.

Effects of PHPO on apoptosis and proliferation of lung cancer cell line A549 / 上海交通大学学报（医学版）

Objective · To investigate the effect of a new type of allene compound, 1-phenylpropadienyl phosphine oxide (PHPO), on proliferation and apoptosis of lung cancer cell line A549. Methods · A549 cells were treated with different concentrations of PHPO. The effects of PHPO on cell proliferation, apoptosis and cell cycle were detected by CCK-8 and flow cytometry assay. Wound healing test was used to measure the migration ability of A549 cells. Real-time PCR was used to detect the expression of apoptosis and cell cycle related gene. The expression of proteins in MAPK pathway was assayed by the Western blotting. The nude mice xenograft model of human lung cancer A549 cells was established. After tumor formation, PHPO was injected daily for treatment, and the tumor size was observed. Results · Compared to the control group, PHPO significantly inhibited the cell viability of A549 cells and induced apoptosis of them, and the IC50 value of 24 h is 44.23 μmol/L. PHPO blocked the cell cycle in the G1 phase significantly. The migration capacity of PHPO-treated cells was decreased. The mRNA levels of Bax and P21 were up-regulated in PHPO-treated group, and the mRNA lever of Bcl-2 was down-regulated (P<0.05). PHPO increased the phosphorylation levels of p38, ERK and JNK. Injection of PHPO could significantly inhibit the growth of tumor in the xenograft model compared to the control group (P<0.05). Conclusion · PHPO can induce the apoptosis and inhibit the proliferation of A549 cells, block the cell cycle in the G1 phase and decrease the migration ability of A549 cells significantly. The mechanism may be related to the activation of MAPK signaling pathway by PHPO and the increase of phosphorylation of p38, ERK and JNK.

Effects of PHPO on apoptosis and proliferation of lung cancer cell line A549 / 上海交通大学学报（医学版）

Objective · To investigate the effect of a new type of allene compound, 1-phenylpropadienyl phosphine oxide (PHPO), on proliferation and apoptosis of lung cancer cell line A549. Methods · A549 cells were treated with different concentrations of PHPO. The effects of PHPO on cell proliferation, apoptosis and cell cycle were detected by CCK-8 and flow cytometry assay. Wound healing test was used to measure the migration ability of A549 cells. Real-time PCR was used to detect the expression of apoptosis and cell cycle related gene. The expression of proteins in MAPK pathway was assayed by the Western blotting. The nude mice xenograft model of human lung cancer A549 cells was established. After tumor formation, PHPO was injected daily for treatment, and the tumor size was observed. Results · Compared to the control group, PHPO significantly inhibited the cell viability of A549 cells and induced apoptosis of them, and the IC50 value of 24 h is 44.23 μmol/L. PHPO blocked the cell cycle in the G1 phase significantly. The migration capacity of PHPO-treated cells was decreased. The mRNA levels of Bax and P21 were up-regulated in PHPO-treated group, and the mRNA lever of Bcl-2 was down-regulated (P<0.05). PHPO increased the phosphorylation levels of p38, ERK and JNK. Injection of PHPO could significantly inhibit the growth of tumor in the xenograft model compared to the control group (P<0.05). Conclusion · PHPO can induce the apoptosis and inhibit the proliferation of A549 cells, block the cell cycle in the G1 phase and decrease the migration ability of A549 cells significantly. The mechanism may be related to the activation of MAPK signaling pathway by PHPO and the increase of phosphorylation of p38, ERK and JNK.

Short term effects of 1g/L bromfenac sodium eye drops after LASEK / 国际眼科杂志(Guoji Yanke Zazhi)

?AIM: To compare postoperative anti -inflammation effect and safety between bromfenac sodium eye drops and pranoprofen eye drops in patients after laser epithelial keratomileusis ( LASEK) .? METHODS: In the prospective, randomized and controlled study, 100 patients ( 200 eyes ) undergoing LASEK were randomized into the bromfenac sodium group (100 eyes) and control group (100 eyes).Patients in bromfenac sodium group received bromfenac sodium hydrate ophthalmic solution eye drops twice a day in 3d before surgery and 2wk after surgery, while the patients from the control group were given proanoprofen eye drops 4 times a day in the same period.At 1, 3, 5d, 1 and 3mo after surgery, irritative symptoms grade, duration of irritation, time for corneal epithelial healing, cornel haze, uncorrected visual acuity and intraocular pressure ( IOP ) were observed and compared between the two groups. Quantitative data were analyzed using independent samples t-test and ranked data were statistically analyzed using the Mann-Whiteney rank sun test.?RESULTS:There was no significant difference between two groups in irritative symptoms grade ( P =0.317 ), neither was existed between two groups in uncorrected visual acuity after surgery (P>0.05).There was no statistical significance in the time for corneal epithelial healing between two groups (P=0.551).?CONCLUSION: Bromfenac sodium eye drops ( 1g/L ) can achieve the same therapeutic effect as pranoprofen eye drops after LASEK.

Analysis of the direction and degree of static cyclotorsion component and dynamic cyclotorsion component in corneal refractive surgery / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To analyze the direction and degree of static cyclotorsion component (SCC) and dynamic cyclotorsion component (DCC) in corneal refractive surgery. METHODS:Retrospective analysis. Totally 130 patients (260 eyes) with corneal refractive surgery in our hospital, according to the operation method were divided into femtosecond laser - assisted laser in situ keratomileusis (FS-LASIK) group and T-photorefractive keratectomy (T-PRK) group, the differences of the parameters of the two groups were compared; the differences of SCC success rate, SCC, DCC, and the eyeball rotation direction were compared between the two groups; correlation analysis on SCC, DCC and the parameters of postoperative patients were performed. RESULTS: High order aberrations and spherical aberration in the T-PRK group after operation was higher than those of FS - LASIK group, and the difference was statistically significant (P0. 05); DCC in T-PRK group (2. 86o±1. 14o) was higher than that of FS-LASIK group ( 2. 17o ± 1. 09o), and the difference was statistically significant (P0. 05). The SCC of subjects in operation was positively correlated with UCVA, BCVA, spherical equivalent refraction and high order aberrations ( P CONCLUSION: The success rate of SCC in T - PRK surgery is higher than that in LASIK, DCC in T - PRK surgery is higher than that in LASIK, and accurate measurement of SCC and DCC can be effective to compensate for it.

Quality standard study on Tibetan medicine Gentianae Szechenyii Flos / 中国中药杂志

In order to efficiently control the quality of the Tibetan medicine Gentianae Szechenyii Flos, the quality standard was established in this study. The tests of water content, total ash and ethanol-soluble extractives of the crude drugs were carried out based on the methods recorded in appendix of Chinese Pharmacopeia (2010 edition, volume 1). The TLC method was established by using reference drug and gentiournoside A as reference substance, and a mixture of ethyl acetate-methanol-water-formic acid (7: 1.5: 1: 0.2) as the developing solvent system on silica gel G TLC plate. The content of gentiournoside A was assayed by HPLC on a Ultimate XB-C18 (4.6 mm x 250 mm, 5 μm) column, using methanol-water (0.02% phosphoric acid) (52:48) as the mobile phase at a flow rate of 1.0 mL x min(-1). The column temperature is 25 degrees C and the detection wavelength is at 240 nm. As a result, gentiournoside A and the other constituents were separated and presented the same fluorescence light comparing with the reference substance on TLC detected under the UV light(366 nm). The methodology validation for the assay of gentiournoside A showed that it was in a good linear correlation in the range of 10.01-400.32 mg x L(-1) with the regression equation of Y = 1 539.5X - 33.339 (r = 0.999 7), and the average recovery was 99.68% (RSD 1.92%). The mass fractions of gentiournoside A, water content, ethanol-soluble extractives of 19 batches samples were varied in the ranges of 14.48-31.51 mg x g(-1), 11.25% -12.74% and 24.21% - 31.60%, respectively, and total ash was 4.64% - 6.12% detected from 10 batches samples. The recommended standards of quantitative indexes are that the mass fractions of gentiournoside A and extractives are not less than 15.0 mg x g(-1) (1.5%) and 21.0%, respectively; the water and total ash are not more than 13.0% and 6.0%, respectively.

Preparation technology of supersaturatable self-microemulsifying drug delivery system of etoposide and quality evaluation / 中草药

To prepare supersaturatable self-microemulsifying drug delivery system (S-SMEDDS) of etoposide (VP-16) for increasing the solubility of difficult soluble drug of etoposide, which will provide a scientific basis for improving its bioavailability. To study the prescription and preparation technology of S-SMEDDS of VP-16, according to different oil phases, compatibility of surfactants, and the microemulsion area size in the pseudo ternary phase diagram of different cosurfactants, to determine the basic prescription of self-microemulsifying concentrate, optimize the prescription of VP-16 based on its solubility and crystallization conditions, with filtrating appropriate precipitation inhibitor and the best prescription drug loading. The rate of self-microemulsifying was taken as index to study the preparation technology of VP-16 S-SMEDDS for investigating the influence on the ability of self-microemulsifying. The optimal prescription is: RH40-PEG 400-GTCC-PVP K30 (20∶ 20∶ 10∶ 1), 2% drug content of the mass fraction. The optimum technological conditions are 37 ℃, 20 r/min, and 20 min by magnetic stirring. The average particle size of VP-16 S-SMEDDS is (82.7 ± 3.3) nm and the size distribution of VP-16 S-SMEDDS is relatively concentrated. The average content of VP-16 in three batches of S-SMEDDS is 19.98 mg/g. Results of dissolution test showed that at 60 min the cumulative dissolution is close to 100%. Stability study results show that the high temperature and light could influence the drug stability and micro emulsification ability of VP-16 S-SMEDDS, while the psychro-thermal cycles test has no influence to it. After the preliminary stability test, the results show that the stability of VP-16 S-SMEDDS is good. The optimized prescription of VP-16 S-SMEDDS can significantly increase the solubility of VP-16, and it's quality is stable, which could improve its bioavailability further. The research method is scientific, reliable, and feasible.

Analysis of clinical characteristics and prognostic factors of primary mediastinal T-cell lymphoblastic lymphoma / 中国实验血液学杂志

Lymphoblastic lymphoma (LBL) comprises 2% to 4% of non-Hodgkin lymphomas cases in adults, of which 85% to 90% of LBL in adults is of T-cell phenotype. This study was aimed to evaluate the clinical characteristics and prognostic factors of patients with mediastinal T-LBL. Based on the retrospective analysis of the clinical data of 35 patients with mediastinal T-LBL during the period from January 1998 to January 2011, the clinical characteristics and prognostic factors of mediastinal T-LBL were summarized. The results showed that the total of 35 patients were identified (male 24 and female 11), with a median age of 19 (5 - 52) years. The majority of patients were in stage III/IV, 16 cases (45.7%) presented bulky mediastinal mass. Intrathoracic effusions (pleural, pericardial) were not uncommon (62.9%). Overall survival rate (OS) and progression-free survival rate (PFS) at 3 years for the entire cohort were 36% and 24%, respectively. OS and PFS at 5 years were 25% and 16.7%, respectively. Anemia at diagnosis were an important, independent predictor of OS (P = 0.048). Bulky mass (P = 0.048), superior vena cava syndrome (P = 0.021), and abnormal PLT count at diagnosis was the independent prognostic factors for PFS (P = 0.021). It is concluded that the patients with primary mediastinal T-LBL are characterized by a low incidence, bad prognosis, and short survival. For patients accompanying with anemia, bulky mass and superior vena cava syndrome, their prognosis is worse.

Relation of B7-H3 molecule expression in multiple myeloma with poor prognosis and bone destruction / 中国实验血液学杂志

This study was purpose to investigate the B7-H3 expression in multiple myeloma cell lines and CD138 cells of patients with multiple myeloma, and explore its clinical significance. Three myeloma cell lines (RPMI8226, U266 and H929) were used. Forty-five patients with multiple myeloma were enrolled in the study. The expression of B7-H3 was detected by flow cytometry and RT-PCR. The relationship between B7-H3 and clinical prognostic factor was analyzed. The results showed that (1)In myeloma cell lines, high expression of B7-H3 was seen in RPMI8226 (92.30 ± 1.1)% and U266 (79.03 ± 1.2)% but not in H929 cell line (4.26 ± 0.2)%. (2) Exogenous IL-6 had no effect on upregulation of B7-H3 in myeloma cell lines. (3) In multiple myeloma patients, the proportions of B7-H3 positive cells in newly diagnosed, remission and relapsed patients were (48.58 ± 33.593)%, (22.16 ± 18.853)%, and (57.65 ± 28.296)%, respectively. The difference between the newly diagnosed and remission patients, and remission and relapsed patients was significant (P = 0.023, P = 0.004). (4)High B7-H3 expression was correlated with high numbers of bone destruction and high levels of serum calcium (P = 0.027, P = 0.046, respectively). It is concluded that the relation of B7-H3 molecule expression with prognosis of multiple myeloma may be negative, but with degree of bone destruction is positive, thus the high expression of B7-H3 may correlated with disease progression and bone destruction of patients with multiple myeloma.

Rapid recognition and identification of phenanthrenes in Juncus effusus and Juncus setchuensis by LC-ESI-MS / 中草药

Objective: To rapidly recognize and identify phenanthrenes in Juncus effusus and J. setchuensis. Methods: HPLC-ESI-MS method was used to investigate the ESI-MS characteristics of those phenanthrenes, to screen, and to identify phenanthrenes in J. effusus and J. setchuensis. Results: Under negative mode, phenanthrenes were inclined to loss two hydrogens, which presumably caused by the cyclization of vinyl and aromatic ring; Furthermore, the loss of C=O could be also observed. Fragmentation pathway of phenanthrenes in ESI as a negative ion mode was summarized. Twenty-one phenanthrenes were rapidly detected from the extracts of J. effusus and J. setchuensis. Among them, six compounds were accurately identified by comparison with reference substances; The structures of five compounds were inferred according to their MS spectrum and polarity. Conclusion: HPLC-ESI-MS provides not only a new technique for the rapid identification of phenanthrenes from complex matrix, but also an effective method for the target separation of constituents in Chinese materia medica.