Roles of adipose-derived stem cells and derived exosomes in therapeutic applications to testicular injury caused by cisplatin.

In recent years, adipose-derived stem cells (ADSCs) and derived exosomes (ADSC-Ex) have been investigated for their therapeutic potential in various diseases due to their satisfactory differentiation and regeneration ability. We aimed to explore the potential treatment of ADSCs and ADSC-Ex for testicular injury caused by cisplatin. ADSCs and ADSC-Ex s were identified and extracted to treat the rat model with testicular injury caused by cisplatin. Then the immunohistochemistry and Enzyme linked immunosorbent assay (ELISA) were used to detect the potential treatment of ADSCs and ADSC-Ex. We found that ADSCs and ADSC-Ex significantly improved the testicular tissue damage, increased the number of germ cells, and improved the arrangement of the seminiferous tubules. The levels of malondialdehyde and testosterone were also improved. We speculated that ADSCs and ADSC-Ex may alleviate the testicular injury caused by cisplatin.

Single-cell transcriptome analysis reveals dynamic changes of the preclinical A549 cancer models, and the mechanism of dacomitinib.

The in vitro A549 cells, and A549 xenografts in nude mouse, were two commonly used models for anti-cancer drug discovery. However, the biological and molecular characteristics of these two classic models, and also the dynamic transcriptome changes after dacomitinib exposure remains elusive. We performed single-cell RNA sequencing to define the transcriptome profile at single-cell resolution, and processed tumor samples for bulk RNA and protein analysis to validate the differently expressed genes. Transcriptome profiling revealed that the in vitro A549 cells are heterogeneous. The minimal subpopulation of the in vitro A549 cells, which were characterized by the signature of response to unfolded protein, became the overriding subpopulation of the xenografts. The EGFR non-activating A549 cells were resistant to dacomitinib in vitro, while A549 xenografts were comparatively sensitive as EGFR-activating HCC827 xenografts. Dacomitinib inhibited MAPK signaling pathway, and increased the immune response in the A549 xenografts. A phagocytosis checkpoint stanniocalcin-1 (STC1) was significantly inhibited in dacomitinib-treated xenografts. So here our study gives the first insight of the heterogeneity of the two classic models, and the translational potential of dacomitinib being used into a broader patient population rather than EGFR common activating mutation.

A Review of the Development and Challenges of Cell Mechanical Models.

Cell models can express a variety of cell information, including mechanical properties, electrical properties, and chemical properties. Through the analysis of these properties, we can fully understand the physiological state of cells. As such, cell modeling has gradually become a topic of great interest, and a number of cell models have been established over the last few decades. In this paper, the development of various cell mechanical models has been systematically reviewed. First, continuum theoretical models, which were established by ignoring cell structures, are summarized, including the cortical membrane droplet model, solid model, power series structure damping model, multiphase model, and finite element model. Next, microstructural models based on the structure and function of cells are summarized, including the tension integration model, porous solid model, hinged cable net model, porous elastic model, energy dissipation model, and muscle model. What's more, from multiple viewpoints, the strengths and weaknesses of each cell mechanical model have been analyzed in detail. Finally, the potential challenges and applications in the development of cell mechanical models are discussed. This paper contributes to the development of different fields, such as biological cytology, drug therapy, and bio-syncretic robots.

[Application analysis of laparoscopic assisted microsurgical vasovasostomy in the treatment of vas deferens obstruction caused by inguinal herniorrhaphy].

OBJECTIVE: To investigate the effiicacy of laparoscopic assisted microsurgical vasovasostomy in the treatment of vas deferens obstruction caused by inguinal herniorrhaphy. METHODS: Clinical data of patients undergoing surgical treatment for deferential obstruction after inguinal hernia repair in the andrology department of the First Affiliated Hospital of Zhengzhou University from 2018 to 2022 were retrospectively analyzed, and they were divided into two groups according to different surgical methods: double mirror combined group and microscope group. The basic clinical data, intraoperative conditions, postoperative effects and complications of the two groups were compared. RESULTS: There were 14 cases in the double mirror group and 34 cases in the microscope group. There was no significant difference in age and history of groin operation between the two groups (P>0.05). The average length of hospital stay in the two-lens group was less than that in the microscope group (5.07±0.26 days vs 7.09±1.86 days, P< 0.01), and the average operation time in the two-lens group was more than that in the microscope group (211.93±58.55min vs 162.26±40.70min, P<0.01). The postoperative recurrence rate (85.7% vs 73.5%, P > 0.05) was similar between the two groups. There was no significant difference in early postoperative complications (0% vs 2.9%, P > 0.05). Only 1 patient in the microscope group experienced fat liquefaction and recovered after intensive dressing change. CONCLUSION: Laparoscope-assisted microscopy provides natural fertility opportunities for patients with vas deferens obstruction after inguinal hernia repair, reduces the difficulty of surgery and the length of hospital stay, and is a safe and effective surgical method comparable to traditional surgical methods.

Efficacy analysis of 26 cases of ejaculatory duct obstruction treated by prostatic utricle neck endoscopy.

Objective: To evaluate the safety and efficacy of transvesical incision in the treatment of ejaculatory duct obstruction. Methods: The clinical data of 26 male infertile patients with ejaculatory duct obstruction were retrospectively analysed at the First Affiliated Hospital of Zhengzhou University from June 2020 to August 2021. All patients were treated with seminal vesicle neck incision for ejaculatory duct obstruction. The general clinical characteristics, intraoperative conditions and postoperative effects on the patients were recorded, and the therapeutic effect was evaluated. Results: The ejaculatory duct was found through fenestration, and the seminal vesicle gland was smoothly entered in 25 patients (96.2%). Among them, 22 cases underwent bilateral endoscopy and three underwent unilateral endoscopy. Sperm appeared in 23 cases (88.5%) 3 months after surgery. The sperm concentration and motility postoperatively at 6 months were higher than that at 3 months postoperatively. No postoperative complications, such as epididymitis or retrograde ejaculation, occurred. Conclusion: Searching for the ejaculatory duct via the neck of the prostatic utricle, assisted by a low-energy holmium laser, is a new method for the treatment of ejaculatory duct obstruction. Microscopic vision is clear using this approach and the postoperative complications are few, which has high value for clinical application.

Analyzing the Clinical Efficacy of a Type 5 Phosphodiesterase Inhibitor Combined With Ziyin Baihuo Granules in the Treatment of Erectile Dysfunction.

The study explored the clinical efficacy of a type 5 phosphodiesterase inhibitor (PDE5i) combined with Ziyin Baihuo granules in the treatment of patients suffering from erectile dysfunction (ED) with yin deficiency and fire-hyperactivity syndrome. A total of 163 patients with erectile dysfunction were divided into observation and control groups. The observation group took tadalafil (Cialis) and Ziyin Baihuo granules orally, and the control group took only tadalafil orally, for 12 weeks. An additional 40 healthy people were selected as a normal group for comparison of the sex hormone levels before and after treatment of the participants in the erectile dysfunction group. After treatment, the symptoms of dry throat and tongue, tidal fever and night sweats, liking cold and avoiding heat, and waist pain showed significant improvement in the observation group (p < .05). Compared with before treatment, the clinical indexes of erectile function in the control group and the observation group were improved after treatment (p < .05). After treatment, Ziyin Baihuo granules combined with tadalafil restored the abnormal indexes of blood (p < .05) in the observation group. Our research shows that PDE5i combined with Ziyin Baihuo granules could effectively improve erectile function.

A Comparative Study on the Success Rates of Two Approaches for Seminal Vesiculoscopy.

This study aimed to compare the success rates of two approaches for seminal vesiculoscopy: through the interior of the prostatic utricle and through the neck of the prostatic utricle. The patients were divided into two groups based on the seminal vesiculoscopy used. Group A was an interior of the prostatic utricle group (152 cases), and group B was a neck of the prostatic utricle group (146 cases). The general clinical data, intraoperative conditions and surgical results of the two groups were compared. Compared with group A, group B had a higher surgical success rate (94.5% vs. 62.5%, p < .001), a shorter operation time (33 min vs. 45 min, p < .001), less blood loss (0.5 ml vs. 2 ml, p < .001), a higher pain relief rate (86.6% vs. 52.3%, p < .001), a higher remission rate of haemospermia (82.2% vs. 58.5%, p = .011), a lower recurrence rate of pain (10.4% vs. 35.4%, p < .001), a lower recurrence rate of haemospermia (15.6% vs. 37.7%, p = .014), a higher symptom remission rate of the lower urinary tract (90.9% vs. 50.0%, p = .030), a higher remission rate of scrotal moisture (84.6% vs. 45.5%, p = .042) and a higher remission rate of frequent spermatorrhea (80.0% vs. 55.6%, p = .033). Seminal vesiculoscopy undertaken through the neck of the prostatic utricle has the characteristics of high success rate, short operation time and good surgical effect and is worthy of promotion and application.

[Blood biochemical indexes in ED patients with kidney deficiency or non-kidney deficiency: A comparative analysis of 156 cases].

OBJECTIVE: To analyze the blood biochemical characteristics of the ED patients with different types of kidney deficiency or non-kidney deficiency. METHODS: We reviewed the clinical data on 156 ED patients treated in our Department of Andrology from May to July 2018 and, according to the traditional Chinese medicine (TCM) syndromes, divided them into four groups: kidney-yang deficiency (n = 48), kidney-yin deficiency (n = 34), kidney-yin+yang deficiency (n = 36) and non-kidney deficiency control (n = 38). We obtained and compared their blood biochemical indexes, including the levels of testosterone (T), estradiol (E2), cortisol (CORT), thyroid stimulating hormone (TSH), free thyroxine (FT4), free triiodothyronine (FT3), nitric oxide (NO), total nitric oxide synthase (tNOS), and inducible nitric oxide synthase (iNOS). RESULTS: There were no statistically significant differences in the mean age, course of disease, IIEF-5 score and erection hardness score (EHS) among the four groups of patients. Pairwise comparison showed that, compared with the non-kidney deficiency controls, the patients in the kidney-yin deficiency group exhibited a dramatically higher level of CORT (ï¼»87.97 ± 45.59ï¼½ vs ï¼»121.78 ± 41.87ï¼½ µg/L, P = 0.002) and those in the kidney-yang deficiency group a remarkably lower level of FT3 (ï¼»5.44 ± 0.38ï¼½ vs ï¼»5.11 ± 0.54ï¼½ pmol/L, P = 0.008). The iNOS level was significantly higher in the kidney-yin deficiency group (14.42 ± 2.49 U/ml) than in either the control (12.71 ± 2.58 U/ml) (P = 0.039) or the kidney-yang deficiency group (13.05 ± 2.17 U/ml) (P =0.049). CONCLUSIONS: ED patients with different types of kidney deficiency syndromes have different blood biochemical indexes, which may help clarify the biological basis of the TCM syndromes of kidney deficiency in ED patients.

[Factors influencing recovery from childhood inguinal herniorrhaphy-induced vas deferens obstruction after microscopic vasovasostomy].

OBJECTIVE: To investigate the factors influencing the recovery from childhood inguinal herniorrhaphy (IH)-induced vas deferens obstruction following microscopic vasovasostomy. METHODS: We retrospectively analyzed the clinical data on 41 cases of microscopic vasovasostomy for obstructive azoospermia in our hospital from July 2015 to September 2018. All the patients had a history of inguinal hernia treated by IH in the childhood. We performed scrotal ultrasonography, semen analysis and seminal plasma biochemistry to confirm vas deferens obstruction preoperatively. If sperm was observed for ≥2 times in semen examination after vasovasostomy, we considered the vas deferens successfully unobstructed. RESULTS: Microscopic vasovasostomy was successfully completed in 39 of the cases, of which2 were lost to follow-up, with a follow-up rate of 94.8% (37/39). The patients, at the mean age of (25.54 ± 2.85) years and with body mass index (BMI) of (24.92 ± 2.79) kg/m2 and post-IH time of (18.97 ± 2.58) years, were followed up for (13.05 ± 3.74) months. Successful recovery from vas deferens obstruction was observed in 78.4% (29/37) of the patients after IH, 80.0% (16/20) in the < 26-year-olds, 76.5% (13/17) in the ≥26-year-olds (P = 0.795), 75.0% (12/16) in those with BMI < 24.92 kg/m2 , 81.0% (17/21) in those with BMI ≥24.92 kg/m2 (P = 0.807), 78.6% (11/14) in those with post-IH time of < 19 years, 18.3% (18/23) in those with post-IH time of ≥19 years (P = 0.982), 60.0% (12/20) in those with sperm and 82.4% (14/17) in those without sperm found intraoperatively (P = 0.428), 42.9% (3/7) in those treated by unilateral and 82.4% (26/30) in those by bilateral vasovasostomy (P = 0.027). Multivariate logistic regression analysis showed a close correlation between the operation side and postoperative recovery from vas deferens obstruction (P = 0.022). CONCLUSIONS: For male patients undergoing microscopic vasovasostomy for childhood IH-induced vas deferens obstruction, the operation side is an independent factor influencing postoperative recovery, while age, BMI, post-IH time, and intraoperative presence or absence of sperm are not significantly correlated with it.

MLPH Accelerates the Epithelial-Mesenchymal Transition in Prostate Cancer.

INTRODUCTION: Prostate cancer (PC) is the second greatest cause of cancer deaths globally. PC presents a poor prognosis once it metastasizes. There is considerable proof of vital epithelial-mesenchymal transition (EMT) functionality in PC metastasis. Previous studies revealed that melanophilin (MLPH) is associated with PC; however, its role in PC remains poorly understood. METHODS: Bioinformatics analyses were performed. The cellular responses to MLPH knockdown were examined in HCC cell lines via wound healing assay, migration and invasion assay, Western blotting. RESULTS: Analysis of the PROGgeneV2 database revealed that high MLPH expression might indicate poor overall survival. MLPH knockdown reduced PC cell migration, proliferation, and invasion. MLPH downregulation in vivo resulted in a lower growth rate and fewer metastatic nodules in lung tissues. Furthermore, MLPH knockdown recovered downregulated expression of the mesenchymal marker N-cadherin and the epithelial marker E-cadherin following a decrease in ß-catenin. CONCLUSION: These results indicate that progression of PC is stimulated via MLPH-dependent initiation of the EMT.

Icariside II facilitates the differentiation of ADSCs to schwann cells and restores erectile dysfunction through regulation of miR-33/GDNF axis.

BACKGROUND: Adipose derived stem cells (ADSCs) have the property to differentiate into neuron-like cells, which may provide a novel insight for the restoration of erectile dysfunction (ED) mainly induced by cavernous nerve injury. Icariside II (ICA II) has been reported to play a key role in the regulation of erectile function via stimulating the differentiation of ADSCs to Schwann Cells (SCs). However, the function and molecular mechanisms of ICA II in ED remains to be further clarified. METHODS: The expression of S100, P75, GDNF and miR-33 was detected by qRT-PCR. And the relative proteins expression was determined by western blot. Cell viability was measured by Cell Counting Kit-8 (CCK-8) assay. Bioinformatics, luciferase reporter and RNA immunoprecipitation (RIP) assays were performed to verify the interaction between miR-33 and GDNF. Intracavernosal pressure (ICP), the ratio of ICP and mean arterial pressure (MAP), as well as nNOS expression were examined to evaluate the erectile function of SD rats with bilateral cavernous nerve injury (BCNI). RESULTS: ICA II and miR-33 respectively promoted and inhibited the differentiation of ADSCs to SCs. MiR-33 could negatively regulate P75 and GDNF expression. ICA II exerted promotion effects on differentiation of ADSCs to SCs via regulating miR-33. GDNF was identified to be a target of miR-33. MiR-33 overexpression abrogated the stimulatory effect of ICA II on ADSCs' differentiation, which was blocked by GDNF overexpression. treated with ICA II recovered the erectile function of BCNI model rats through regulation of miR-33. CONCLUSION: ICA II contributed to the differentiation of ADSCs to SCs viamiR-33/GDNF axis, contributing to the recovery of erectile function in BCNI rats.

[Efficacy and safety of regular use of sildenafil in the treatment of penile erectile dysfunction].

OBJECTIVE: To investigate the efficacy and safety of regular oral use of sildenafil in the treatment of ED. METHODS: We randomly divided 334 ED patients into three groups to be treated orally with sildenafil tablets at 50 mg qd (sildenafil regular), sildenafil tablets at 100 mg 30 minutes before intercourse (sildenafil on-demand), and tadalafil tablets at 10 mg qd (tadalafil regular), all for 3 months. Then we recorded the IIEF-5 score and penile erection hardness score (EHS) and adverse reactions and compared them among the three groups of patients. RESULTS: There were no statistically significant differences among the three groups of patients in age, body mass index, education, ED duration, or baseline IIEF-5 and EHS (P > 0.05). After 3-month medication, both IIEF-5 score and EHS were significantly improved in the three groups of patients as compared with the baseline (P < 0.05), with no statistically significant difference in the IIEF-5 score among the sildenafil regular, sildenafil on-demand and tadalafil regular groups (15.15 ± 2.05 vs 15.55 ± 2.36 vs 15.54 ± 2.27, P > 0.05), but the EHS markedly higher in the sildenafil on-demand than in the sildenafil regular group (3.48 ± 1.80 vs 3.12 ± 1.52, P < 0.05). The effectiveness rates in the sildenafil regular, sildenafil on-demand and tadalafil regular groups were 76.2%, 62.4% and 80.8%, respectively, significantly lower in the sildenafil on-demand than in the other two groups (P < 0.05). Adverse reactions were mild and showed no statistically significant difference in the incidence rate among the three groups (P > 0.05). CONCLUSIONS: Regular use of sildenafil has a therapeutic effect similar to that of tadalafil but better than that of sildenafil on-demand, without more adverse effects.

Icariside II Promotes the Differentiation of Adipose Tissue-Derived Stem Cells to Schwann Cells to Preserve Erectile Function after Cavernous Nerve Injury.

Icariside II (ICA II) is used in erectile dysfunction treatment. Adipose tissue-derived stem cells (ADSCs) are efficient at improving erectile function. This study aimed to explore the action mechanism of ADSCs in improving erectile function. ADSCs were isolated from the adipose tissues of rats. Cell proliferation was determined using the Cell Counting Kit-8 (CCK-8) assay. The expressions of mRNA and protein were determined separately through qRT-PCR and western blot. The endogenous expressions of related genes were regulated using recombinant plasmids and cell transfection. A Dual-Luciferase Reporter Assay was performed to determine the interaction between miR-34a and STAT3. Rat models with bilateral cavernous nerve injuries (BCNIs) were used to assess erectile function through the detection of mean arterial pressure (MAP) and intracavernosal pressure (ICP). ICA II promoted ADSCs' proliferation and differentiation to Schwann cells (SCs) through the inhibition of miR-34a. Suppressed miR-34a promoted the differentiation of ADSCs to SCs by upregulating STAT3. ICA II promoted the differentiation of ADSCs to SCs through the miR-34a/STAT3 pathway. The combination of ICA II and ADSCs preserved the erectile function of the BCNI model rats. ADSCs treated with ICA II markedly preserved the erectile function of the BCNI model rats, which was reversed through miR-34a overexpression. ICA II promotes the differentiation of ADSCs to SCs through the miR-34a/STAT3 pathway, contributing to erectile function preservation after the occurrence of a cavernous nerve injury.

Genipin inhibits the growth of human bladder cancer cells via inactivation of PI3K/Akt signaling.

Genipin, a natural compound derived from the fruit of Gardenia jasminoides, possesses numerous biological properties. The aim of the present study was to investigate the anticancer effects of genipin in human bladder cancer. T24 and 5637 bladder cancer cells were treated with different concentrations of genipin (0-200 µM) and tested for cell viability, colony formation, cell cycle progression and apoptosis. A xenograft model of bladder cancer was established to determine the anticancer effect of genipin in vivo. The involvement of the phosphoinositide-3 kinase (PI3K)/Akt pathway in the action of genipin was examined. Genipin treatment significantly inhibited the viability and clonogenic growth of bladder cancer cells and inhibited the growth of T24 xenograft tumors, compared with vehicle controls (P<0.05). Genipin-treated cells exhibited a cell cycle arrest at the G0/G1-phase, which was accompanied by a deregulation of numerous cell cycle regulators. Genipin-treated cells demonstrated a significant increase in the percentage of apoptotic cells, loss of mitochondrial membrane potential, Bax translocation to the mitochondria and the release of cytochrome c to the cytosol. Additionally, genipin treatment significantly (P<0.05) reduced the phosphorylation levels of PI3K and Akt in bladder cancer cells. Importantly, genipin-mediated anticancer effects were reversed by the overexpression of constitutively active Akt. In conclusion, to the best of our knowledge, the present study demonstrates for the first time the growth inhibitory effects of genipin in bladder cancer cells, and indicates its potential as a natural anticancer agent for bladder cancer.

Aroclor1254 disrupts the blood-testis barrier by promoting endocytosis and degradation of junction proteins via p38 MAPK pathway.

The blood-testis barrier (BTB) constituted by coexisting junction apparatus between Sertoli cells (SCs) plays an important role in spermatogenesis, which is a known target of various environmental toxicants. The commercial polychlorinated biphenyls mixture, Aroclor1254, has been shown to impair male reproduction by decreasing sperm count and affecting SC metabolism. This study was designed to investigate the effects of Aroclor1254 on the BTB integrity and elucidate the underlying mechanisms. We found that Aroclor1254 treatment in rats (1 or 3 mg/kg per day for 21 consecutive days) and in primary cultured SCs (5 or 10 µg/ml for 48 h) could induce BTB disruption via p38 MAPK pathway, concurrently with increments in junction proteins (JAM-A, N-cadherin, and ß-catenin) endocytosis, and occludin ubiquitination. Either inhibition of caveolin-dependent membrane protein internalization by cholesterol oxidase or silencing E3 ubiquitine ligase Itch by small interfering RNA could partially counteract the effects of Aroclor1254 on the barrier function of cultured SCs. These results demonstrate that Aroclor1254 disrupts the BTB function by promoting the caveolin-dependent endocytosis and ubiquitine-proteasome degradation of junction proteins through the p38 MAPK pathway, which might be the potential reasons for its negative effects on spermatogenesis and male reproduction.

Low concentration of BPA induces mice spermatocytes apoptosis via GPR30.

Bisphenol A (BPA) acts as xenoestrogen and has a great impact on disorders of human reproductive system. However, the mechanism through which BPA can affect human testicular function remains to be identified. GPR30 is a novel membrane estrogen receptor with high-affinity and low-capacity binding to estrogens. We demonstrated that estrogen receptor α (ERα), estrogen receptor ß (ERß) as well as GPR30 are expressed in mouse spermatocyte-derived GC-2 cells using Real-time PCR. We treated the cells with different doses of BPA and found that even low doses of BPA can inhibit GC-2 cell growth using MTT assay. To make sure which receptor is responsible for the biological function of BPA, we used ER down-regulator ICI and indicated that BPA could bind to GPR30. We also observed that BPA was able to induce Erk1/2 phosphorylation in GC-2 cells and proved that this process was mediated by GPR30-related EGFR-MAPK pathway using western blot. By Real-time PCR, we found that the expression of c-Fos was up-regulated and Cyclin D1 gene was down-regulated, in the presence of BPA and ICI. The results of MTT assay, comet assay and flow cytometry indicated that the activation of GPR30 induced by BPA inhibited the cell growth and induced cell apoptosis and ICI, GPR30 siRNA, EGFR inhibitor (AG), and MAPK (PD) inhibitor could partially reverse this effect. Immunohistochemistry on the testis of BPA -damaged mice showed that BPA induced spermatocyte apoptosis without affecting the seminiferous tubules and spermatocyte. In conclusion, BPA triggered spermatocyte apoptosis via GPR30.

A novel method to calculate the mechanical properties of cancer cells based on atomic force microscopy.

PURPOSE: Mechanical properties, as the inherent characteristics of cells, play a critical role in many essential physiological processes, including cell differentiation, migration, and growth. The mechanical properties of cells are one of the criteria that help to determine whether the tissue contains lesions at the single cell level, and it is very important for the early prevention and accurate diagnosis of diseases. Atomic force microscopy (AFM) makes it possible to measure the mechanical properties at single cell level in physiological state. This paper presents a novel method to calculate the mechanical properties of cancer cells more accurately through Atomic force microscopy. METHODS: A new induced equation of Hertz's model, called differential Hertz's model, has been proposed to calculate the mechanical properties of cancer cells. Moreover, the substrate effect has also been effectively reduced through comparing the calculated mechanical properties of cell at different cell surface areas. RESULTS: The results indicate that the method utilized to calculate the mechanical properties of cells can effectively eliminate the errors in calculation, caused by the thermal drift of AFM system and the substrate effect, and thus improve the calculation accuracy. CONCLUSION: The mechanical properties calculated by our method in this study are closer to the actual value. Thus, this method shows potential for use in establishing a standard library of Young's modulus.

Synthetic Bax-Anti Bcl2 combination module actuated by super artificial hTERT promoter selectively inhibits malignant phenotypes of bladder cancer.

BACKGROUND: The synthetic biology technology which enhances the specificity and efficacy of treatment is a novel try in biomedical therapy during recent years. A high frequency of somatic mutations was shown in the human telomerase reverse transcriptase (hTERT) promoter in bladder cancer, indicating that a mutational hTERT promoter might be a tumor-specific element for bladder cancer therapy. In our study, we aimed to construct a synthetic combination module driven by a super artificial hTERT promoter and to investigate its influence on the malignant phenotypes of bladder cancer. METHODS: The dual luciferase assay system was used to verify the driven efficiency and tumor-specificity of the artificial hTERT promoter and to confirm the relationship between ETS-1 and the driven efficiency of the artificial hTERT promoter. CCK-8 assay and MTT assay were used to test the effects of the Bax-Anti Bcl2 combination module driven by the artificial hTERT promoter on cell proliferation. Simultaneously, the cell apoptosis was detected by the caspase 3ELISA assay and the flow cytometry analysis after transfection. The results of CCK-8 assay and MTT assay were analyzed by ANOVA. The independent samples t-test was used to analyze other data. RESULTS: We demonstrated that the artificial hTERT promoter had a higher driven efficiency which might be regulated by transcription factor ETS-1 in bladder cancer cells, compared with wild-type hTERT promoter. Meanwhile, the artificial hTERT promoter showed a strong tumor-specific effect. The cell proliferation inhibition and apoptosis induction were observed in artificial hTERT promoter- Bax-Anti Bcl2 combination module -transfected bladder cancer 5637 and T24 cells, but not in the module -transfected normal human fibroblasts. CONCLUSION: This module offers us a useful synthetic biology platform to inhibit the malignant phenotypes of bladder cancer in a more specific and effective way.

[Transurethral seminal vesiculoscopy for the treatment of vesiculitis with hemospermia:A report of 64 cases].

Objective: To explore the treatment of vesiculitis with hemospermia by transurethral seminal vesiculoscopy. Methods: We treated 64 cases of vesiculitis with hemospermia by transurethral seminal vesiculoscopy. During the operation,we removed the stones and inflammatory substances and collected seminal vesicle fluid to be cultured for bacteria,ureaplasma urealyticum(UU),chlamydia trachomatis(CT),and mycoplasma hominis(MH),followed by infusion of levofloxacin at 0. 3 g/100 ml into the seminal vesicle. Regular follow-up was conducted post-operatively. Results: All the operations were successfully accomplished, the operation time averaging(40 ± 15) min(25- 50 min). The ejaculatory duct opening was observed on the verumontanum surface in the posterior urethra in 2 cases, abnormal passages found in the prostatic utricle in 8 cases, and seminal vesicle fenestration from the prostatic utricle conducted in the other 54 cases(32 by seminal vesiculoscopy and 22 with holmium laser). Stones were seen in the prostatic utricle in 5 cases, in the seminal vesicle in 6 cases, and in both the prostatic utricle and seminal vesicle in 2 cases. Culture of the seminal vesicle fluid showed the acinetobacter to be positive in 1 case and UU, CT, and MH to be negative. At 3 months after surgery, hemospermia was cured in 52 cases, relieved in 8,and unimproved in 4. Conclusion: Seminal vesicle fenestration drainage by transurethral seminal vesiculoscopy for the treatment of vesiculitis with hemospermia has the advantages of short operation time, high effectiveness and no obvious complications and can also be employed for the examination of the seminal vesicle as well as removal of stones and inflammatory substances.

Inhibition of glucose-transporter 1 (GLUT-1) expression reversed Warburg effect in gastric cancer cell MKN45.

Glucose transporter-1 (GLUT-1) plays critical roles in cancer development and progression. Warburg effect (aerobic glycolysis) contributes greatly to tumorigenesis and could be targeted for tumor therapy. However, published data on the relationship between GLUT-1 and Warburg effect are scarce. In this study, gastric cancer cell, MKN45, was transfected with GLUT-1 shRNA using Lipofectamine 2000. Oxygen consumption, LDH activity, lactate production and cytoplasmic pyruvate were detected after MKN45 cells with GLUT-1 knockdown. In the last, hexokinase 1 (HK1), HK2, and pyruvate kinase M2 (PKM2) expression were detected by using western blot. In this study, we showed that inhibition of GLUT-1 expression reversed Warburg effect in MKN45 cells, and induced apoptosis.