In vitro study of neuroendocrine regulation over the testicular development in mud crabs Scylla serrata

The reproductive endocrine activities of neuroendocrine organs and androgenic glands (AG) in male Scylla serrata were investigated with co-incubation technology. In vitro studies show that: (1) the AG in Stage Ⅲ can significantly accelerate the development of seminiferous tubules and spermic matu- ration; (2) the brain, thoracic ganglia and optic ganglia have no direct influence over the testicular devel- opment; (3) the brain and thoracic ganglia can significantly promote the growth of the AG cells and highly significantly boost the ratio of Type B cells, while the optic ganglia have no such effects. It is the first time for in vitro investigations to confirm that the brain and thoracic ganglia can regulate testicular development through AG in male crustaceans.

A STUDY OF DECREASING DRUG-RESISTANCE OF HUMAN OVARIAN CANCER CELLS IN VITRO

A chemosensitivity test for ovarian cancer using tritiated thymidine incorporation assay was carried out. A dose-response relationship for cisplatin and potentiation of verapamil in increasing vincristine inhibition to ovarian cancer were investigated. A 5- fold increase of cisplatin density converted the tumors which were initially resistance to standard-dose cisplatin Into drug-sensitive ones. Vera-pamil was found to be able to overcome vincristine-resistance of some tumors in vitro. These results suggest that using high dose cisplatin therapy or increasing local drug concentration by using other administration way, we could expect some ovarian cancers that had failed to standard dose cisplatin therapy to be effective. Combination of vincristine with verapamll may be helpful in treating some vincristineresistant cases.

Radiation Induced Bystander Effect: From in Vitro Studies to Clinical Application

In the past 20 years, the classic paradigm in radiobiology recognizing DNA as the main target for the action of radiation has changed. The new paradigm assumes that both targeted and non-targeted effects of radiation determine the final outcome of irradiation. Radiotherapy is one of the main modality treatments of neoplastic diseases with intent to cure, or sometimes to palliate only, thus radiation-induced non-targeted effect, commonly referred to as the radiation-induced bystander effect (RIBE) may have a share in cancer treatment. RIBE is mediated by molecular signaling from radiation targeted cells to their non-irradiated neighbors, and comprises such phenomena as bystander effect, genomic instability, adaptive response and abscopal effect. Whereas first three phenomena may appear both in vitro and in vivo, an abscopal effect is closely related to partial body irradiation and is a systemic effect mediated by immunologic system which synergizes with radiotherapy. From the clinical point of view abscopal effect is particularly interesting due to both its possible valuable contribution to the treatment of metastases, and the potential harmful effects as induction of genetic instability and carcinogenesis. This review summarized the main results of investigations of non-targeted effects coming from in vitro monolayer cultures, 3-dimentional models of tissues, preclinical studies on rodents and clinically observed beneficial abscopal effects with particular emphasis on participation of immunotherapy in the creation of abscopal effects.

In Vitro Chemosensitivity Testing of Primary and Recurrent Breast Carcinomas and Its Clinical Significance

In this study, in vitro chemosensitivity testing was conducted on primary cultured breast cancer cells from 96 patients with breast cancer, and the results showed that the cells from a few patients with primary breast cancer developed multidrug resistance （MDR） prior to the first chemotherapy exposure. All the cells from the recurrent cancer patients had MDR. The findings suggested that patients having MDR would benefit from high-dose chemotherapy （HDC） regimens. In vitro chemo-sensitivity screening, which was aimed at improving the therapeutic efficacy and minimizing side effects, helps in choosing individualized treatment for breast cancer.

In Vitro Studies: Potensial Use of Dyospiros kaky, Avium grafeolens, Guazuma ulmifolia, Hibiscus sabdariffa as an Agent of Anti-cholesterol

God has made man and has prepared everything for human life.This universe was created for humans to be healthy.The lifestyle of humans in the millenial era has made circulatory disorders in the form of hypercholesterolemia.This study aims to determine the efficacy of herbal plants to reduce blood cholesterol levels.The method used is the in vitro method.Herbal plant used in this experiment Dyospiros kaky 50%,Avium grafeolens 20%,Guazuma ulmifolia 20%,Hibiscus sabdariffa 10%.This study showed that cholesterol level decreased from 200 mg/dL to 102 mg/100 g at dose 20 mg/dL,64.5 mg/100 g,at dose 40 mg/dL,and 94.2 mg/100 g,at dose 60 mg/dL.The best dose to decrease blood cholesterol was 40 mg/dL.This study indicated that Dyospiros kaki is an agent of anti-cholesterol.

Experimental study of TGF-β1/Smads pathway inhibition of macrophage polarization based on miR145-5P negative feedback regulation

imbalance of synovial macrophages in patients with rheumatoid arthritis(RA).Methods:Human mononuclear cells(THP‑1)at logarithmic growth stage were induced into M1‑type macrophages,and RA synovial fibroblasts M1‑type macrophages were co‑cultured into synovial macrophages.Synovial macrophages were divided into four groups:RA group(blank group),TGF‑β1 group(model group)and miR145‑5P overexpression group(TGF‑β1+miR145‑5P mimics group)and miR145‑5P overexpression negative control group(TGF‑β1+miR145‑5P‑mimics‑NC group).The blank group did not receive any treatment,and the other three groups were induced by TGF‑β1 in the medium for 48 h.Transfection miR145‑5p mimic and miR145‑5P‑mimics‑NC were added to co‑culture medium,and IL‑6,IL‑6 and IL‑6 of synovial macrophages were detected by ELISA.CD163 expression.Rt‑qpcr was used to detect miR145‑5p mRNA,TGF‑β1mRNA,Smad3mRNA,Smad7mRNA expression level.The expression of TGF‑β1/Smads pathway related proteins was detected by Western Blotting.Results:Compared with blank group,IL‑6 level was up‑regulated(P<0.01),and CD163 level was down‑regulated in model group(P<0.05),suggesting that TGF‑β1 could induce intensified immune inflammatory response.Compared with the negative miR145‑5P overexpression control group and model group,The expression of miR145‑5P overexpression group molecule CD163 was significantly increased by ELISA(P<0.01),and the expression of inflammatory factor IL‑6 was decreased(P<0.05).PCR showed that miR145‑5P mRNA expression level was significantly increased in miR145‑5P overexpression group,Smad3 mRNA and TGF‑β1 mRNA were significantly decreased,and Smad7 mRNA was significantly increased(P<0.01).WB method showed that the anti‑inflammatory protein Smad7 was significantly increased,while TGF‑β1 and Smad3 were significantly decreased(P<0.01).Transwell chamber results confirmed that miR145‑5P overexpression group significantly reduced macrophage invasion(P<0.01).Correlation analysis showed that miR145‑5P was negatively correlated with Smad3 and positively correlated with Smad7(P<0.01).Conclusion:miR145‑5P may inhibit macrophage polarization in RA patients by targeting Smad3 protein,negatively regulating TGF‑β1/Smads pathway,and alleviating immune inflammation.

Understanding the Recovery of the Intervertebral Disc:A Comprehensive Review of In Vivo and In Vitro Studies

Within the consistent daily rhythm of human life,intervertebral discs endure a variety of complex loads beyond the influences of gravity and muscle forces,leading to significant morphological changes(in terms of volume,area,and height)as well as biomechanical alterations,including an increase in disc stiffness and a decrease in intradiscal pressure.Remarkably,the discs demonstrate an ability to regain their original morphological and biomechanical characteristics after a period of nocturnal rest.The preservation of normal disc function is critically dependent on this recovery phase,which serves to forestall premature disc degeneration.This phenomenon of disc recovery has been extensively documented through numerous in vivo studies employing advanced clinical techniques such as Magnetic Resonance Imaging(MRI),stadiometry,and intradiscal pressure measurement.However,the findings from in vitro studies present a more complex picture,with reports varying between full recovery and only partial recuperation of the disc properties.Moreover,research focusing on degenerated discs in vitro has shed light on the quantifiable impact of degeneration on the disc ability to recover.Fluid dynamics within the disc are considered a primary factor in recovery,yet the disc intricate multiscale structure and its viscoelastic properties also play key roles.These elements interact in complex ways to influence the recovery mechanism,particularly in relation to the overall health of the disc.The objective of this review is to collate,analyze,and critically evaluate the existing body of in vivo and in vitro research on this topic,providing a comprehensive understanding of disc recovery processes.Such understanding offers a blueprint for future advancements in medical treatments and bionic engineering solutions designed to mimic,support,and enhance the natural recovery processes of intervertebral discs.

The Inhibitory Effect of Oridonin on the Growth of Fifteen Human Cancer Cell Lines

OBJECTIVE To study the inhibitory effect of oridonin on the growth of cancer cells. METHODS Fifteen human cancer cell lines were subjected to various concentrations of oridonin in culture medium. The inhibitory rate of cell growth was measured by the MTT assay, and compared with a negative control and 5-Fu-positive control. RESULTS The 50% inhibiting concentration （IC50） and maximal inhibition （Imax） of oridonin shown by studying the growth of the cancer cell lines were as follows： leukemias （HL60 cells： 3.9 μg/ml and 73.8%, K562 cells： 4.3 μg/ml and 76.2%）; esophageal cancers（SHEEC cells： 15.4 μg/ml and 99.2%, Eca109 cells： 15.1 μg/ml and 84.6%, TEl cells： 4.0 μg/ml and 70.2%）; gastric cancers （BGC823 cells： 7.6 μg/ml and 98.7%, SGC7901 cells： 12.3 μg/ml and 85.7%）; colon cancers （HT29 cells： 13.6 μg/ml and 97.2%, HCT cells： 14.5 μg/ml and 96.5%）; liver cancers （Bel7402 cells： 15.2 μg/ml and 89.2%, HepG2 cells： 7.1 μg/ml and 88.3%）; pancreatic cancer （PC3 cells： 11.3 μg/ml and 68.4%）; lung cancer （A549 cells： 18.6 μg/ml and 98.0% ）; breast cancer （MCF7 cells： 18.4 μg/ml and 84.7%）; uterine cervix cancer （Hela cells： 13.7 μg/ml and 98.5%）. CONCLUSION Oridonin had a relatively wide anti-tumor spectrum, and a relatively strong inhibitory effect on the growth of the 15 human cancer cells. Inhibitory effects were concentration dependent.

Preparation and evaluation of sustained-release diltiazem hydrochloride pellets

In this study,diltiazem hydrochloride(DTZ)pellets were prepared successfully by extrusionespheronization method.Then methacrylic acid and ethylcellulose coating formulations were employed to make the DTZ pellets sustained release.The pellets with different coatings were investigated by in vitro dissolution tests.At last,the pellets with the best coating copolymer were subjected to pharmacokinetic studies in beagle dogs.The dissolution profiles of pellets coated with Eudragit
NE30D were similar to Herbesser
,one of the marketed sustained release capsules.In the bioavailability study,the principal pharmacokinetic parameters of self-made pellets and the marketed ones were comparable;the relative bioavailability of DTZ sustained release capsules compared with Herbesser
was 98.5
36.4%.All the data indicated self-made sustained pellets could prolong the release of DTZ,decrease the fluctuation of drug level in vivo,and increase the compliance of patients.

Dynamic changes in proprotein convertase 2 activity in cortical neurons after ischemia/reperfusion and oxygen-glucose deprivation

In this study, a rat model of transient focal cerebral ischemia was established by performing 100 minutes of middle cerebral artery occlusion, and an in vitro model of experimental oxygen-glucose deprivation using cultured rat cortical neurons was established. Proprotein convertase 2 activity gradually decreased in the ischemic cortex with increasing duration of reperfusion. In cultured rat cortical neurons, the number of terminal deoxynucleotidyl transferase-mediated 2＇-deoxyuridine 5＇-triphosphate-biotin nick end labeling-positive neurons significantly increased and proprotein convertase 2 activity also decreased gradually with increasing duration of oxygen-glucose deprivation. These experimental findings indicate that proprotein convertase 2 activity decreases in ischemic rat cortex after reperfusion, as well as in cultured rat cortical neurons after oxygen-glucose deprivation. These changes in enzyme activity may play an important pathological role in brain injury.

Antimicrobial activity of topical agents against Propionibacterium acnes： an in vitro study of clinical isolates from a hospital in Shanghai, China

This study aimed to compare the antimicrobial activities of topical agents against Propionibacterium acnes isolated from patients admitted to a hospital in Shanghai, China. The minimal inhibitory concentrations of the cultured P. acnes were determined in accordance with the Clinical and Laboratory Standards Institute. Susceptibilities to clindamycin and erythromycin were compared in terms of gender, age, disease duration, previous treatment, and disease severity. A total of 69 P. acnes strains were isolated from 98 patients （70.41%）. The susceptibility to triple antibiotic ointment （neomycin/bacitracin/polymyxin B） and bacitracin was 100%. The susceptibility to fusidic acid was 92.7%. The resistance rates to neomycin sulfate, erythromycin, and clindamycin were 11.7%, 49.3%, and 33.4%, respectively. The high resistance rate to clindamycin and erythromycin was significantly affected by gender, previous treatment, and disease severity rather than by age and disease duration. Topical antibiotics should not be used separately for long-term therapy to avoid multiresistance. The use of topical antibiotics should be determined by clinicians on the basis of clinical conditions.

EFFECTS OF PARATHYROID HORMONE AND ESTRADIOL ON PROLIFERATION AND FUNCTION OF HUMAN OSTEOBLASTS FROM FETAL LONG BONE AN IN VITRO STUDY

The effect of parathyroid hormone (PTH) (0.01 nM-10 nM) and 17 -estradiol (E2, 1 nmol-10 nM) alone or in combination on 3H thymidine incorporation, alkaline phosphatase and adenylate cyclase activities were investigated in human fetal osteoblasts using serum-free monolayer primary cultures. The results showed that PTH inhibited cell proliferation while E, promoted it. On alkaline phosphatase activity, PTH showed a complex results while E, were slightly inhibitory. PHT-E2 combination suggested that E2 could alter the effect of PTH alone, also potentiated the anabolic and antagonize the catabolic effects of PTH on bone formation.

Chinese Medicine for Treating Diabetic Nephropathy

Diabetic nephropathy is one of the main causes of renal end-stage disease. The pathogenesis of diabetic nephropathy is complex. The current treatment is only for a particular cause without multi-target therapeutic drugs. Chinese medicine is a great treasure with multi-component complex drugs interacting with multiple targets and functions. This paper reviewed the protective effect of Chinese medicine for treating diabetic nephropathy in clinical studies, in vivo studies, and in vitro studies. The possible mechanisms, the major compounds and active crude drugs were also summarized. It was shown that Chinese medicine could not only relieve several symptoms and improve the quality of life, but also reduce the levels of proteinuria and kidney damage, and further improve renal function via multiple pathways based on the whole human system. Moreover, there were no reports of severe adverse reactions during the treatment.