Induced pluripotent stem cells (iPSCs) can be propagated indefinitely, while maintaining the capacity to differentiate into all cell types in the body except for the extra-embryonic tissues. This iPSC technology not only represents a new way to use individual-specific stem cells for regenerative medicine but also constitutes a novel method to obtain large numbers of disease-specific cells for biomedical research. However, the low efficiency of reprogramming and genomic integration of oncogenes and viral vectors limit the potential application of iPSCs. Chemical-induced reprogramming offers a novel approach to generating iPSCs. In this study, a new combination of small-molecule compounds (SMs) (sodium butyrate, A-83-01, CHIR99021, Y-27632) under conditions of transient folate deprivation was used to generate iPSC. It was found that transient folate deprivation combined with SMs was sufficient to permit reprogramming from mouse embryonic fibroblasts (MEFs) in the presence of transcription factors, Oct4 and Klf4, within 25 days, replacing Sox2 and c-Myc, and accelerated the generation of mouse iPSCs. The resulting cell lines resembled mouse embryonic stem (ES) cells with respect to proliferation rate, morphology, pluripotency-associated markers and gene expressions. Deprivation of folic acid, combined with treating MEFs with SMs, can improve the inducing efficiency of iPSCs and reduce their carcinogenicity and the use of exogenous reprogramming factors.

ObjectiveTo evaluate the diagnostic value of “taller-than-wide sign” of thyroid malignancy in transverse and longitudinal plane using ultrasound.Methods708 thyroid lesions including 302 benign nodules and 406 malignant nodules ( 390 papillary carcinomas,12 medullary carcinomas,4 follicular carcinomas) that were diagnosed at surgery were included in this study.The sensitivity and specificity of “taller-than-wide sign” in transverse (A/TC ≥ 1 ) and longitudinal plane (A/TL ≥ 1 ) were evaluated using ultrasound.All thyroid malignant lesions were divided into three groups according to size of the nodules:group A≤1 cm; group B≤2 cm,group C＞2 cm.The sensitivity of each group was also evaluated.ResultsThe constituent ratio of “taller-than-wide sign” between benign and malignant nodules was significantly different (P＜0.001 ).The differences of sensitivity of A/TL ≥ 1 and A/TC ≥ 1 among three groups were also found( P ＜0.001),and the sensitivity of A/Tc≥1 was higher than A/TL≥1.ConclusionsA/TC≥1 found by ultrasound contributes to differentiate the thyroid carcinoma from benign nodules.For a large thyroid lesion,other indicators are needed to diagnose thyroid carcinoma.

The pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a serious impact on global public health and the economy. SARS-CoV-2 infiltrates host cells via its surface spike protein, which binds to angiotensin-converting enzyme 2 on the host cell membrane. As a result, small molecules targeting spike protein have emerged as a hotspot in anti-SARS-CoV-2 drug research. Activity screening is an important step in seeking small molecule drugs. Therefore, this article aims to review the biological activity evaluation methods of small molecule inhibitors targeting SARS-CoV-2 spike protein, with the goal of laying the foundation for the discovery of new anti-SARS-CoV-2 drugs.

To explore a new method for identification of Mongolian patent medicine (MPM) by PCR amplification of specific alleles. Eight kinds of MPM were used to study the identification of "Digeda" raw materials. The total DNA of Lomatogonium rotatum and Corydalis bungeana samples were extracted through modified CTAB method, psbA-trnH sequence was amplified by PCR and sequenced directionally. Specific primer was designed. The DNA of 8 kinds of MPM also was extracted and purified by the commercial DNA purification kits. The rbcL and two pair of specific primers sequences were amplified. The specific amplified products were sequenced in forward directions. All specific sequences were aligned and were analyzed. The results indicated that L rotatum can be identified by specific primers from Digeda-4 Tang, Digeda-8 San, Digeda-4 San, and C. bungeana medicinal materials can be identified by specific primers from Li Dan Ba Wei San, Yi He Ha Ri-12 and A Ga Ri-35. PCR amplification of specific alleles can stably and accurately distinguish raw medicinal materials in MPM.
Alleles ; DNA Primers ; genetics ; DNA, Plant ; genetics ; Medicine, Mongolian Traditional ; Molecular Sequence Data ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction ; methods

Objective To discuss the clinical phases of alkali chemical burn and the criteria of grading for severity of late phase.Methods Fourty-two patients with alkali burn of stable late stage were divided into four grades: mild (Grade Ⅰ), moderate (Grade Ⅱ), severe (Grade Ⅲ) and most severe (Grade Ⅳ). The selection of various operations depended on the grades. In mild patients, vascular membranous tissue and fibrous conjunctiva were resected. Autografts with stem cells and conjunctiva were transplanted simultaneously. Lamella was transplanted routinely using the fresh donor within 24 hours, with stem cells of limbus and corneal epithelium, combined with autograft stem cells of limbus implantation for Grade Ⅱ and Ⅲ. In most severe patients, three procedures were followed: monitoring and controlling intraocular pressure, reconstruction of limbus epithelium, and penetrating keratoplasty (PKP).Results All operations of 4 eyes of Grade Ⅰ were successful. Among 13 eyes of moderate grade, the transplantations of stem cells with antografts, corneal epithelium and lamellae with allografts (66.7% of PKPs) were successful. The visual acuity of 12 eyes was over 0.05. Among 10 eyes of severe grade, the transplantation of stem cells with autografts of 8 eyes was successful (80.0%). PKP of 6 eyes was successful (75.0%). The visual acuity of 9 eyes was over 0.05. Among 15 eyes of most severe grade, 6 were unilateral. The operation of 4 eyes was similar to that of the severe grade and it was successful. For 9 eyes of bilateral burn, stem cells transplantation and PKP with allografts were performed, but failed at 10 months postoperatively. The complications were treated. The visual acuity of 9 eyes was over 0.05.Conclusions The efficacy of late treatment of ocular alkali burn is dependent on several factors: unilateral or bilateral burn, the extent of damaged stem cells and corneal endothelium, the severity of complication. 70.6% of operations of unilateral burn, which was more severe than the moderate, were successful. But the treating result of severe bilateral burn was not satisfactory.

Objective: To evaluate the feasibility and efficacy of naked-eye assessment (NA) of ultrasound-guided fine needle aspiration cytology(US-FNAC) smears, which was performed by a trained non-cytological physician. Methods: A total of 290 smears of FNAC in 143 thyroid nodules were used to evaluate the value of NA by an assistant with more than two years experience of intervention with ultrasound guidance. NA results such as the background of smear (bloody/non-bloody), thickness (thick/thin), as well as the contents (granulated/non-granulated) were recorded. The correlation between NA and cytological results was analyzed. Number of cells under microscopy, the non-diagnostic rate, and the significance between benignity and malignancy with different features of specimens were compared. Results: There was no significant difference between the NA background and cytological findings(P=0.707, 0.200, 0.705, respectively). There was significance in number of cells, non-dianostic rate between thick and thinner specimens (P=0.000, 0.001, respectively). However, there was no significance in benignity and malignancy between thick and thinner specimens (P=0.385). There was significance in cytology results between the granulated and non-granulated smears(all P=0.000). The difference was found in 51.1% of the smears with granulates were malignancy in cytology. While 77.8% of the non-granulated smears were ultimately diagnosed as benignity. Conclusions The NA could be performed by a trained non-cytological physician and contribute to preliminary predictions of cytology effect.

Line scanning quantitative analysis method on silicate with small laser beam ( < 15 μm) was developed using laser ablation sector field inductively coupled plasma mass spectrometry (LA-SF-ICP-MS). Differences on signal intensity and elemental fractionation induced by different laser sampling patterns were compared. While spot ablation with small laser beam, the elemental signal intensity decreased with time significantly, and the elemental fractionation was obvious. In contrast, the elemental signal intensity by line scanning was higher and more stable and line scanning was free of elemental fractionation. Therefore, identical ablation pattern and condition should be used for the standard and the unknown sample in LA-ICP-MS quantitative analysis. A single pulse experiment was carried out to investigate the washout time when coupled to two-volume ablation cell. The result indicated that the elemental intensity decayed to the background value needed 2-3 s. The optimal parameters on SF-ICP-MS were set to reduce the effect of signal overlapping. Homogeneous sample KL2-G and titanite grains with composition zoning were analyzed by this method. Accurate element contents and element ratios indicated that fast washout time and optimal instrument parameters made it feasible to perform line scanning quantitative analysis accurately. Comparing to traditional microanalysis, line scanning quantitative analysis could reduce the laser beam size (<15 μm) and improve the spatial resolution efficiently. The potential of the technique to unveil compositional complexities in greater detail would help to improve our understanding of geochemical processes in mineral scale.

BACKGROUNDLaparoscopic Heller cardiomyotomy and Dor fundoplication is the surgical procedure of choice for esophageal achalasia. The aim of this study was to investigate the clinical outcome of laparoscopic Heller-Dor procedure in our initial series of 25 patients with achalasia.

METHODSBetween October 2003 and January 2006, a total of 25 patients with achalasia underwent laparoscopic Heller-Dor operation. Among them, 9 were male and 16 were female with an average age of (41.5 +/- 5.1) years (21-66). All the patients received upper gastrointestinal series (barium swallow), esophagogastroscopy, esophageal manometry to exclude esophageal carcinoma and to confirm the diagnosis, and 21 patients also had 24-hour ambulatory pH studies. All the patients were operated by laparoscopic modified Heller's myotomy with Dor fundoplication. In addition, 2 of them had combined laparoscopic cholecystectomy + excision of hepatic hemangioma and laparoscopic cholecystectomy, respectively.

RESULTSThe average operating time was (110.6 +/- 12.9) minutes (range, 60-180), operative blood loss averaged (18.6 +/- 7.1) ml (5-50), the median time to oral feeding was (1.6 +/- 0.4) days (1-4) and the median hospital stay was (12.6 +/- 1.2) days (10-20). There was no conversion to open surgery. Intraoperative mucosal perforation was encountered in six patients and was repaired in all of them by laparoscopic suture. All the patients had an uneventful recovery without postoperative complication. After a median follow-up of (10.6 +/- 7.2) months (1-27), 24 patients were asymptomatic and 1 had mild postoperative dysphagia.

CONCLUSIONSLaparoscopic Heller-Dor operation had the advantages of reduced compromise of the cardiopulmonary function, with less disruption of the supporting structures (phrenoesophageal membrane) of the antireflux mechanism, requiring simpler general anesthesia and providing excellent exposure permitting an easy fundoplication, less pain and reduced morbidity, shorter hospitalization and faster convalescence.

Adult ; Aged ; Digestive System Surgical Procedures ; methods ; Esophageal Achalasia ; surgery ; Esophagus ; surgery ; Female ; Follow-Up Studies ; Fundoplication ; methods ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods

OBJECTIVEBy means of phage-display technique, to screen polypeptides that specifically bind to human gastric cancer with high metastatic potential to peritoneum.

METHODSTwo human gastric cancer cell lines were used: GC9811-P with high metastatic potential to peritoneum and its wild type parental GC9811, to carry out subtractive screening with a phage display-12 peptide library.

RESULTSAfter three rounds of screening, 40 phage clones bond to GC9811-P cells were randomly selected. When injected into the peritoneal cavity of nude mice, 6 of the 40 clones did not bind to mouse peritoneum as examined by immunohistochemical staining. They were considered to be capable of binding specifically to GC9811-P cells. Sequence analysis revealed two different exogenous peptides: TLNINRLILPRT and SMSI(X)SPYI(XXX).

CONCLUSIONTwo peptides have been obtained that specifically bind to a gastric cancer cell variant GC9811-P, which easily disseminates to the peritoneum. Whether or not they could block GC9811-P metastasis to peritoneum in vivo remains to be determined.

Animals ; Binding Sites ; Cell Line, Tumor ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Peptide Library ; Peptides ; metabolism ; Peritoneal Neoplasms ; secondary ; Protein Array Analysis ; methods ; Protein Binding ; Sensitivity and Specificity ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVETo detect serve acute respiratory syndrome-associated coronavirus (SARS-CoV) and SARS-like-CoV in fruit bats captured in Guangzhou and its vicinity.

METHODSTotally 927 bats of 9 species (Cynopterus sphinx, Rousettus leschenaulti, Miniopterus schreibersi, Hipposideros pratti, Rhinolophusasinicus, Scotophilusakuhlii, Hipposideros Pomona, Rhinolophus affinis, and Rhinolophus pusillus) captured in Guangzhou and its vicinity from September 2004 to November 2005 were available for this investigation, from which 3,043 samples (813 throat swasb, 524 sera, 853 lung tissues and 853 colorectal tissue specimens) were obtained. SARS-Cov and SARS-like-CoV were detected in these specimens using diagnostic kit for novel coronavirus N protein (ELISA), SARS-CoV Virus RNA detection kit, fluorescence PCR, Genchip, RT-PCR and cell isolation culture methods.

RESULTS AND CONCLUSIONNo SARS-CoV and SARS-like-CoV were detected in the 3043 samples, indicating the current absence of SARS-CoV and SARS-like-CoV in the bats captured in Guangzhou and its vicinity.

Animals ; China ; epidemiology ; Chiroptera ; virology ; Disease Vectors ; Enzyme-Linked Immunosorbent Assay ; Humans ; Nucleocapsid Proteins ; metabolism ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; isolation & purification ; metabolism ; Severe Acute Respiratory Syndrome ; epidemiology ; transmission ; virology