Objective We investigated if the proliferative capacity of endothelial progenitor cells was affected by hypoxia and exercise. Methods Twenty four male Sprague-Dawley rats were randomly and averagely divided into 4 groups: (1) living at low altitude (LL), (2) living and training at low altitude (LLTL), (3) living at high altitude (LH), and (4) living at high altitude and training at low altitude (LHTL). Eight-week incremental treadmill exercise and hypoxic simulation were used to establish LHTL animal model. Mononuclear cells from peripheral blood were obtained by density gradient centrifugation 12 hours by the end of 8-week experiment. The cells were suspended in conditioned medium 199 for culturing in vitro. Their phenotypes were confirmed by uptake of acetylated LDL and binding of fluoresce in isothiocyanate (FITC)-labeled Ulex europaeus agglutinin 1 (UEA-1) lectin. Inverted microscopic observation was used to identify the morphological changes in endothelial progenitor cells and measure the cell count. Results Adherent cells and early CFUs in groups LLTL, LH and LHTL increased more obviously than in group LL(P<0.01), whereas the number of attached cells between group LH and LHTL was not different (P>0.05). Conclusion Proliferative capacity of endothelial progenitor cells can be promoted by both hypoxic stimulation and exercise, and the promotion is more significant if combination of hypoxia and exercise was employed simultaneously.

Objective To analyze whether studying abroad has any influence on clinicians′scientific research ability, and study the rationality of the indicators reflecting the scientific research ability. Methods A total of 36 clinicians who studied abroad more than 3 months between 2008 and 2010 were selected as the abroad group, and the control group of 36 clinicians was selected from the clinician information database of the hospital by the method of Propensity Score Match.A comparison and analysis were made regarding the number of papers(SCI), grants and scientific and technological achievements between the two groups. Meanwhile, covariance analysis was used to compare the difference in the number of scientific research achievements between the two groups.Logistic regression analysis was used to compare the changes in scientific research achievements of clinicians before and after their abroad study. Results The number of articles( SCI) published and grants obtained in the 3 years after going abroad was significantly higher than that of the control group, and the number of grants obtained in the 4-6 years after going abroad was also significantly higher than that in the control group.The number of papers(SCI)published in the three years after going abroad was more than that in the 3 years before going abroad.The number of grants obtained in the 4-6 years after going abroad was more than that in the 3 years before going abroad.These differences were statistically significant. Conclusions Studying abroad has a beneficial effect on the improvement of clinicians′ scientific research ability. It is reasonable and practical to take the number of SCI articles published as the indicator of short-term changes and the number of funds obtained as the indicator of long-term changes in scientific research ability.

OBJECTIVETo examine the correlation between serum human epithelial growth factor receptor 2 extracellular domain (HER2 ECD) and circulating tumor cells (CTC), as well as the dynamic variation of HER2 ECD and its correlation to the therapeutic efficacy.

METHODSFifty-three advanced gastric cancer (AGC) patients who treated in Peking University Cancer Hospital and ever enrolled into CTC study (ClinicalTrial gov. ID: NCT01625702) were retrospectively included in this study.

INCLUSION CRITERIAthe patients were histologically confirmed as locally advanced or recurrent and/or metastatic adencarcinoma; they received two or more cycles of fluorouracil-based chemotherapy or combination targeted therapy; serum CTC was counted before and after therapy; the clinical response was evaluated every 2 cycles of treatment by the presence of at least one measurable lesion according to RECIST version 1.1 criteria. This study was approved by Ethics Committee of Peking University Cancer Hospital, and informed consents were signed by patients. The sera before and after two cycles of treatment were collected for CTC enumeration and HER2 ECD detection, in which the levels of HER2 ECD were measured by chemiluminescence immunoassays method. The positive threshold value of HER2 ECD and CTC number were ≥15 μg/L and ≥3 CTCs/7.5 ml respectively. The progression-free survival (PFS) and overall survival (OS) were compared among different groups using Log-rank tests.

RESULTSIn 53 enrolled patients, 39 were histologically identified as negative HER2, 9 as positive HER2 and another 5 cases were unknown. All the patients received fluorouracil-based chemotherapy, and 9 positive HER2 patients received combined anti-HER2 targeted therapy. Before therapy, the median HER2 ECD concentration of 53 cases was 10.45 (8.0 to 83.2) μg/L. Seven patients exhibited positive HER2 ECD levels, in whom 4 were histologically HER2 positive, but 3 were histologically HER2 negative. The median CTC number of 53 cases was 2 (0 to 668) CTCs/7.5 ml, and the positive rate of CTC was 47.2%(25/53). Following 2 cycles of therapy, a total of 10 histologically HER2 negative patients exhibited positive HER2 ECD levels, in whom 2 also possessed positive HER2 ECD levels, 83.3 μg/L and 46.9 μg/L before therapy, and 22.4 μg/L and 20.4 μg/L after therapy respectively, whereas another 8 patients (10.3 to 14.5 μg/L before therapy) acquired the elevated expression of HER2 ECD following therapy (15.1 to 19.5 μg/L). It seems that the increased level of HER2 ECD after therapy was, though not statistically significant, correlated to low number of CTCs. In histologically HER2 negative patients, pretherapeutic HER2 ECD level (positive vs. negative) was not significantly correlated to PFS (7.6 months vs. 4.4 months, P=0.328) and OS (13.6 months vs. 10.9 months, P=0.679). However, in histologically HER2 positive patients, patients with positive HER2 ECD level before therapy exhibited longer PFS (10.7 months vs. 4.2 months, P=0.025) and OS (16.5 months vs. 8.9 months, P=0.015) compared to those with negative HER2 ECD level. Additionally, CTC number was significantly correlated to prognosis in histologically HER2 negative patients. Patients with positive pretherapeutic CTC number showed longer PFS (5.3 months vs. 3.3 months, P=0.049) and OS (14.3 months vs. 7.6 months, P=0.001) as well. While in histologically HER2 positive patients, CTC number was not obviously correlated to the PFS and OS. In above 8 negative HER2 patients acquiring elevated expression of HER2 ECD following therapy, the increased HER2 ECD level was not correlated to PFS and OS (all P>0.05). In 9 histologically HER2 positive patients, 4 patients who exhibited decreased HER2 ECD level and reduced or constant CTC number had longer PFS (7.5 to 15.3 months) and OS (11.0 to 26.3 months) compared with those 2 patients who suffered from acquired HER2 ECD level following therapy (PFS 3.0 to 4.8 months and OS 7.3 to 8.6 months).

CONCLUSIONSIn histologically HER2 positive patients, increased pretherapeutic HER2 ECD level predicts better prognosis. The acquired elevated HER2 ECD level following therapy is correlated to inefficient therapeutic response. The acquirement of elevated HER2 ECD level can also be found in histologically HER2 negative patients, which may be correlated to the corresponding variation of CTC number.

Objective:To investigate the diagnosis and management of partial or complete hydatidiform mole with coexistent intrauterine pregnancy.Methods:Clinical data of 10 cases of hydatidiform mole with coexistent intrauterine pregnancy admitted to the Third Affiliated Hospital of Guangzhou Medical University, from September 2009 to May 2019 were retrospectively described.Results:(1) During the same period, 65 960 women were delivered at our hospital, and hydatidiform mole with coexistent intrauterine pregnancy was accounted for 1/6 596, among which complete hydatidiform mole and coexisting fetus (CHMCF) and partial hydatidiform mole and coexistent fetus (PHMCF) were found in four and six cases, respectively. The mean age of the ten patients were (30.9±4.1) years old, ranging from 26 to 35 years old, with 2.5 (1-4) times of pregnancies. Nine cases were identified at 22 +3 (12 +3-32 +3) gestational weeks and one at 9 + weeks. (2) Recurrent vaginal bleeding during pregnancy occurred in six cases, nausea and vomiting in three cases, and hyperthyroidism in mid- and late pregnancy in two cases. One patient developed preeclampsia and one case of severe mitral regurgitation with mild pulmonary hypertension. (3) In the 10 patients, the summit serum β -hCG level was 139 935 (16 990-546 033) U/L, and CHMCF and PHMCF patients were 212 500 (200 000-546 033) U/L and 60 768 (16 990-225 000) U/L, respectively. (4) The ultrasound results revealed a dark honeycomb area of the placenta in five cases, placental thickening in two cases, and vesicular placenta in one case. One case was found with bilateral giant luteinized ovarian cyst by ultrasound, multiple metastases in the left lower lobe of the lung by chest CT, multiple nodules in the pleural wall of the left lung by lung MRI, and CHMCF by pelvic MRI. In one case, ultrasound at 14 weeks of gestation showed interrupted fetal abdominal wall, visible mass, gastric bubble, liver, part of the intestinal echoes, and omphalocele. One case was found with embryo arrest. (5) The karyotype analysis of one case through amniocentesis was 46,XX with no anomalies, and chromosome microarray analysis was arr[hg19](1-22)×2. Prenatal diagnosis was refused in the remaining cases. (6) Among the ten patients, three were terminated by rivanol intra-amniotic injection, two received drug abortion, and uterine evacuation, and two with spontaneous abortion followed by curettage with a visible fetus and hydatidiform tissue. Total hysterectomy was performed in one patient due to partial invasion of the uterus by hydatidiform mole. One patient underwent a cesarean section on account of the left lower lung metastasis. One case developed preeclampsia at 33 +4 weeks of gestation and delivered two premature infants by cesarean section. Pathology examination found a complete and partial vesicular fetal mass in four and six cases, with P57 (－) and P57 (+), respectively. (7) During the follow-up, two women developed the persistent trophoblastic disease and received chemotherapy, while the remaining eight cases did not. Conclusions:When hydatidiform mole with coexistent intrauterine pregnancy is found, a timely differential diagnosis between CHMCF and PHMCF is needed. CHMCF is at a higher risk of abortion, intrauterine death, premature delivery, preeclampsia, and other maternal complications. Therefore, termination of CHMCF should be individualized. Most PHMCF patients have fetal malformation or fetal loss; thereby, timely termination is recommended.

Ischemic stroke is one of the leading causes of death and disability worldwide. In Han dynasty， HUA Tuo proposed the original preventive medicine idea that "with good blood circulation， the disease cannot be born"， which opened a broad space for the cross-research of blood-related mechanical factors and pharmacology. In the pathogenesis of ischemic stroke， mechanical factors comprehensively affect the function and crosstalk of platelets and endothelial cells. In recent years， as the well-known effects on thrombosis and stroke， more attention has been paid to hemodynamic factors as the participants involved in pathological mechanisms and potential therapeutic targets of ischemic stroke. The mechanical force ion channel Piezo1 widely exists on the surface of many types of cells. Besides being regulated by chemical and endogenous substances， Piezo1 responds to different mechanical conditions， regulates the opening and closing of channels， and activates different downstream signaling pathways. Piezo1 is now regarded as an important connection between mechanical and biochemical signals. A variety of Chinese medicine can affect the activity of Piezo1 protein， which may prevent and treat thrombotic diseases such as ischemic stroke through Piezo1 protein. In this paper， the effects of Piezo1 protein on the physiological and pathological functions of endothelial cells and platelet under different mechanical conditions and the role of Piezo1 in the process of thrombosis were reviewed， as well as the effects of Chinese medicine， chemical medicine， and endogenous substances targeting Piezo1 channel. These could provide new ideas for further exploring the mechanisms of Chinese medicines in activating blood circulation， developing new drugs， and deepening biomechanical-pharmacology research.

‍Traditionally， the progression from compensated liver cirrhosis to decompensated liver cirrhosis has been considered an irreversible point in the natural history of the disease； however， with the suppression of underlying etiology， cure， and disease regression， this view is challenged by an increasing number of new evidence， and the idea of “recompensation of liver cirrhosis” is gradually being accepted. In recent years， scholars in China and globally have been exploring the specific definition of recompensation of liver cirrhosis and the clinical features of patients. By summarizing the recent studies on recompensation of liver cirrhosis in China and globally， integrating existing views， and analyzing related research evidence， this article points out the main challenges in the field of recompensation at this stage， including the lack of in－depth clinical and basic research， the need to define recompensation in the context of NAFLD， and related ethical issues， in order to provide new directions for future research in this field.

‍Nonalcoholic fatty liver disease （NAFLD） is a group of highly heterogeneous diseases closely associated with metabolic dysfunction. Sarcopenia is a syndrome caused by a continuous decline in muscle mass， strength， and function， and it is often accompanied by NAFLD. Insulin resistance is the main pathological mechanism for sarcopenia and NAFLD， and in addition， factors such as changes in proteins and branched‍-‍chain amino acid， hyperammonemia， intestinal flora， and endocrine dysfunction can also lead to sarcopenia and NAFLD. With the deepening of clinical research， many published prospective studies have confirmed the existence of a bidirectional and complex pathophysiological relationship between sarcopenia and NAFLD. This article reviews the bidirectional relationship between sarcopenia and NAFLD， discusses the common pathogenesis of sarcopenia and NAFLD， summarizes the challenges faced in this field， and proposes new directions for the research on the bidirectional relationship between NAFLD and sarcopenia.

ObjectiveTo investigate the effect of Shugan Quyu Jiedu prescription （SGQYJDF） on inducing ferroptosis in hepatocellular carcinoma cells based on the tumor protein 53 （p53）/solute carrier family 7 member 11 （SLC7A11）/glutathione peroxidase 4 （GPX4） pathway. MethodMHCC97H cells were divided into the blank serum group （10% blank serum medium）， SGQYJDF-containing serum low concentration group （5% SGQYJDF-containing serum and 5% blank serum medium）， SGQYJDF-containing serum medium concentration group （7.5% SGQYJDF-containing serum and 2.5% blank serum medium）， SGQYJDF-containing serum high concentration group （10% SGQYJDF-containing serum medium） and sorafenib group （sorafenib concentration of 10 μmol·L^-1 in 10% blank serum medium）. After 24 hours of intervention， the cell survival rate was detected by cell counting kit-8 （CCK-8） assay. The cell proliferation ability was detected by 5-ethynyl-2′-deoxyuridine （EdU） staining. The intracellular ferrous ion （Fe²⁺） level was detected by ferrous ion fluorescent probe （FerroOrange） staining. The intracellular malondialdehyde （MDA） and glutathione （GSH） levels were detected by colorimetric assays. The ultrastructure of mitochondria was observed by transmission electron microscopy. The expression levels of ferroptosis-related proteins p53， SLC7A11 and GPX4 were detected by Western blot. ResultIn terms of cell viability， compared with the blank serum group， the SGQYJDF group showed a dose-dependent decrease in the survival rate of MHCC97H cells. Effect of the medium and high concentrations of SGQYJDF on the survival rate of MHCC97H cells were significantly decreased （P<0.01）. Additionally， the results of the EdU assay showed that both the medium and high concentrations of SGQYJDF were able to inhibit the proliferation ability of MHCC97H cells （P<0.05， P<0.01）. Regarding the biochemical indicators of ferroptosis， compared to the blank serum group， the medium and high concentrations of SGQYJDF were able to dose-dependently increase the intracellular Fe²⁺ level （P<0.01）. The low， medium， and high concentrations of SGQYJDF were able to dose-dependently decrease the level of GSH in MHCC97H cells （P<0.01） and increase the level of MDA in the cells （P<0.05， P<0.01）. In terms of pathway-related protein expression， compared to the blank serum group， the medium and high concentrations of SGQYJDF could significantly increase the expression of p53 （P<0.01）. The low， medium， and high concentrations of SGQYJDF could significantly decrease the expression of GPX4 （P<0.01）. The high concentration of SGQYJDF could decrease the expression of SLC7A11 （P<0.01）. In terms of the cell morphology of ferroptosis， compared with the blank serum group， transmission electron microscopy revealed that the low concentration of SGQYJDF caused mitochondrial deformation， while the medium and high concentrations of SGQYJDF resulted in reduced mitochondrial volume， increased double-layer membrane density， and decreased mitochondrial cristae. These features were similar to those of sorafenib-induced ferroptosis. Furthermore， compared with the sorafenib group， the high concentration of SGQYJDF showed no statistically significant differences in cell survival rate， proliferation ability， Fe²⁺ level， MDA level， and GSH level. ConclusionThe results suggest that SGQYJDF may induce ferroptosis and inhibit proliferation in hepatocellular carcinoma MHCC97H cells by upregulating the expression of p53， suppressing the expressions of GPX4 and SLC7A11， downregulating the level of GSH， and leading to the accumulation of intracellular Fe²⁺ and MDA.

ObjectiveTo investigate the mechanisms of Panax notoginseng saponins （PNS） in inhibiting high shear-induced platelet aggregation and thrombosis via the Piezo1-mediated calcium signaling pathway. MethodBioflux1000z was used for the microfluidic assay， where platelets were stimulated with physiological shear rate （500 s^-1）， pathological shear rate （12 000 s^-1）， or Piezo1 agonist Yoda1 under the physiological shear rate （500 s^-1）. The shear-induced platelet calcium influx and the binding of platelet with von Willebrand factor （vWF） were measured by flow cytometry. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the vWF release from platelets. The microfluidic channels were used to determine the vWF-mediated platelet aggregation and integrin αⅡbβ3 activation. A mouse model of arterial thrombosis induced by high shear stress combined with endothelial injury was established. The ultrasonic Doppler flow meter was used to monitor the cyclic flow reduction （CFR） caused by the repeated formation and shedding of thrombi， and flow cytometry was employed to examine platelet-vWF binding， on the basis of which the effect of PNS on high shear-induced arterial thrombosis was evaluated. ResultThe microfluidic assay showed that PNS decreased the high shear rate （12 000 s^-1） or Yoda1-induced calcium influx， platelet-vWF binding， vWF-mediated platelet-fibrinogen binding， and vWF release from platelet alpha-granules in a dose-dependent manner. In the mouse model of high shear-induced thrombosis， PNS markedly reduced the CFR and occlusion time of the common carotid artery and inhibited platelet-vWF binding. ConclusionPNS can mitigate pathological shear-induced platelet aggregation and arterial thrombosis via influencing Piezo1/GPIbα-vWF signaling.

OBJECTIVE: To prepare customized porous silicone orbital implants using embedded 3D printing and assess the effect of surface modification on the properties of the implants. METHODS: The transparency, fluidity and rheological properties of the supporting media were tested to determine the optimal printing parameters of silicone. The morphological changes of silicone after modification were analyzed by scanning electron microscopy, and the hydrophilicity and hydrophobicity of silicone surface were evaluated by measuring the water contact angle. The compression modulus of porous silicone was measured using compression test. Porcine aortic endothelial cells (PAOECs) were co-cultured with porous silicone scaffolds for 1, 3 and 5 days to test the biocompatibility of silicone. The local inflammatory response to subcutaneous porous silicone implants was evaluated in rats. RESULTS: The optimal printing parameters of silicone orbital implants were determined as the following: supporting medium 4% (mass ratio), printing pressure 1.0 bar and printing speed 6 mm/s. Scanning electron microscopy showed that the silicone surface was successfully modified with polydopamine and collagen, which significantly improved hydrophilicity of the silicone surface (P < 0.05) without causing significant changes in the compression modulus (P > 0.05). The modified porous silicone scaffold had no obvious cytotoxicity and obviously promoted adhesion and proliferation of PAOECs (P < 0.05). In rats bearing the subcutaneous implants, no obvious inflammation was observed in the local tissue. CONCLUSION Poprous silicone orbital implants with uniform pores can be prepared using embedded 3D printing technology, and surface modification obviously improves hydrophilicity and biocompatibility of the silicone implants for potential clinical application.
Animals ; Rats ; Swine ; Silicon ; Orbital Implants ; Endothelial Cells ; Porosity ; Silicones ; Printing, Three-Dimensional