OBJECTIVE To analyze the types and control measures of major microorganisms in pharmaceutical water systems, so as to provide guidance for effective control of pharmaceutical water systems. METHODS The main microbial species, abundance and harmfulness of drinking water, purified water and water for injection were reviewed, and the control measures on microorganisms in pharmaceutical water were discussed. RESULTS There were differences in the main microbial types in pharmaceutical water. Burkholderia cepacia complex and Ralstonia pickettii were conditioned pathogens in pharmaceutical water, thus causing certain biological safety hazards. CONCLUSION Pharmaceutical companies can strengthen the control of microorganisms in the water system by establishing microbial databases and common microbial strain banks at all levels. Trend analysis should to be conducted based on alert limits and action limits, so as to strengthen the control of microorganisms in the water system.

ObjectiveTo investigate the effects of the volatile oil of Linderae Radix on the apoptosis and autophagy of human gastric cancer cell line AGS， and to explore the regulatory role of adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR） signaling pathway in this process. MethodThe volatile oil of Linderae Radix was extracted by steam distillation， and the effect of the volatile oil on the viability of AGS cells was detected by thiazolyl tetrazolium （MTT） colorimetry. The optimal intervention dose and time were determined according to the half maximal inhibitory concentration （IC₅₀） for subsequent research. The blank， low， medium， and high-dose volatile oil （0， 15， 30， 60 mg·L^-1） groups and the positive drug cyclophosphamide （CTX， 350 mg·L^-1） group were designed. AGS cells were treated with different doses of volatile oil for 48 h. The changes in cell proliferation， cycle， and migration were measured by colony formation assay， flow cytometry， and cell scratch test， respectively. Hematoxylin-eosin （HE） staining was employed to observe the changes of cell morphology， Annexin-V/propidium iodide （PI） double staining to measure the apoptosis， and acridine orange （AO） staining to measure the autophagy level of the cells. Western blotting was employed to determine the expression of the autophagy effectors Beclin-1， p62， microtubule-associated protein 1-light chain 3 （LC3）， B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cleaved Caspase-3， cleaved poly ADP-ribose polymerase （PARP）， adenosine monophosphate-activated protein kinase （AMPK）， phosphorylated AMPK （p-AMPK）， mTOR， and phosphorylated mTOR （p-mTOR）. ResultCompared with the blank group， 24 h and 48 h of intervention with the volatile oil of Linderae Radix inhibited the viability of AGS cells in a concentration- and time-dependent manner （P<0.05， P<0.01）. Compared with the blank group， the volatile oil decreased the cell proliferation and migration （P<0.05， P<0.01） and blocked the AGS cell cycle in G₂/M phase （P<0.05， P<0.01） in a concentration-dependent manner. The cells treated with the volatile oil became spherical and smaller， with the formation of apoptotic bodies and increased apoptosis rate （P<0.05， P<0.01）. As the dose of the volatile oil increased， the number of autophagosomes increased and the red fluorescence gradually enhanced， indicating the elevated level of autophagy. Compared with the blank group， different doses of volatile oil up-regulated the protein levels of Beclin-1， LC3 Ⅱ/LC3 Ⅰ， cleaved Caspase-3， cleaved PARP， Bax/Bcl-2， and AMPK （P<0.05， P<0.01） and down-regulated the protein levels of p62 and p-mTOR （P<0.05， P<0.01）. ConclusionThe volatile oil of Linderae Radix induces the apoptosis and exerts the autophagy-mediated growth inhibition of AGS cells by regulating the AMPK/mTOR signaling pathway.

ObjectiveTo explore the anti-inflammatory effect of Duhuo Jishengtang （DHJST） on collagen-induced arthritis （CIA） model rats and its effect on the Toll-like receptor 2 （TLR2）/p38 mitogen-activated protein kinase （MAPK）/nuclear factor-κB （NF-κB） signaling pathway. MethodForty-eight male SD rats were randomly divided into the following six groups （n=8）： normal group， model group， methotrexate （MTX） group， low-dose DHJST （DHJST-L） group， medium-dose DHJST （DHJST-M） group， and high-dose DHJST （DHJST-H） group. The CIA model was established by injecting bovine type Ⅱ collagen into the rat tail root with the collagen antibody induction method. After model induction， rats were treated with drugs by gavage. The rats in the MTX group received MTX at 2.0 mg·kg^-1， three times a week， and those in the DHJST groups received DHJST at 3.8， 7.6， 15.2 g·kg^-1·d^-1 for 28 days. The rats in the normal group and the model group were given the same dose of normal saline. The weight of the rats was recorded， and the paw swelling degree was observed. The arthritis index and immune organ index were measured， and the changes in the microcirculation indexes of the rats were detected with a microcirculation detector. Hematoxylin-eosin （HE） staining was used to detect the pathological morphologic changes in rat synovial tissues and the apoptosis rate of synovial cells was detected by flow cytometry to determine the therapeutic effect of DHJST on rheumatoid arthritis. Enzyme-linked immunosorbent assay （ELISA） was used to detect the changes in serum levels of tumor necrosis factor-α （TNF-α）， interleukin （IL）-1β， IL-17A， and interferon-γ （IFN-γ）. The protein expression of TLR2， NF-κB p65， phosphorylated NF-κB p65 （p-NF-κB p65）， p38 MAPK， and p-p38 MAPK was detected by Western blot. ResultCompared with the normal group， the model group showed reduced body weight （P<0.01）， increased paw swelling degree， arthritis index， and immune organ index （P<0.01）， increased comprehensive microvascular score and vascular resistance （P<0.01）， significant hyperplasia of synovial tissues and massive infiltration of inflammatory cells as revealed by pathological sections， and up-regulated expression levels of TNF-α， IL-1β， IL-17A， and IFN-γ in serum， and TLR2， p-NF-κB p65/NF-κB p65 and p-p38 MAPK/p38 MAPK in synovial tissues （P<0.01）. Compared with the model group， the DHJST groups showed increased body weight of rats （P<0.01）， decreased paw swelling degree， arthritis index， and immune organ index （P<0.05， P<0.01）， reduced comprehensive microvascular score and vascular resistance （P<0.05， P<0.01）， improved synovial histopathological injury， increased apoptosis rate of synovial cells （P<0.01）， and down-regulated levels of TNF-α， IL-1β， IL-17A， and IFN-γ in serum （P<0.05， P<0.01） and TLR2， p-NF-κB p65/NF-κB p65 and p-p38 MAPK/p38 MAPK in synovial tissues （P<0.05， P<0.01）. ConclusionDHJST may alleviate the inflammatory reaction in CIA rats by regulating the TLR2/p38 MAPK/NF-κB signaling pathway， thus exerting its anti-rheumatoid arthritis effect.

Exosome is a kind of vesicle secreted by a variety of cells with lipid bilayer membrane structure， which has good biocompatibility， high targeting and high stability， and is a natural nanoscale drug carrier with great development potential in drug delivery system. In this paper， exosomes and their properties， exosome drug delivery pathways and methods， the design strategy of engineered exosome drug delivery systems for targeted disease therapy， and the application of exosome drug delivery systems in the treatment of a variety of diseases were reviewed. Exosome drug delivery pathways could be divided into two categories： exogenous and endogenous. Common exosome drug delivery methods included electroporation， co-incubation， and ultrasound. Engineered exosome drug delivery system can further improve drug loading and enhance drug targeting. The main way of engineering is to modify exosome surface through genetic engineering technology， physical modification， chemical modification， etc. Exosome drug delivery system provides a new idea for targeted therapy of arthritis， tumor， brain and other diseases.

Objective:To investigate the safety of minimally invasive liver resection for resectable hepatocellular carcinoma (HCC) complicated with portal hypertension.Methods:The propensity score matching and retrospective cohort study was conducted. The clinicopathological data of 807 patients with resectable HCC who underwent minimally invasive liver resection in 8 medical centers, including Sir Run Run Shaw Hospital, Affiliated with the Zhejiang University School of Medicine et al, from June 2011 to November 2022 were collected. There were 670 males and 137 females, aged 58(50,66)years. Of the 807 patients, 173 cases with portal hypertension were divided into the portal hypertension group, and 634 cases without portal hypertension were divided into the non-portal hypertension group. Observation indicators: (1) propensity score matching and comparison of general data of patients between the two groups after matching; (2) intraoperative and post-operative situations; (3) subgroup analysis. Propensity score matching was done by the 1:1 nearest neighbor matching method, with the caliper setting as 0.001. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the rank sum test. Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Comparison of ordinal data was constructed using the non-parameter rank sun test. Results:(1) Propensity score matching and comparison of general data of patients between the two groups after matching. Of the 807 patients, 268 cases were successfully matched, including 134 cases in the portal hypertension group and 134 cases in the non-portal hypertension group. The elimination of the tumor diameter and robot-assisted surgery confounding bias ensured comparability between the two groups after propensity score matching. (2) Intraoperative and postoperative situations. The occlusion time of porta hepatis, cases with intraoperative blood transfusion, cases with postoperative complication, cases with complication >Ⅱ grade of Clavien-Dindo classification, cases of Clavien-Dindo classification as Ⅰ grade, Ⅱ grade, Ⅲ grade, Ⅳ grade, cases with liver related complication were 27.0(15.0,43.0)minutes, 33, 55, 15, 13, 29, 14, 1, 37 in the portal hypertension group, versus 35.0(22.0,60.0)minutes, 17, 25, 5, 14, 9, 4, 1, 13 in the non-portal hypertension group, showing significant differences in the above indicators between the two groups ( Z=-2.15, χ2=6.30, 16.39, 4.38, 20.72, 14.16, P<0.05). (3) Subgroup analysis. Results of subgroups analysis showed that in cases with major live resection, the operation time, volume of intraoperative blood loss, duration of postoperative hospital stay were 243.5(174.6,296.3)minutes, 200.0(150.0,600.0)mL, 7.5(6.0,13.0)days in the portal hypertension group, versus 270.0(180.0,314.5)minutes, 200.0 (75.0,450.0)mL, 7.0(5.5,10.0)days in the non-portal hypertension group, showing no significant difference in the above indicators between the two groups ( Z=-0.54, -1.73, -0.92, P>0.05). In cases with non-major live resection, the operation time, volume of intraoperative blood loss, duration of postoperative hospital stay were 170.0(120.0,227.5)minutes, 100.0(50.0,200.0)mL, 8.0(5.0,10.0)days in the portal hypertension group, versus 170.0(120.0,227.5)minutes, 100.0(50.0,200.0)mL, 7.0(5.5,9.0)days in the non-portal hypertension group, showing no significant difference in the above indicators between the two groups ( Z=-1.39, -0.10, 1.05, P>0.05). In cases with anatomical liver resection, the operation time, volume of intraoperative blood loss, duration of postoperative hospital stay were 210.0(150.0,285.0)minutes, 150.0(50.0,200.0)mL, 8.0(6.0,9.3)days in the portal hypertension group, versus 225.5(146.3,306.8)minutes, 100.0(50.0,250.0)mL, 7.0(6.0,9.0)days in the non-portal hypertension group, showing no significant difference in the above indica-tors between the two groups ( Z=-0.75, -0.26, -0.91, P>0.05). In cases with non-anatomical liver resection, the operation time, volume of intraoperative blood loss, duration of postoperative hospital stay were 173.5(120.0,231.5)minutes, 175.0(50.0,300.0)mL, 7.0(5.0,11.0)days in the portal hyper-tension group, versus 186.0(123.0,262.5)minutes, 100.0(50.0,200.0)mL, 7.0(5.0,9.5)days in the non-portal hypertension group, showing no significant difference in the above indicators between the two groups ( Z=-0.97, -1.12, -0.98, P>0.05). Conclusion:Minimally invasive liver resection or even major liver resection is safe and feasible for screened HCC patients complicated with portal hyper-tension, but attention should be paid to the prevention and treatment of postoperative complications.

Objective:To investigate the perioperative efficacy of robot surgical system assisted anatomic and non-anatomic hepatectomy.Methods:The propensity score matching and retrospective cohort study was conducted. The clinical data of 103 patients who underwent robot surgical system assisted hepatectomy in Sir Run Run Shaw Hospital, Affiliated with the Zhejiang University School of Medicine from March 2016 to December 2021 were collected. There were 54 males and 49 females, aged 56(range, 44?64)years. Of the 103 patients, 55 cases undergoing robot surgical system assisted anatomic hepatectomy were divided into the anatomic group, and 48 cases undergoing robot surgical system assisted non-anatomic hepatectomy were divided into the non-anatomic group. Observation indicators: (1) propensity score matching and comparison of general data of patients between the two groups after matching; (2) intraoperative conditions; (3) perioperative complications. Propensity score matching was done by the 1:1 nearest neighbor matching method. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the t test. Measurement data with skewed distribution were expressed as M(range), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Comparison of ordinal data was conducted using the rank sum test. Results:(1) Propensity score matching and compari-son of general data of patients between the two groups after matching. Of the 103 patients, 94 cases were successfully matched, including 47 cases in the anatomic group and 47 cases in the non-anatomic group. The elimination of preoperative body mass index, preoperative platelet and preoperative albumin confounding bias ensured comparability between the two groups after propensity score matching. (2) Intraoperative conditions. After propensity score matching, the operation time and volume of intraoperative blood loss were 175(range, 120?240)minutes and 50(range, 50?100)mL in patients of the anatomic group, versus 155(range, 105?190)minutes and 100(range, 50?200)mL in patients of the non-anatomic group, showing significant differences in the above indicators between the two groups ( Z=1.97, 2.49, P<0.05). (3) Perioperative complications. After propensity score matching, cases with pleural fluid and/or ascites, case with biliary fistula, case with thrombosis, case with peritoneal infection, case with incision infection were 11, 1, 2, 4, 1 in patients of the anatomic group, versus 12, 0, 4, 1, 0 in patients of the non-anatomic group, showing no significant difference in the above indicators between the two groups ( P>0.05). Cases with complications classified as grade Ⅰ, grade Ⅱ, grade Ⅲ, grade Ⅳ of the Clavien-Dindo classification were 33, 14, 0, 0 in patients of the anatomic group, versus 28, 14, 3, 2 in patients of the non-anatomic group, showing no significant difference in the above indicators between the two groups ( Z=?1.38, P>0.05). Conclusions:Robotic surgical system assisted anatomic and non-anatomic hepatectomy are safe and feasible for clinical application. Compared with robot surgical system assisted non-anatomic hepatectomy, patients under-going robot surgical system assisted anatomic hepatectomy have long operation time and less volume of intraoperative blood loss.

OBJECTIVE To investigate the effect and potential mechanism of safflower polysaccharide on apoptosis and autophagy of human hepatocellular carcinoma cells. METHODS Human hepatocellular carcinoma HepG2， SMMC-7721 and Huh-7 cells were selected as subjects， and safflower polysaccharide was used as intervention drug to screen sensitive cells， intervention concentration and intervention time. The sensitive cells were selected as the object and intervened with different concentrations of safflower polysaccharide； the apoptosis， migration， clone formation， morphology and autophagy of human hepatocellular carcinoma cells were observed； the expressions of apoptosis， autophagy and phosphatidyl inositol-3-kinase （PI3K）/protein kinase B （Akt）/mammals rapamycin target protein （mTOR） signaling pathway related protein were detected. RESULTS safflower polysaccharide could inhibit the proliferation of 3 kinds of human hepatocellular carcinoma cells， and the half inhibition concentration of it to SMMC-7721 cells was significantly lower than to other two kinds of cells （P＜0.05）. After 48 h intervened with low， medium and high concentrations of safflower polysaccharide （20， 40， 80 μmol/L）， the apoptosis of SMMC-7721 cells was increased compared with the control group， and cell migration rates at 24 and 48 h （except for safflower polysaccharide low- dose group at 24 h） and clone formation rate at 24 h were significantly decreased compared with the control group （P＜0.05 or P＜ 0.01）. Compared with the control group， cell number in safflower polysaccharide groups was significantly decreased， and autophagy levels were improved to some extent； the relative expressions of cleaved caspase-3， caspase-9， Bax and beclin-1 protein and ratio of LC3Ⅱ/LC3Ⅰ were significantly increased， and the relative protein expressions of Bcl-2， p62， PI3K， mTOR and Akt were significantly decreased （P＜0.05 or P＜0.01）. CONCLUSIONS Safflower polysaccharide could effectively inhibit the proliferation and induce apoptosis and autophagy of human hepatocellular carcinoma SMMC-7721 cells， the mechanism of which may be related to the inhibition of PI3K/Akt/mTOR signaling pathway.

In this study, the necessity of professional master's degree training in clinical pharmacy was analyzed by means of literature research and practice summary, and the existing problems in current professional curriculum, tutor team and practice teaching were discussed. In view of the existing problems, this paper puts forward that medical colleges and universities should give full play to their medical resources, draw lessons from the talents training experience of clinical pharmacists training base, adopt measures such as optimizing curriculum system, selecting tutors, attaching importance to practical teaching, so as to improve the quality of postgraduate training in clinical pharmacy, train high-level clinical pharmacists, and promote the development of clinical pharmacy in China.

Intrahepatic biliary cystadenoma (IBC) is a rare benign cystic neoplasm of liver,with malignant potential to transform into intrahepatic biliary cystadenocarcinoma.IBC predominantly occurs in women up to 85％,showing no special clinical symptoms and a polycystic lesion inside the liver on image examination.It is difficult to differentially diagnose from other cystic lesions of the liver such as simple liver cyst,intraductal papillary neoplasm of the bile duct,etc.The missed diagnosis and misdiagnosis rate were rather high in IBC,which is accurately diagnosed by pathological examination.Complete surgical removal of the tumor is the best way to cure it,and also can bring a satisfactory outcome to patients.

OBJECTIVETo observe effects of on hepatocarcinoma (HCC) cell vasculogenic mimicry (VM) and explore the molecular mechanism by which inhibits HCC metastasis and invasion.

METHODSForty male SD rats were randomly divided into 4 groups for gastric lavage of normal saline or high, moderate or low doses of (twice daily) for 4 consecutive days. The sera were collected from the rats for treatment of cultured human HCC HepG2 cells. VM formation in the cells was detected using an image acquisition and analysis system 24 h after incubation of the cells with the sera and with the RhoA/ROCK inhibitor Y-27632(P). The expression levels of RhoA and ROCK1 in the cells were detected using Western blotting, and the contents of VE-cadherin and PI3K in the culture supernatant were determined using ELISA.

RESULTSTreatment with the sera from -treated rats significantly inhibited formation of VM in HepG2 cells, and the diameters of VM formed were significantly greater than those in the positive control group ( < 0.01). Y-27632 completely inhibited the formation of VM in HepG2 cells ( < 0.01). Treatments with and Y-27632 both inhibited the expression of RhoA and ROCK1 ( < 0.05) and significantly lowered the contents of VE-cadherin and PI3K in the culture supernatant ( < 0.05).

CONCLUSIONS can inhibit the formation of VM in HCC cells possibly by inhibiting the RhoA/ROCK pathways and the expressions of VE-cadherin and PI3K.