Objective To construct a bicistronic expression vector containing HPV type 6b L1 gene, to express the recombinant vector in mammalian cells, and to establish a cell strain stably expressing HPV6b L1 gene. Methods After endonuclease digestion and purification, the gene fragment of HPV6b L1 was cloned into the eukaryotic expression vector pIRES2-enhanced green fluorescent protein (EGFP). The identification of the recombinant was realized via endonuclease digestion and sequence analysis. Then, the recombinant plasmid pIRES2-HPV6bLl-EGFP was transfected into NIH3T3 (a mouse embryonic fibroblast cell line) cells. Subsequently, the expression of EGFP was observed by fluorescent inverted microscopy, and HPV6b L1 mRNA expression by reverse transcription (RT)-PCR. Results The recombinant plasmid pIRES2-HPV6bLl-EGFP was successfully constructed, transfected into N1H3T3 cells, and selected by G418. The expression of EGFP was seen under an inverted fluorescence microscoy. RT-PCR proved the expression of HPV6b LI mRNA in transfected cells. Conclusions The recombinant plasmid pIRES2-HPV6bLl-EGFP was successfully constructed and transfected into NIH3T3 cells. Inverted fluorescent microscopy and RT-PCR confirmed the successful expression of HPV6b L1 in NIH3T3 cells.

BACKGROUND AND OBJECTIVECybeKnife is a newly developed technology in the field of stereotactic radiosurgery/radiotherapy (SRS/SRT). Compared with conventional SRS/SRT, there are many advantages for CyberKnife in terms of treating tumors that move with respiration, being real-time image-guidance, frameless, high accurateness, and so on. Recently, it has been used to treat different types of malignant carcinoma including intracranial and caudomedial tumors. This study was designed to evaluate the short-term efficacy and toxicity of the CyberKnife radiotherapy for locally advanced pancreatic cancer.

METHODSA total of 20 patients with locally advanced (stage II-III) pancreatic cancer treated with CyberKnife were recruited between April 2009 and December 2009. Of 20 patients, 13 were with cancer located at the pancreatic head and 7 were located at the pancreatic body and tail. The planning target volume (PTV) was defined as gross tumor volume (GTV) plus 2-3 mm, and more than 95% PTV should be covered by 75% isodose surface. The median of PTV was 47 cm³ (26-64 cm³). The median total prescription dose was 40 Gy (32-55 Gy) at 3-6 fractions. During treatment delivery, X-Sight Spine Tracking System was used in 5 patients to track movement of the tumor. Other 15 patients were implanted fiducials in the tumors to track movement of the tumor and patient breathing patterns.

RESULTSThe median follow-up time was 7 months (3-11 months). All patients had finished the treatment and 19 were alive by the last follow-up. Slight fatigue was the most common complain. Evaluated by CT scan, 6 were complete response, 9 were partial response, 3 were stable disease, and 1 was progression; 1 was dead. There were 6 patients with grade I granulocytopenia, 7 with grade I nausea, and 5 with grade II vomiting.

CONCLUSIONSThe CyberKnife radiosurgery for the locally advanced pancreatic cancer shows a high rate of local control and minimal toxicity. Long-term follow-up is necessary to evaluate the survival and late toxicity.

Adult ; Aged ; CA-19-9 Antigen ; blood ; Female ; Follow-Up Studies ; Humans ; Leukopenia ; etiology ; Male ; Middle Aged ; Nausea ; etiology ; Neoplasm Staging ; Pancreatic Neoplasms ; diagnostic imaging ; surgery ; Quality of Life ; Radionuclide Imaging ; Radiosurgery ; adverse effects ; Radiotherapy Dosage ; Remission Induction ; Thrombocytopenia ; etiology

Objective To investigate clinical and epidemiological features of pneumocystis jirovecii pneumonia (PJP) in kidney transplant recipients. Methods Clinical data of 68 kidney transplant recipients admitted from July, 2021 to December, 2021 were collected. All patients were divided into the PJP group (n=11), common pulmonary infection group (n=24) and non-pneumonia group (n=33) according to the status of pulmonary infection. The incidence and treatment of PJP after kidney transplantation were analyzed. Basic characteristics and laboratory parameters of the recipients were compared among all groups. The genotyping and transmission map of PJP patients were evaluated. Results Among 64 kidney transplant recipients, 11 cases were definitely diagnosed with PJP. The most common clinical manifestations included elevated body temperature, and dry cough complicated with progressive dyspnea. Chest CT scan showed diffuse interstitial inflammation and ground glass-like lesions of bilateral lungs in all patients. After diagnosis, all patients were orally given with compound sulfamethoxazole for 3-4 weeks. Two patients received non-invasive ventilator-assisted ventilation due to severe lung infection and dyspnea, and the remaining patients were given with nasal cannula oxygenation. One patient experienced elevated serum creatinine level upon discharge, and developed renal allograft failure. The remaining 10 recipients with PJP obtained normal renal allograft function, and no recipient died. Compared with the non-pneumonia group, the rejection rate was higher, the length of hospital stay was longer, the lymphocyte count was less, the lymphocyte proportion was lower, the levels of C-reactive protein, serum creatinine and lactate dehydrogenase were higher, and the levels of serum albumin was lower and CD4⁺T cell count was less in the PJP group (all P < 0.05). Compared with common pulmonary infection group, the lymphocyte count was less, the lymphocyte proportion was lower, the CD4⁺T cell count was less and 1, 3-β-D- glucan (BDG) level was higher in the PJP group (all P < 0.05). No new genotype was detected in 10 of the 12 testing samples. It was considered that PJP mainly depended on two transmission chains and two independent transmission individuals. Conclusions Kidney transplant recipients are prone to pneumocystis jirovecii (PJ) infection due to impaired cellular immune function. The most common clinical manifestations consist of elevated body temperature and dry cough complicated with progressive dyspnea, accompanied by headache and fatigue in partial patients. Chest CT scan shows diffuse interstitial inflammation and ground glass-like lesion of bilateral lungs. PJ may be transmitted through respiratory tract. Small-scale PJP might occur in the follow-up outpatient of kidney transplant recipients. Preventive measures should be delivered in a timely manner.

Objective To study the effect of Danshiliuhao decoction on the expression of tumor necrosis factor -α( TNF -α) and nuclear factor-κB( NF -κB ) in the bile pigment with stone in Guinea pigs. Methods The female Guinea pigs were randomly divided into three groups.The blank group ( n =10 ) , the model group ( n =15 ) and test group ( Danshiliuhao,n=15 ) .The latter two groups of the Guinea pigs were both modeled as the bile pigment stones model by feeding them with a different diet .The Guinea pigs in test group were given Danshi-liuhao decoction 7.7 g? kg-1? d-1 for 30 days.The rate of stone formation and the expression of the gallbladder TNF-αand NF-κB were observed af-ter administration by immunohistochemistry.Results The rate of stone formation was 90.62% in model group, significantly higher than that (26.76%) in the blank group.The results suggested that the model for Danshiliuhao is successfully built and the drug is an effective agent for anti-gallstones.The expression of the TNF -αand NF -κB in test group were significantly lower than model group. The difference was statistically significant ( P<0.05 ) .Conclusion Danshiliuhao Decoc-tion significantly reduced the rate of pigment stone formation in Guinea pigs.This may be related to the decreased expression of TNF-αand NF-κB proteins in gallbladders of Guinea pigs by the drug.

Objective To analyze the molecular characteristics of the newly isolated two Japanese encephalitis virus strains(JEV) in Wuhan. Methods The mosquitoes were collected in Wuhan from April to October in 2009. The envelope (E) protein gene of JEV was detected using RT-PCR and sequenced.Sequence comparisons and phylogenetic analysis were conducted using DNAstar and MegAlign. Results Two Japanese encephalitis virus (JEV) strains (WHJX09-9, WHJX09-10 ) were isolated from Culex tritaeniorhynchus among 16 mosquito pools and identified as genotype I. The result showed that the homology of the two strains was 98. 9％ in nucleotides and 100％ in deduced amines. The comparison between the new genotype 1 JEV strains and live attenuated vaccine strain SA14-14-2 in E gene showed that the homology of nucleotide sequence was 87.4％ and 87.9％ ,the homology of amino acid was 96. 9％ (totol 15 amino acid were different) in E gene. The mutation sites of amino acid distributed among three different coding domain,but no antigen binding site and neurotoxin-involved site of amino acid were changed. Conclusion Wuhan had appeared a new genotype of JEV which was different from the former strain isolated in Wuhan,the new JEV strains still had neurotoxicity but had high homology with the vaccine strains adopted in Wuhan. The vaccine could still be adopted to prevent Japanese encephalitis if steps were take to eradicate mosquitos at the same time. laboratory surveillance were also an important task to build an early-warning mechanism against JEV.

AIM:To investigate the inhibitory effect of corticosterone(CORT)on lipopolysaccharide(LPS)-induced expression of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)and its relation with xan-thine oxidase(XO).METHODS:An inflammatory model of mouse macrophage RAW 264.7 was established by stimula-ting with LPS.Total cellular protein was extracted after the macrophages were treated with CORT at different concentrations (0~900 μg/L).The protein levels of NLRP3 and caspase-1 were determined by Western blot.According to the treat-ments,the macrophages were divided into control group,LPS group,LPS+CORT group and LPS+allopurinol group.Cell components were extracted at 0,0.5,1,1.5 and 2 h.The protein levels of NLRP3 and XO were determined by Western blot,and the mRNA expression of NLRP3 and XO was detected by real-time PCR.RESULTS: CORT at 700 μg/L and above significantly inhibited the expression of NLRP 3 and the activation of caspase-1 in the macrophages induced by LPS (P<0.05).Compared with LPS group, the expression of NLRP3 and XO in LPS +CORT group was inhibited(P <0.05),and the expression of NLRP3 in LPS+allopurinol group was also reduced(P<0.05).CONCLUSION: High concentration of CORT inhibits the expression of NLRP 3 in LPS-induced mouse macrophages,which is associated with XO. The inhibitory effect of CORT may be related to the reduction of XO expression.

Objective To study the damage on organs from salt sensitivity hypertension or non-salt-sensitive hypertension and the selection of drug combination.Methods 120 hypertensive patiems including 60 cases salt-sensitive (SS) and 60 non-salt-sensitive (NSS) groups were selected in our hospital and their salt load tested.These two groups were randomly divided into two groups,each group with 30 patients,one was given felodipine and perindopril and the others were given indapamide sustained release tablets and peridopril to facilitate the 12-week treatment.Before and after the treatment,patients were tested for physiological indicators,such as sitting blood pressure,24-hour ambulatory blood pressure,insulin resistance index,comparing changes of various sub-index etc.Results Significantly different were seen in indices as fasting blood glucose and serum creatinine (P＜ 0.01),fasting insulin,left ventricular mass index,urinary albumin,body mass index,insulin resistance indices,while between the SS group and the NSS group(P＜0.05).In the SS group,when patients with various sub-indicators were using perindopril combined with indapamide treatment,the related detected indicators tended to be normal and with statistically significant differences (P＜0.05).In the NSS group,those related indexes also tended to be more normal when using felodipine combined with perindopril.However,there were statistically significant differences between the two groups (P＜0.05).Conclusion On SS hypertensive patients with target organ damages,perindopril and indapamide seemed to be more effective in NSS patients,indicating that the use of perindopril and felodipine combination,seemed to be more suitable.

OBJECTIVEStandardization of and training in adult male circumcision can significantly reduce its complication rate. Currently no such program exists for its standardization and training, making it difficult to guarantee the quality of male circumcision services. We therefore established a standardized surgical protocol for adult male circumcision in China using the Shang Ring, and applied it to a clinical study examining the performance of the Shang Ring in adult male circumcision.

METHODSA total of 328 adult men aged 18-58 (mean 27.8) years, 25 with phimosis and 303 with redundant prepuce, underwent circumcision with the Shang Ring, and evaluation of the operation time, pain scores (using the visual analog scale), postoperative complications, time for wound healing, and their satisfaction with the postoperative appearance.

RESULTSThe operation time was 4.7 +/- 1.3 minutes. The pain scores were 0. 2 +/- 0.6 during the surgery, 1.6 +/- 1.0 twenty hours postoperatively, 1.7 +/- 1.1 twenty hours prior to the ring removal, and 2.7 +/- 1.4 during the ring removal. Complications included infection in 2 (0.6%), bleeding in 2 (0.6%), and wound dehiscence in 2 (0.6%) of the patients. None of the patients with wound dehiscence required postoperative suturing and all were managed conservatively instead. Sixteen of the patients (4.9%) experienced penile edema. The time for complete wound healing after circumcision was 20.3 +/- 6.7 days. The rate of the patients'satisfaction was 99.7% (327/328).

CONCLUSIONThe standard protocol of adult male circumcision with the Shang Ring has the advantages of short operation time, slight pain, low rate of complications, and high satisfaction and acceptance of the patients. Strict standardization of the surgical protocol can maximize its clinical advantages for adult male circumcision.

Adolescent ; Adult ; China ; Circumcision, Male ; methods ; standards ; Humans ; Male ; Middle Aged ; Phimosis ; surgery ; Reference Standards ; Young Adult

BACKGROUNDPrior research about N400 has been mainly based on English stimuli, while the cognitive processing of Chinese characters is still unclear. The aim of the present study was to further investigate the semantic processing of Chinese idioms.

METHODSEvent related potentials (ERP) component N400 was elicited by 38 pairs of matching (congruent) and mismatching (incongruent) ended Chinese idioms: ending words with same phoneme but different shape and meaning (sPdSdM), with similar shape but different phoneme and meaning (sSdPdM), with same meaning but different phoneme and shape (sMdPdS), and words with different phoneme, shape and meaning (dPdSdM) and recorded by Guangzhou Runjie WJ-1 ERP instruments. In 62 right-handed healthy adults (age 19 - 50 years), N400 amplitudes and latencies were compared between matching and mismatching conditions at Fz, Cz and Pz.

RESULTSN400 showed a midline distribution and could be elicited in electrodes Fz, Cz and Pz. The mean values of N400 latencies and amplitudes were obtained for matching and mismatching ending words in healthy adults. Significant differences were found in N400 latencies and amplitudes in matching and mismatching ending-words idioms in healthy adults (P < 0.05). Compared with matching ending-words idioms, N400 latencies were prolonged and the amplitudes were increased in mismatching ones. N400s elicited by different types of stimuli showed different latencies and amplitudes, and longest N400 latency and largest N400 amplitude were elicited by ending-words with dPdSdM. No gender difference was found of N400 latency and amplitude in this study (P > 0.05).

CONCLUSIONSCompared with English stimuli, Chinese ideographic words could provide more flexible stimuli for N400 research in that the words have 3-dimension changes - phoneme, shape and meaning. Features of N400 elicited by matching and mismatching ending words in Chinese idioms are mainly determined by the meaning of the word. Some issues of N400 elicited by Chinese characters deserve further research.

Adult ; Cognition ; Evoked Potentials ; physiology ; Female ; Humans ; Male ; Middle Aged ; Reaction Time ; Reading ; Semantics ; Sex Characteristics

Clinical pharmacist participated in the whole process of combined drug treatment and medication monitoring of a patient with refractory edema secondary to end-stage diabetic kidney disease.By using dapagliflozin combined with sacubitril valsartan and conventional diuretics,the patient's body weight decreased by 16 kg,blood pressure returned to normal,systemic edema subsided and the symptoms of heart failure obviously improved.In this case,clinical pharmacist provided individualized pharmaceutical care and assisted clinicians to make a scientific and reasonable drug therapy,so as to provide a new idea and medication reference for clinical patients to delay the time of renal replacement therapy.