Thirty-one patients with middle ear effusion were evaluated for the presence of chlamydia pneumonia by using polymerase chain reaction (PCR) assay. The presence of chlamydia neumonia was 19. 3% (6/31). The results indicated that PCR assay is a highly sensitive and specific for the detection of chlamydia pneumonia in the middle ear effusion and chlamydia pneumonia may be an aetiological agent of middle ear effusion.

Objective To investigate the mechanism of contrast-induced nephropathy (CIN) caused by Iopromide.Methods Two-four female SD rats were randomly divided into two groups which were control group and CIN group .The rats in CIN group were injected Io-promide via caudal vein ,the rats in control group were injected the equal amount of solvent .After 24 hours,all the rats were euthanized and tested.The excretion of 24 h urinary protein was detected using biochemistry assay .The expression of related cell cycle regulatory protein such as P21,P27 and TGF-β1 in glomerular visceral epithelium cells were measured using immunohistochemical technique .A semiquantitative score was used to evaluate the injure degree of glomerular and tubulointerstitium .Renal glomerular cell apoptosis was evaluated by TUNEL . Results Compared with control group ,CIN group rat glomerular epithelial cells of P 21,P27 and TGF beta 1 positive expression rate signifi-cantly increased,[(12.86 ±0.98) %vs (0.46 ±0.21)%,P=0.004 5],[(21.76 ±2.75)% vs (9.57 ±1.86)%,P =0.0071], [(12.85 ±5.54) vs (7.63 ±0.84),P=0.003 7)] respectively,24 h urine protein significantly increase [(23.44 ±5.22) mg/d vs (2.13 ±0.52) mg/d,P=0.007 0,P=0.005 0],CIN pathological damage of rat glomerular epithelial cells and apoptotic rate significantly more serious [(52.5 ±6.4)%vs (4.2 ±0.3) %,P =0.007 5].In addition,the renal pathologic scores were positively correlated with the excretion of 24hr urinary protein and the expression of P 21,P27,and TGF-β1(r=0.765,0.701,0.842,0.651,P<0.01).Conclusion Io-dine amine via increased glomerular epithelial cells P 27 and TGF-beta 1expression and urinary protein excretion , aggravating pathological damage and apoptosis .

Aim To investigate the proliferative effect and the apoptosis of rat epididymal epithelial cells induced by podophyllotoxin and its underlying mechanisms.Methods Primary epididymal epithelial cells were cultured in vitro.CCK-8 assay was used to analyze proliferation of epididymal epithelial cells induced by podophyllotoxin on 24, 48 and 72 h.The ultra structural changes of the epididymal epithelial cells were observed by transmission electron microscope.AnnexinV-FITC/PI staining was used to quantify the percentages of apoptosis in the total cell population.The TdTmediated dUTP nick end labeling(TUNEL) technique was applied to observe the morphological changes of apoptotic cells.The expression of tumor necrosis factor alpha(TNF-α) mRNA was investigated by real-time RT-PCR.The level of TNF-α in cell culture supernatant was measured by enzyme-linked immunosorbent assay(ELISA) technology.Western blot was per-formed to determine the protein expression of cytochrome C, caspase-3, caspase-8, caspase-9.Results Podophyllotoxin significantly inhibited the activity of proliferation and induced apoptosis of epididymal epithelial cells in a dose-and time-dependent manner(P<0.05), with a 50％ inhibitory concentration(IC50) value and corresponding 95％ confidence intervals(CI) of 59.36(15.50～227.41), 0.37(0.080～1.70), 0.077(0.017～0.35) μmol·L-1 at 24, 48 and 72 h, respectively.Podophyllotoxin induced cell volume turned round and cell nuclear fragmentation and mitochondrial vacuolation.RT-PCR and ELISA results showed that podophyllotoxin improved the expression of TNF-α mRNA and protein.Western blot results demonstrated that podophyllotoxin activated the protein expression of cytochrome C, caspase-3, caspase-8 and caspase-9.Conclusion Podophyllotoxin can induce rat epididymal epithelial cell apoptosis through both the mitochondria-regulated intrinsic pathway and the TNF receptor-mediated extrinsic pathway.

Objective To explore the neuroprotection of Shuxuetongmai capsule pretreatment, and the effect on the expression of p38 mitogen-activated protein kinase (p38MAPK) in rats with middle cerebral artery occlusion. Methods Ninety-six male SD rats were divided randomly into sham-operated group,ischemia/reperfusion group (I/R),ischemia preconditioning group (IP),and Shuxuetongmai group(n=24). Each group was further randomly divided into 4 subgroups by 3 h, 6 h, 24 h and 72 h after reperfusion, 6 rats in each subgroup. Sham-operated group was only performed artery separation . The middle cerebral artery occlusion (MCAO) model was set up in I/R rats by Longa method. The IP rats were performed for three minutes on the bilateral carotid artery ligation, and formed MCAO model 24 hours later. The rats in the Shuxuetongmai group were pretreated with Shuxuetongmai capsules for 14 days on gavage before the establishment of MCAO model. The neurological deficits were graded in rats by Zea Longa method. Western Blot was used to determine the protein expression of p38MAPK and P-p38MAPK. Tunel method was applied to detect the apoptosis of neurons and the relationship between expression of p38MAPK, P-p38MAPK and apoptosis of neuron. Results No neurological dysfunction appeared in the sham-operated group at each time points, but not for the other groups, which reached the peak at 24 h. Compared with the I/R group, IP group and Shuxuetongmai group presented the mild neurologic function deficiency at different time points in rats (P<0. 05), and no significant differences occurred between ischemia preconditioning group and Shuxuetongmai group (P>0.05). The obvious variation of the value of P-p38MAPK/p38MAPK wasn't detected in sham-operated group at different time points, while obviously presented in I/R group, and the ratios of P-p38MAPK/p38MAPK were increased gradually followed with reperfusion, approaching to the highest level at 24 h. Compared with the I/R group, the P-p38MAPK/p38MAPK declined from 3 h and to the lowest level at 24 h of reperfusion, in both IP and Shuxuetongmai groups(P<0. 05), and with similar phosphorylation. At different time points,very few neurons apoptosis were detected in sham-operated groups, but which increased gradually after reperfusion in other groups, and reached to the peak at 24 h. The neurons apoptosis in both IP group and Shuxuetongmai group were less than that in IR group ( P<0. 05 ) at different time points, and it showed no significant differences on neurons apoptosis between ischemia/preconditioning group and Shuxuetongmai group in rats (P>0. 05). Conclusion Shuxuetongmai capsule pretreatment can induce brain ischemic tolerance, attenuate the apoptosis of neurons in cerebral ischemia reperfusion, and improve neurologic function. The mechanism may be related to the inhibition of p38MAPK phosphorylation.

Objective To investigate the changes of coagulatory function in septic rats induced by cecal ligation and puncture(CLP). Methods Cecal ligation and puncture(CLP)were performed to induce sepsis in SD rats. Coagulation indexes were detected at 8,16 and 48 h after operation, and histopathological changes of the lung, kidney, liver and spleen were examined using HE staining. Results The 12-day survival rate of the CLP-induced septic rats was 30%,with an acute onset and high mortality. In the acute phase of disease development of the CLP rats, the activated partial thromboplastin time(APTT)was prolonged(P<0.05)at 8 h,the prothrombin time(PT)was prolonged at 16 h (P<0.05), the factor XII activity in the endogenous coagulation pathway and the factor VII activity in the extrinsic coagulation pathway showed a transient inhibition, the thrombin time(TT)was prolonged at 48 h(P<0.01), and the content of fibrinogen(FIB)was increased gradually from 16 h(P<0.001). Among the other important coagulation and anticoagulation indexes,the number of platelets(PLT)was decreased gradually from 8 h(P<0.01),while the number of vWF:Ag increased gradually from 8 h(P<0.001). The D-dimer amount gradually increased from 16 h(P<0.05),and the amount of PS:Ag significantly decreased until 48 h(P<0.001). However, there was no significant change in the antithrombin-III(AT-Ⅲ)content. The histopathological examination showed that there are different degrees of damages in the lung,kidney,liver and spleen tissues,but no obvious venous thrombosis and bleeding were found. Conclusions In the acute phase,there is coagulatory dysfunction in the septic rats,however,no histopathological changes such as venous thrombosis and bleeding were observed in the lung,kidney,liver and spleen tissues due to coagulatory dysfunction.

The Huangdi Neijing proposes the " using bitter herbs to nourish or purge" theory to guide clinical prescription and formulation of herbal remedies based on the physiological characteristics and functions of the five zang viscera, along with the properties and flavors of medicinal herbs. This study explored diabetic kidney disease pathogenesis and treatment based on the " using bitter herbs to nourish or purge" theory. Kidney dryness is a key pathological factor in diabetic kidney disease, and the disharmony of kidney dryness is an essential aspect of its pathogenesis. Strengthening is the primary therapeutic principle, and kidney dryness is a persistent factor throughout the occurrence and progression of diabetic kidney disease. In the early stage, the pathogenesis involves heat-consuming qi and injuring yin, leading to kidney dryness. In the middle stage, the pathogenesis manifests as qi deficiency and blood stasis in the collaterals, resulting in turbidity owing to kidney dryness. In the late stage, the pathogenesis involves yin and yang deficiency, with kidney dryness and disharmony. This study proposes the staging-based treatment based on the " need for firmness" characteristic of the kidney. The aim is to provide new insights for clinical diagnosis and treatment in traditional Chinese medicine by rationally using pungent, bitter, and salty medicinal herbs to nourish and moisturize the kidney. This approach seeks to promote precise syndrome differentiation and personalized treatment for different stages of diabetic kidney disease, thereby enhancing clinical efficacy.

【Objective】 To investigate the effects of one-stage additional posterior pedicle screws （PPS） internal fixation on early Cage subsidence after oblique lateral interbody fusion （OLIF）. 【Methods】 We made a retrospective analysis of 118 patients with lumbar degenerative diseases treated with OLIF at the Department of Orthopedics， Sir Run Run Shaw Hospital， School of Medicine， Zhejiang University， from January 2016 to December 2019. We divided the patients into OLIF stand-alone group （58 ones） and OLIF with PPS fixation group （60 ones） according to the surgical procedure. All the patients had preoperative frontal and lateral radiographs of the lumbar spine， and CT and MR scans were performed. The clinical outcomes and reoperation rates of the two groups were compared at immediate postoperative follow-up and at 1， 3， 6 and 12 months. X-ray and CT examinations were performed to assess Cage subsidence in both groups at each postoperative follow-up. 【Results】 There was no statistical difference between the two groups in baseline data and surgical segmentation. Of the 118 patients with 141 discs who underwent OLIF surgery， 58 patients with 68 discs received OLIF stand-alone surgery and 60 ones with 73 discs received OLIF with PPS fixation. There were no significant differences in intraoperative bleeding， complications， or postoperative clinical outcomes between the two groups （P>0.05）， and the Cage subsidence rate was 22.4% in OLIF stand-alone group and 5% in OLIF with PPS fixation group， with significant difference between the two groups （P<0.01）. 【Conclusion】 Both OLIF stand-alone and OLIF additional PPS fixation can achieve good early clinical outcomes， and first-stage additional PPS fixation can significantly reduce the occurrence of Cage subsidence in the early postoperative period after OLIF.