Objective To investigate the silencing effect of RNA interference on MSTN gene ( myostatin, MSTN) expression, and detect the effects on the downstream genes in Schizopygopsis pylzovi. Methods To construct the recombi?nant adenovirus vector 1P3 (DSP MSTN 273+250+1737) and 1P2 (DSP MSTN 195+ 1670) for RNA interference of the MSTN gene in Schizopygopsis pylzovi, and to conduct the RNA interference in vivo experiment by injecting the vector in?to the muscle tissue of Schizopygopsis pylzovi. Real?time PCR and Western blotting were used to evaluate the silencing effects on MSTN gene expression, and to detect the regulatory function of M?CK at gene transcription level after RNA inter?ference of the MSTN gene. Results The result of real?time PCR showed that compared with the HK team ( Virus general negative control group) and N team (blank control group), the 1P3 had significant interference effect on the MSTN gene transcription in Schizopygopsis pylzovi (P<0?05), with an inhibition rate of 53?5%, but the 1P2 had no significant inter?ference effect on the MSTN gene transcription. The result of Western blotting was consistent with the results of real?time PCR. At the same time, after the 1P3 interference, the level of MSTN gene transcription was declined, and the level of M? CK gene expression was significantly increased. Conclusions Our results demonstrate that the expression of MSTN gene can be effectively suppressed, and the expression of M?CK gene can be up?regulated through the RNA interference. There?fore, it proves that MSTN gene can inhibit the transcription of M?CK gene in Schizopygopsis pylzovi, and reveals the regula?tory role of MSTN gene in the muscle growth and development in the plateau fish Schizopygopsis pylzovi.

OBJECTIVE:To explore the effects and mechanism of extracts,active constituents and constituent combination of Sinopodophylli Fructus on cell proliferation of human breast cancer. METHODS:Acid phosphatase method was conducted to deter-mine the effects of 4 extracts [ethanol extract (Xc),petroleum ether extract from ethanol extract (Xp),ethyl acetate extract from ethanol extract (Xe),n-butanol extract from ethanol extract (Xz)],5 active constituents [podophyllotoxin (S1),deoxypodophyllo-toxin (S2),4-desmethyl deoxypodophyllotoxin (S3),8-isopentenyl kaempferol (S4),8,2′-diisoprenyl quercetin-3-methyl ether (S5)] and 3 active constituent combination [combination 1,S1-S2-S3-S4-S5 (2:4:1:4:32),Z1;combination 2,S2-S4 (1:1),Z2;combination 3,S3-S4(1:4),Z3] on the MDA-MB-231,MCF-7 cell proliferation;flow cytometry was adopted to detect the effects of above-mentioned samples on MDA-MB-231,MCF-7(T47D)cell cycle and mitochondrial membrane potential. RESULTS:The active constituent combination Z1 showed significant inhibitory effects on MDA-MB-231,MCF-7 cells,the half inhibitory concen-trations(IC50)were(0.27±0.2),(0.11±0.1)μg/mL;extracts Xc,Xp,Xe,active constituents S2,S4 and active constituent combi-nation Z2,Z3 showed relatively strong inhibitory effects on MDA-MB-231,MCF-7 (T47D) cell proliferation (IC50<15 μg/mL). Both extracts and active constituents can block MDA-MB-231,MCF-7 cell cycle in G2/M phase;all active constituents can block MDA-MB-231,T47D cell cycle in G0/G1 phase,and can reduce MDA-MB-231,T47D cell mitochondrial membrane potential. CONCLUSIONS:The active constituents and constituent combination of Sinopodophylli Fructus can inhibit cell proliferation of breast cancer by affecting cell cycle and mitochondrial mem-brane potential.

Objective To compare and analyze the formula composition based on their herbal nature for wind, cold, and dampness arthralgia. Methods The ancient formulas for wind, cold, and dampness arthralgia were searched and the database was established. The top 30 high-frequency herbs were analyzed with frequency, hypothesis testing and association rules. The nature, taste, and meridian distribution were used as the variable quantities for clustering analysis. Results Totally 338 formulas were collected, including 122 formulas for wind arthralgia, 110 formulas for cold arthralgia, and 106 formulas for dampness arthralgia. There are 21 same herbs among the top 30 high-frequency herbs;Tonic herbs were the highest frequently used, followed by the divergence of cold herbs in wind arthralgia, interior-warming herbs in cold arthralgia, and damp-resolving herbs in dampness arthralgia. The frequently used herbs in each kind of formula compared with other two kinds were:Cinnamomi Cortex, Aconiti Lateralis Radix Praeparata, Angelicae Sinensis Radix, Achyranthis Bidentatae Radix, Notopterygii Rhizome et Radix, Paeoniae Radix and Astrragali Radix in formulas for cold arthralgia, Poria and Atractylodis Macrocephalae Rhizoma in formulas for dampness arthralgia. Three kinds of formulas are given priority to slight warm, followed by warm and neutral. Classification is clear when most of the formulas were clustered into five classes according to their herbal nature in each kind of formula. Conclusion The three kinds of formulas cross each other but with own characteristics. A variety of data mining methods can be used to analyze scientific connotation of therapeutic principle for arthralgia.

Objective To investigate the value of serum hepatitis B virus(HBV)RNA for predicting hepatitis B virus e antigen(HBeAg)clearance and seroconversion after nucleoside(acid)analogs(NAs)treatment in patients with chronic hepatitis B(CHB).Methods Serum samples were collected from 178 CHB patients who received NAs monotherapy in Weihai Hospital affiliated to Shandong University of Traditional Chinese Medicine from February 12,2017 to February 21,2019.Serum HBV RNA levels were analyzed by real-time fluorescence-based quantitative PCR at baseline and after NAs treatment.Results The patients with HBeAg clearance and seroconversion showed significantly lower serum HBV RNA levels at baseline,6 months and 12 months of treatment compared with those without HBeAg clearance and seroconversion(P＜0.001).During follow-up,the patients with lower baseline HBV RNA levels had higher rate of cumulative HBeAg clearance(Log Rank x2=11.282,P=0.001)or seroconversion(Log Rank x2=10.739,P=0.001)than the patients with higher baseline HBV RNA levels.Cox regression model analysis indicated that serum HBV RNA levels at baseline,6 months and 12 months of treatment were an independent predictor for HBeAg clearance and seroconversion in CHB patients(P＜0.05).The area under the ROC curve(AUC)of baseline serum HBV RNA level was 0.808(95％CI:0.743-0.872)for predicting HBeAg clearance and 0.824(95％CI:0.763-0.885)for predicting seroconversion.The AUC of HBV RNA level at 6 months of treatment was 0.830(95％CI:0.765-0.894)for predicting HBeAg clearance and 0.732(95％CI:0.657-0.808)for predicting seroconversion.The AUC of HBV RNA level at 12 months of treatment was 0.737(95％CI:0.641-0.833)for predicting HBeAg clearance and 0.757(95％CI:0.671-0.842)for predicting seroconversion.The AUC of baseline HBV RNA level combined with HBV RNA decline at 6 months of treatment was 0.856(95％CI:0.795-0.917)for predicting HBeAg clearance and 0.864(95％CI:0.802-0.926)for predicting seroconversion.The AUC of baseline HBV RNA level combined with HBV RNA decline at 12 months of treatment was 0.881(95％CI:0.826-0.936)for predicting HBeAg clearance and 0.848(95％CI:0.784-0.911)for predicting seroconversion.Conclusions Serum HBV RNA levels have high predictive value for HBeAg clearance and seroconversion after NAs therapy in CHB patients,suggesting that HBV RNA levels are helpful for identifying non-responders and informing combination therapy.

ObjectiveTo analyze the epidemiological characteristics of scarlet fever in Putuo District， Shanghai from 2015 to 2020， and provide a basis for formulating effective prevention and control measures. MethodsThe data of scarlet fever cases from January 1， 2015 to December 31， 2020 in Putuo District， Shanghai were obtained from the Infectious Disease Reporting Information Management System of the Chinese center for disease control and prevention （CDC）. Descriptive epidemiological method was used to analyze the epidemiological characteristics of scarlet fever cases in Putuo District， Shanghai by time， population， and region. ResultsA total of 586 scarlet fever cases were reported in Putuo District from 2015 to 2020， with an average annual reported incidence of 7.39 per 10⁵， an average age of onset of 6.2±1.8 years， and a male/female ratio of 1.6∶1. The incidence of scarlet fever has declined significantly in 2020， which was 5.44 per 10⁵ lower than the average reported incidence in 2015‒2020. The incidence peak was from April to June and November to January of the following year， which was characterized by high incidence in spring and winter. The age distribution of cases was concentrated at the age of 4‒9 years， mainly children in kindergarten and primary school students. Areas with high reported incidence rates were Wanli Street， Changzheng Town， Taopu Town and Changfeng New Village Street. ConclusionThe peak incidence of scarlet fever in Putuo District is from April to June and November to January of the following year， with children in kindergartens and students in primary school as the key populations. We should strengthen epidemic monitoring and guidance of prevention and control for in kindergartens and schools before the high incidence season of scarlet fever， so as to effectively prevent the epidemic of the disease.

The mechanisms underlying autophagic defects in nonalcoholic steatohepatitis (NASH) remain largely unknown. We aimed to elucidate the roles of hepatic cyclooxygenase 1 (COX1) in autophagy and the pathogenesis of diet-induced steatohepatitis in mice. Human nonalcoholic fatty liver disease (NAFLD) liver samples were used to examine the protein expression of COX1 and the level of autophagy. Cox1^Δhepa mice and their wildtype littermates were generated and fed with 3 different NASH models. We found that hepatic COX1 expression was increased in patients with NASH and diet-induced NASH mice models accompanied by impaired autophagy. COX1 was required for basal autophagy in hepatocytes and liver specific COX1 deletion exacerbated steatohepatitis by inhibiting autophagy. Mechanistically, COX1 directly interacted with WD repeat domain, phosphoinositide interacting 2 (WIPI2), which was crucial for autophagosome maturation. Adeno-associated virus (AAV)-mediated rescue of WIPI2 reversed the impaired autophagic flux and improved NASH phenotypes in Cox1^Δhepa mice, indicating that COX1 deletion-mediated steatohepatitis was partially dependent on WIPI2-mediated autophagy. In conclusion, we demonstrated a novel role of COX1 in hepatic autophagy that protected against NASH by interacting with WIPI2. Targeting the COX1-WIPI2 axis may be a novel therapeutic strategy for NASH.