Targeted optimization of central carbon metabolism for engineering succinate production in Escherichia coli.

BACKGROUND: Succinate is a kind of industrially important C4 platform chemical for synthesis of high value added products. Due to the economical and environmental advantages, considerable efforts on metabolic engineering and synthetic biology have been invested for bio-based production of succinate. Precursor phosphoenolpyruvate (PEP) is consumed for transport and phosphorylation of glucose, and large amounts of byproducts are produced, which are the crucial obstacles preventing the improvement of succinate production. In this study, instead of deleting genes involved in the formation of lactate, acetate and formate, we optimized the central carbon metabolism by targeting at metabolic node PEP to improve succinate production and decrease accumulation of byproducts in engineered E. coli. RESULTS: By deleting ptsG, ppc, pykA, maeA and maeB, we constructed the initial succinate-producing strain to achieve succinate yield of 0.22 mol/mol glucose, which was 2.1-fold higher than that of the parent strain. Then, by targeting at both reductive TCA arm and PEP carboxylation, we deleted sdh and co-overexpressed pck and ecaA, which led to a significant improvement in succinate yield of 1.13 mol/mol glucose. After fine-tuning of pykF expression by anti-pykF sRNA, yields of lactate and acetate were decreased by 43.48 and 38.09 %, respectively. The anaerobic stoichiometric model on metabolic network showed that the carbon fraction to succinate of engineered strains was significantly increased at the expense of decreased fluxes to lactate and acetate. In batch fermentation, the optimized strain BKS15 produced succinate with specific productivity of 5.89 mmol gDCW(-1) h(-1). CONCLUSIONS: This report successfully optimizes succinate production by targeting at PEP of the central carbon metabolism. Co-overexpressing pck-ecaA, deleting sdh and finely tuning pykF expression are efficient strategies for improving succinate production and minimizing accumulation of lactate and acetate in metabolically engineered E. coli.

Esophageal squamous cell carcinoma metastases to kidney and renal hilar lymph nodes through epithelial-mesenchymal transition: a case report and literature review.

BACKGROUND: Esophageal cancer (EC) metastasized to the kidney is extremely rare clinically. Here, we present a case of metachronous renal metastasis of esophageal squamous cell carcinoma (ESCC) through epithelial-mesenchymal transition (EMT). CASE PRESENTATION: A 60-year-old patient, male, complained of left waist pain for 5 days, 11 months after radical esophagectomy. Laboratory tests revealed haematuria. Both CT and PET-CT scan showed retroperitoneal lymph nodes and left renal masses. Subsequently the patient received a left nephrectomy and lymph nodes resection, and squamous cell carcinoma of kidney and renal hilar lymph nodes was diagnosed, combined with morphology, medical history and immunophenotype, it was presumed to be metastasis of ESCC through the EMT pathway. CONCLUSIONS: The renal metastasis of squamous cell carcinoma should be considered in patients with history of EC, although this is very rare. Histopathological examination combined with immunochemical detection is helpful in differential diagnosis.

Throat microbiota drives alterations in pulmonary alveolar microbiota in patients with septic ARDS.

OBJECTIVES: The translocation of intestinal flora has been linked to the colonization of diverse and heavy lower respiratory flora in patients with septic ARDS, and is considered a critical prognostic factor for patients. METHODS: On the first and third days of ICU admission, BALF, throat swab, and anal swab were collected, resulting in a total of 288 samples. These samples were analyzed using 16S rRNA analysis and the traceability analysis of new generation technology. RESULTS: On the first day, among the top five microbiota species in abundance, four species were found to be identical in BALF and throat samples. Similarly, on the third day, three microbiota species were found to be identical in abundance in both BALF and throat samples. On the first day, 85.16% of microorganisms originated from the throat, 5.79% from the intestines, and 9.05% were unknown. On the third day, 83.52% of microorganisms came from the throat, 4.67% from the intestines, and 11.81% were unknown. Additionally, when regrouping the 46 patients, the results revealed a significant predominance of throat microorganisms in BALF on both the first and third day. Furthermore, as the disease progressed, the proportion of intestinal flora in BALF increased in patients with enterogenic ARDS. CONCLUSIONS: In patients with septic ARDS, the main source of lung microbiota is primarily from the throat. Furthermore, the dynamic trend of the microbiota on the first and third day is essentially consistent.It is important to note that the origin of the intestinal flora does not exclude the possibility of its origin from the throat.

Metabolic engineering of Escherichia coli for production of salvianic acid A via an artificial biosynthetic pathway.

Salvianic acid A, a valuable derivative from L-tyrosine biosynthetic pathway of the herbal plant Salvia miltiorrhiza, is well known for its antioxidant activities and efficacious therapeutic potential on cardiovascular diseases. Salvianic acid A was traditionally isolated from plant root or synthesized by chemical methods, both of which had low efficiency. Herein, we developed an unprecedented artificial biosynthetic pathway of salvianic acid A in E. coli, enabling its production from glucose directly. In this pathway, 4-hydroxyphenylpyruvate was converted to salvianic acid A via D-lactate dehydrogenase (encoding by d-ldh from Lactobacillus pentosus) and hydroxylase complex (encoding by hpaBC from E. coli). Furthermore, we optimized the pathway by a modular engineering approach and deleting genes involved in the regulatory and competing pathways. The metabolically engineered E. coli strain achieved high productivity of salvianic acid A (7.1g/L) with a yield of 0.47mol/mol glucose.

[Effects of tongfu decoction on rats' rapid gastric emptying].

OBJECTIVE: To observe the effects of Tongfu Decoction (TFD) on the gastric emptying of normal rats, thus exploring whether it could promote gastric emptying rapidly. METHODS: Thirty Sprague-Dawley (SD) rats were randomly divided into 3 groups, i.e., the normal control group, the domperidone group, and the TFD group, 10 in each group. They were respectively administered with normal saline, the domperidone suspension, and TFD by gastrogavage. Thirty min later the gastric emptying of mice was detected by single photon emission computed tomography technology (SPECT) labeled with 99m Tc-DTPA, and the gastric half-emptying time and the gastric emptying rate were calculated. RESULTS: The gastric half-emptying time was (19.0 +/-1.7) min in the normal control group, (12.9 +/- 3.4) min in the domperidone group, and (12.7 +/- 4.1) min in the TFD group. Compared with the normal control group, the gastric half-emptying time was significantly shortened in the domperidone group and the TFD group (P <0.05). The gastric emptying rate at 15 min was 41.1% +/- 5. 8% in the normal control group, 52.9% +/- 10.9% in the domperidone group, and 56.0% +/- 10.3% in the TFD group, while at 30 min it was 65.6% +/- 2.8%, 72.9% +/- 2.6%, and 72.4% +/- 4.9%, respectively. Compared with the normal control group, the gastric emptying rate at 15 min and 30 min both significantly increased in the domperidone group and the TFD group (P <0.05). There was no statistical difference in the gastric half-emptying time or the gastric emptying rate between the two groups (P >0.05). CONCLUSION: TFD showed similar effects as domperidone in rapidly promoting gastric emptying, and could shorten the gastric half-emptying time in normal rats.

Sofnolime with different water content causes different effects in two sevoflurane inhalational induction techniques with respect to the output of compound-A.

OBJECTIVE: During sevoflurane anesthesia with Sofnolime for CO(2) absorption, the factors affecting the production of compound A (a chemical is nepherotoxic) are still not clear. This study is designed to investigate the effects of different fresh gas flow during induction, the vital capacity induction (VCI) vs. the tidal volume breath induction (TBI) on the compound-A production with a fresh Sofnolime or a dehydrated Sofnolime using a simulated lung model. METHOD: The experiments were randomly divided into four groups: group one, VCIf, vital capacity fresh gas inflow with fresh Sofnolime; group two, TBIf, tidal volume breath fresh gas inflow with fresh Sofnolime; group three, VCId, vital capacity fresh gas inflow with dehydrated Sofnolime, and group four, TBId, tidal volume breath fresh gas inflow with dehydrated Sofnolime. The inspired sevoflurane was maintained at 8%, the concentrations of compound-A were assayed using Gas-spectrum technique, and Sofnolime temperatures were monitored at 1-min intervals throughout the experiment. RESULTS: The mean and maximum concentrations of compound A were significantly higher in the vital capacity group than the tidal volume breath group (P<0.01). At the beginning of anesthesia maintenance, the compound-A concentration in group VCIf was 36.28±6.13 ppm, which was significantly higher than the 27.32±4.21 ppm observed in group TBIf (P<0.01). However, these values decreased to approximately 2 ppm in the dehydrated Sofnolime groups. Sofnolime temperatures increased rapidly in the dehydrated Sofnolime groups but slowly in the fresh Sofnolime groups. CONCLUSION: With fresh Sofnolime, vital capacity induction increased compound-A production in the circuit system compared with tidal volume breath induction. However, with dehydrated Sofnolime, the effects of the two inhalation induction techniques on compound-A output were not significantly different.

Discovery and verification of gelsolin as a potential biomarker of colorectal adenocarcinoma in the Chinese population: Examining differential protein expression using an iTRAQ labelling-based proteomics approach.

OBJECTIVE: To identify and validate potential biomarkers of colorectal adenocarcinoma using a proteomic approach. METHODS: Multidimensional liquid chromatography/mass spectrometry was used to analyze biological samples labelled with isobaric mass tags for relative and absolute quantitation to identify differentially expressed proteins in human colorectal adenocarcinoma and paired normal mucosa for the discovery of cancerous biomarkers. Cancerous and noncancerous samples were compared using online and offline separation. Protein identification was performed using mass spectrometry. The downregulation of gelsolin protein in colorectal adenocarcinoma samples was confirmed by Western blot analysis and validated using immunohistochemistry. RESULTS: A total of 802 nonredundant proteins were identified in colorectal adenocarcinoma samples, 82 of which fell outside the expression range of 0.8 to 1.2, and were considered to be potential cancer-specific proteins. Immunohistochemistry revealed a complete absence of gelsolin expression in 86.89% of samples and a reduction of expression in 13.11% of samples, yielding a sensitivity of 86.89% and a specificity of 100% for distinguishing colorectal adenocarcinoma from normal tissue. CONCLUSIONS: These findings suggest that decreased expression of gelsolin is a potential biomarker of colorectal adenocarcinoma.

Management of procedural pain in the intensive care unit.

Pain is a common experience for inpatients, and intensive care unit (ICU) patients undergo more pain than other departmental patients, with an incidence of 50% at rest and up to 80% during common care procedures. At present, the management of persistent pain in ICU patients has attracted considerable attention, and there are many related clinical studies and guidelines. However, the management of transient pain caused by certain ICU procedures has not received sufficient attention. We reviewed the different management strategies for procedural pain in the ICU and reached a conclusion. Pain management is a process of continuous quality improvement that requires multidisciplinary team cooperation, pain-related training of all relevant personnel, effective relief of all kinds of pain, and improvement of patients' quality of life. In clinical work, which involves complex and diverse patients, we should pay attention to the following points for procedural pain: (1) Consider not only the patient's persistent pain but also his or her procedural pain; (2) Conduct multimodal pain management; (3) Provide combined sedation on the basis of pain management; and (4) Perform individualized pain management. Until now, the pain management of procedural pain in the ICU has not attracted extensive attention. Therefore, we expect additional studies to solve the existing problems of procedural pain management in the ICU.

Expert consensus on the monitoring and treatment of sepsis-induced immunosuppression.

Emerged evidence has indicated that immunosuppression is involved in the occurrence and development of sepsis. To provide clinical practice recommendations on the immune function in sepsis, an expert consensus focusing on the monitoring and treatment of sepsis-induced immunosuppression was developed. Literature related to the immune monitoring and treatment of sepsis were retrieved from PubMed, Web of Science, and Chinese National Knowledge Infrastructure to design items and expert opinions were collected through an online questionnaire. Then, the Delphi method was used to form consensus opinions, and RAND appropriateness method was developed to provide consistency evaluation and recommendation levels for consensus opinions. This consensus achieved satisfactory results through two rounds of questionnaire survey, with 2 statements rated as perfect consistency, 13 as very good consistency, and 9 as good consistency. After summarizing the results, a total of 14 strong recommended opinions, 8 weak recommended opinions and 2 non-recommended opinions were produced. Finally, a face-to-face discussion of the consensus opinions was performed through an online meeting, and all judges unanimously agreed on the content of this consensus. In summary, this expert consensus provides a preliminary guidance for the monitoring and treatment of immunosuppression in patients with sepsis.

Exploration of transmission chain and prevention of the recurrence of coronavirus disease 2019 in Heilongjiang Province due to in-hospital transmission.

The coronavirus disease 2019 (COVID-19) epidemic is a major public health emergency characterized by fast spread, a wide range of infections, and enormous control difficulty. Since the end of December 2019, Wuhan has become the first core infection area of China's COVID-19 outbreak. Since March 2020, the domestic worst-hit areas have moved to the Heilongjiang Province due to the increased number of imported COVID-19 cases. Herein, we reported the major COVID-19 outbreak, which caused a rebound of the epidemic in Harbin, China. After the rebound, different levels of causes for the recurrence of COVID-19, including city-level, hospital-level, and medical staff-level cause, were investigated. Meanwhile, corresponding countermeasures to prevent the recurrence of the epidemic were also carried out on the city level, hospital level, and medical staff level, which eventually showed the effect of infection control function in a pandemic. In this study, we described the complete transmission chain, analyzed the causes of the outbreak, and proposed corresponding countermeasures from our practical clinical experience, which can be used as a valuable reference for COVID-19 control.

Standardization of critical care management of non-critically ill patients with COVID-19.

The large global outbreak of coronavirus disease 2019 (COVID-19) has seriously endangered the health care system in China and globally. The sudden surge of patients with severe acute respiratory syndrome coronavirus 2 infection has revealed the shortage of critical care medicine resources and intensivists. Currently, the management of non-critically ill patients with COVID-19 is performed mostly by non-intensive care unit (ICU) physicians, who lack the required professional knowledge, training, and practice in critical care medicine, especially in terms of continuous monitoring of the respiratory function, intervention, and feedback on treatment effects. This clinical problem needs an urgent solution. Therefore, here, we propose a series of clinical strategies for non-ICU physicians aimed at the standardization of the management of non-critically ill patients with COVID-19 from the perspective of critical care medicine. Isolation management is performed to facilitate the implementation of hierarchical monitoring and intervention to ensure the reasonable distribution of scarce critical care medical resources and intensivists, highlight the key patients, timely detection of disease progression, and early and appropriate intervention and organ function support, and thus improve the prognosis. Different management objectives are also set based on the high-risk factors and the severity of patients with COVID-19. The approaches suggested herein will facilitate the timely detection of disease progression, and thus ensure the provision of early and appropriate intervention and organ function support, which will eventually improve the prognosis.

COVID-19 patient with an incubation period of 27 d: A case report.

BACKGROUND: As a highly contagious disease, coronavirus disease 2019 (COVID-19) is wreaking havoc around the world due to continuous spread among close contacts mainly via droplets, aerosols, contaminated hands or surfaces. Therefore, centralized isolation of close contacts and suspected patients is an important measure to prevent the transmission of COVID-19. At present, the quarantine duration in most countries is 14 d due to the fact that the incubation period of severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) is usually identified as 1-14 d with median estimate of 4-7.5 d. Since COVID-19 patients in the incubation period are also contagious, cases with an incubation period of more than 14 d need to be evaluated. CASE SUMMARY: A 70-year-old male patient was admitted to the Department of Respiratory Medicine of The First Affiliated Hospital of Harbin Medical University on April 5 due to a cough with sputum and shortness of breath. On April 10, the patient was transferred to the Fever Clinic for further treatment due to close contact to one confirmed COVID-19 patient in the same room. During the period from April 10 to May 6, nucleic acid and antibodies to SARS-CoV-2 were tested 7 and 4 times, respectively, all of which were negative. On May 7, the patient developed fever with a maximum temperature of 39℃, and his respiratory difficulties had deteriorated. The results of nucleic acid and antibody detection of SARS-CoV-2 were positive. On May 8, the nucleic acid and antibody detection of SARS-CoV-2 by Heilongjiang Provincial Center for Disease Control were also positive, and the patient was diagnosed with COVID-19 and reported to the Chinese Center for Disease Control and Prevention. CONCLUSION: This case highlights the importance of the SARS-CoV-2 incubation period. Further epidemiological investigations and clinical observations are urgently needed to identify the optimal incubation period of SARS-CoV-2 and formulate rational and evidence-based quarantine policies for COVID-19 accordingly.

Development and evaluation of diffusive gradients in thin films based on nano-sized zinc oxide particles for the in situ sampling of tetracyclines in pig breeding wastewater.

The pollution of antibiotics, including tetracyclines (TCs), in aquatic environments has become an issue of concern in recent years. Herein, an in situ sampling of TCs in pig breeding wastewater that utilizes the technique of diffusive gradients in thin films (DGT), based on commercial nanosized ZnO (nanoZnO) particles as the potential effective binding agent and a polyethersulfone (PES) membrane as the diffusion layer, was developed. The diffusion coefficients of tetracycline (TC), oxytetracycline (OTC) and chlortetracycline (CTC) in a PES membrane at 25 °C were (1.37 ±â€¯0.06) × 10-6 cm2 s-1, (1.29 ±â€¯0.05) × 10-6 cm2 s-1 and (1.94 ±â€¯0.07) × 10-6 cm2 s-1, respectively. The results showed that the adsorption capacities of a gel disc containing 2.5 g L-1 of nanoZnO particles were as high as 3.93 ±â€¯0.20 mg disc-1 for TC, 3.21 ±â€¯0.20 mg disc-1 for OTC and 4.62 ±â€¯0.22 mg disc-1 for CTC. Both a solution pH in the range of 5-9 and an ionic strength (as pNaCl) in the range of 1-3 had an insignificant influence on the TCs uptake by nanoZnO-DGT samplers. There was no significant influence of fulvic acid or tannic acid on the TC uptake by nanoZnO-DGT samplers at the tested mass ratios. For all spiked freshwater samples, there was no notable interference of matrices on the performance of the nanoZnO-DGT samplers, suggesting that the nanoZnO-DGT samplers yielded satisfactory results for the uptake of TCs at concentrations existing in the spiked freshwater samples. Field deployment of the nanoZnO-DGT samplers in pig breeding wastewater also exhibited excellent precision and accuracy, indicating that the nanoZnO-DGT samplers could be used as a promising method for the in situ sampling of TC antibiotics in aquatic environments.

Diffuse large B-cell lymphoma arising from follicular lymphoma with warthin's tumor of the parotid gland - immunophenotypic and genetic features: A case report.

BACKGROUND: Warthin's tumor (WT) is composed of several cysts that are lined with tall, bilayered oncocytic columnar cells and lymphoid stroma. Within WT, the two components rarely transform into carcinoma or lymphoma, and when it does, carcinoma is the most common type. Approximately 28 cases of lymphoma with WT have been reported, most of which were non-Hodgkin lymphomas, and only a few cases were Hodgkin lymphomas. In the present report, we studied a case of diffuse large B cell lymphoma (DLBCL) arising from follicular lymphoma (FL) with WT in the parotid gland and its immunophenotypic and genetic features. CASE SUMMARY: A 67-year-old man presented with a slowly enlarging right cheek mass for 12 years, and the mass began to change in size over a 2-mo time period. Over time, the patient felt mild local pain and right cheek discomfort. His medical history included a hepatitis B virus infection for 20 years and 30 years of smoking. Gross examination of the excised specimen showed a gray-red and gray-white appearance and a soft texture lobulated external surface neoplasm that measured 9 cm × 8 cm × 7 cm and was well circumscribed by relative normal parotid gland tissue. In cross section, the cut surfaces of the neoplasm were multicystic and had a homogeneous scaly appearance. A small fluid was discovered in the cyst. Bilateral oxyphilic, cuboidal or polygonal epithelium cells and lymphoid intraparenchymal components were observed. Many medium- to large-sized lymphoid cells were observed diffusely in part of the neoplasm, and a few secondary lymphoid follicles were observed at the center or edge of the neoplasm. Immunohistochemical staining showed that the columnar oncocytic cells were positive for AE1/AE3; neoplastic cells located in coarctate follicular were positive for CD20, Pax-5, bcl-2 and bcl-6; and the adjacent diffusely medium- to large-sized lymphoid cells were positive for Pax-5, bcl-6, CD20, MUM-1, bcl-2 and CD79a. The bcl-6 (3q27) break-apart rearrangement was observed, and an Epstein Barr virus test was negative in the tumor cells. The patient survived 6 months after being diagnosed without any treatment. CONCLUSION: WT-associated lymphoma is a very rare neoplasm in the parotid gland. Most cases are B cell non-Hodgkin lymphomas and involve middle-age and older males. This case highlights the extremely rare association of DLBCL arising from FL with WT and the importance of deliberate evaluation of the WT intraparenchymal stroma. Molecular detection techniques have potential advantages in the diagnosis of lymphoma with WT.

Uncoupling Protein 2 Drives Myocardial Dysfunction in Murine Models of Septic Shock.

Cardiac dysfunction is a major component of sepsis-induced multiorgan failure in critical care units. Uncoupling protein 2 (UCP2) involves immune response, regulation of oxidative stress, and maintenance of mitochondrial membrane potential as well as energy production. However, whether and how UCP2 plays roles in the development of septic cardiac dysfunction are largely unknown. Here, intraperitoneal injection of LPS significantly activated UCP2 expression accompanied by a significant decrease of cardiac function and caused a significantly lower survival rate in mice. Of note, knockdown of UCP2 through a cardiotropic adenoassociated viral vector carrying a short hairpin RNA (shRNA) specifically targeting the UCP2 evoked resistance to LPS-triggered septic cardiac dysfunction and lethality in vivo. Moreover, UCP2 deficiency ameliorated the reduced levels of intracellular ATP in the LPS-challenged heart tissues and preserved mitochondrial membrane potential loss in primary adult mouse cardiomyocytes in LPS-challenged animals. Mechanistically, we confirmed that the inhibition of UCP2 promoted autophagy in response to LPS, as shown by an increase in LC3II and a decrease in p62. At last, the autophagy inhibitor 3-MA abolished UCP2 knockdown-afforded cardioprotective effects. Those results indicate that UCP2 drives septic cardiac dysfunction and that the targeted induction of UCP2-mediated autophagy may have important therapeutic potential.

[17 beta-estradiol inhibits T-type calcium channels in mouse spermatogenic cells].

OBJECTIVE: To investigate the effect of 17 beta-estradiol (E2) on T-type calcium currents (ICaT) in spermatogenic cells. METHODS: Ca2+ currents were obtained in acutely dissociated mouse spermatogenic cells by the whole-cell patch clamp technique and the effects of E2 on ICaT were observed. RESULTS: E2 at the concentrations of 1, 10 and 100 micromol/L significantly inhibited ICaT in the mouse spermatogenic cells, with the KC50 value of 8.89 micromol/L and the inhibition rates of (13.48 +/- 1.86) %, (28.98 +/- 2.70) and (52.93 +/- 3.42)%, respectively (n = 6, P < 0.05). E2 of 100 micromol/L significantly changed the activation and inactivation of ICaT: the half activation potential (Va 1/2) and the activation steepness factor (Ka) from ( -48.94 +/- 0.22) mV and (5.19 +/- 0.19) mV to (-54.34 +/- 1.02) mV and (6.02 +/- 0.84) mV ( n = 5, P < 0.05) , and the half inactivation potential (Vi 1/2) and the inactivation steepness factor (Ki) from (-56.51 +/- 0.13) mV and (3.36 +/- 0.11) mV to (-61.78 +/- 0.50) mV and (4.25 +/- 0.37) mV, respectively (n = 5, P < 0.05). CONCLUSION: E2 has a significant inhibitory effect on ICaT in mouse spermatogenic cells in a con-centration-dependent manner.

[Experimental study on relationship between TCM syndrome type and gastric mucosa cell proliferation in rat model of chronic atrophic gastritis].

OBJECTIVE: To explore the relationship between TCM syndrome type and gastric mucosa cell proliferation related controlling gene protein in chronic atrophic gastritis (CAG). METHODS: Expressions of cell proliferation related controlling gene protein, including proliferative cell nuclear antigen (PCNA), epidermal growth factor receptor (EGFR) and c-myc, from gastric mucosa of CAG model rats with different syndrome types were measured by immunohistochemistry and the changes of them before and after TCM intervention were also analyzed by image analysis. RESULTS: Protein expressions of PCNA, EGFR and c-myc in gastric mucosa of CAG model rats with different syndrome types (Pi-deficiency type, Gan-stagnation type and dampness-heat type) were different to some extent, and all of them reduced significantly after TCM intervention in the model rats of all syndrome types. CONCLUSION: Expressions of cell proliferation related controlling gene protein in gastric mucosa of CAG model rats of different syndrome types were different to some extent, which provides a certain experimental evidence for revealing the essence of TCM syndrome type of CAG and judging the prognosis of various types.

[Effects of flunarizine on T-type calcium channels in mouse spermatogenic cells].

OBJECTIVE: To investigate the effects of Flunarizine (Flu) on T-type calcium currents (ICaT) in spermatogenic cells. METHODS: Ca2+ currents were obtained in acutely dissociated mouse spermatogenic cells using whole-cell patch clamp technique and the effects of Flu on ICaT were observed. RESULTS: Flu of 0.1, 1, 10, 100 micromol/L inhibited ICaT in mouse spermatogenic cells significantly with the K50 value of 0.289 micromol/L (P < 0.05). With the holding potential at -90 mV and stimulating potential at -30 mV, the inhibition rates of Flu were (23.34 +/- 2.76)%, (46.04 +/- 3.52)%, (62.52 +/- 3.70)% and (73.52 +/- 3.12)%, respectively. CONCLUSION: Flu has significant inhibitory effects on ICaT in mouse spermatogenic cells, with concentration dependence. Ca(v)3.2 is the main contributor to T-type Ca2+ currents in spermatogenic cells.

Diagnostic value of Wilms tumor 1 and CD44 in Langerhans cell sarcoma: case series of 4 patients.

Langerhans cell sarcoma (LCS) is a rare tumor with markedly malignant cytological features originating from Langerhans cells. LCS diagnosis is difficult and requires differentiation from other malignant tumors and Langerhans cell histiocytosis (LCH). Immunochemical antibodies, such as langerin, S-100 protein, and CD1a, have been used to diagnose LCS, but the results are crossed with LCH. To determine more significant biomarkers of LCS, we studied the expression and distribution pattern of Wilms tumor 1 (WT1) and cluster of differentiation 44 (CD44) in LCS. A broad panel of antibodies was used for immunohistochemical technology. Simultaneously, dual immunofluorescence staining examination and fluorescence in situ hybridization staining methods were used to study the location of WT1 and CD44 in LCS tumor cells. The results showed that tumor cells expressed WT1, CD44, and other special Langerhans cell markers (langerin, CD1a, and S-100 protein). LCS cells in all the cases showed normal cytogenetic findings without overexpression of WT1 and CD44. The expression of WT1 and CD44 was observed on langerin tumor cells by dual immunofluorescence staining examination in LCS. Our results suggest that WT1 and CD44 are potential biomarkers for LCS diagnosis. Clear understanding of their functional roles may further explain the pathogenesis of this highly malignant tumor and develop some novel immunotherapy strategies.