Preliminary study on the correlation between primary lacrimal punctal stenosis and epiphora by using AS-OCT / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To quantify the lacrimal punctum using anterior segment optical coherence tomography(AS-OCT)and analyze the correlation between the epiphora and primary punctal stenosis.METHODS: A cross-sectional study. There were 43 healthy volunteers(57 eyes)and 44 patients(62 eyes)with primary lacrimal punctal stenosis and epiphora enrolled in the Lacrimal Center of the Third Medical Center of PLA General Hospital from September 2020 to January 2021. The inferior punctum of all subjects were scanned by AS-OCT, and the images were quantified in order to observe the AS-OCT image features. The outer punctal diameter, punctal depth, tear well diameter and tear well depth were all measured simultaneously. The inferior punctum's punctal reserve and punctal reserve ratio were estimated according to the available parameter data.RESULTS: 1)The measured parameters conforming to normal distribution were expressed as mean±standard deviation. The observation group's outer punctal diameter, tear well diameter, tear well depth, and punctal reserve ratio were 458.19±63.58μm, 200.34±84.69μm, 188.95±87.50μm and 0.32±0.18, respectively, while the control group's outer punctal diameter, tear well diameter, tear well depth and punctal reserve ratio were 655.53±82.62μm, 230.26±107.02μm, 275.30±144.34μm, 0.46±0.23, respectively. The parameters in observation group were all lower than those of control group(P<0.05); 2)Parameter data that did not conform to normal distribution were expressed as medians and quartiles, and the results were as follows: the punctal depth in the observation group was 265.50μm(interquartile range 204.25-328.77)μm and the punctal reserve was 71.53μm(interquartile range 46.12-111.37)μm, respectively, while the punctal depth in the control group was 468.76μm(interquartile range 420.50-588.88)μm and the punctal reserve was 182.16μm(interquartile range 131.36-309.84)μm. This difference was statistically significant(P<0.05); 3)In the observation group, there was a negative correlation between the epiphora and the outer punctal diameter(r=-0.448, P<0.05), a positive correlation between the epiphora and tear well depth(r=0.335, P<0.05), and a negative correlation between the epiphora and the punctal reserve and punctal reserve ratio coefficient(r=-0.520, -0.566, P<0.05).CONCLUSION: The AS-OCT can aid in enhancing punctal morphology cognition; The outer punctal diameter and tear well depth are related to the epiphora; The primary punctal stenosis patients with smaller outer punctal diameters are more likely to have the symptom of epiphora, and in patients with punctal stenosis, the greater the potential capacity of the punctum to hold tears, the lesser the degree of epiphora.

Research model integrating "both medicinal and adjuvant properties" for essential oils from Chinese medicinal: based on characterization of medicinal properties / 中国中药杂志

Essential oils(EOs) from Chinese medicinals, which can be used as adjuvants and exert certain therapeutic effect, are directly used in Chinese medicine formulas. Conventional research strategy for EOs from Chinese medicinals is to compare the efficacy of the prescriptions before and after the addition of EOs, and the penetration-enhancing mechanisms of EOs remain unclear. In modern research on EOs from Chinese medicinals, the method for studying chemical penetration enhancers is often used, which fails to reflect the overall efficacy of EOs. This study clarified the property regularity of EOs from Chinese medicinals as transdermal penetration enhancers, and thereby proposed a research model which integrated the medicinal and adjuvant properties of EOs from Chinese medicinals via "component-delivery-effect" characterization route. The core concept is that constituents of EOs from Chinese medicinals and their delivery process play a key role in their external application. This research model is expected to serve as a reference for further research on EOs from Chinese medicinals for transdermal application.

Preparation technology on essential oil from Chinese materia medica as penetration enhancers / 中草药

The critical problem to be solved in the study of transdermal drug delivery system is to improve the transdermal penetration of drugs through the skin. Most essential oils from Chinese material medica, considered as a large class of penetration enhancers, possess enhanced percutaneous absorption, minor skin irritation, certain therapeutic efficacy, and a synergistic effect with transdermal drugs. However, the essential oils have poor solubility and unstable problems. Some special preparations, including patch, clathrate, gel plasters, gels, microemulsion, and so on, can increase the solubility of the essential oils, improve the effect of permeability, and can significantly boost the stability of the essential oils. In this paper, the techniques of the related preparations on essential oil from Chinese materia medica should be reviewed, and the research direction of the later stage should be discussed.

Simultaneous determination of seven constituents in Gnaphalium affine / 中成药

AIM To establish an HPLC method for the simultaneous content determination of seven constituents in Gnaphalium affine D.Don.METHODS The analysis of 80％ methanol of G.affine was performed on a 30 ℃ Atlantis (C) T3 column (4.6 mm× 250 mm,5 μm),with the mobile phase comprising of acetonitrile-formic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 288 nm.RESULTS Seven constituents showed good linear relationships within their own ranges (R2 ≥0.999 8),whose average recoveries were 98.58％-103.8％ with the RSDs of 0.88％-1.74％.CONCLUSION This accurate,stable and reproducible method can be used for the quality control of G.affine.

Chemical constituents from Illicium brevistylum and their anti-inflammatory activities / 中成药

AIM To study the chemical constituents from Illicium brevistylum A.C.Smith and to evaluate their anti-inflammatory activities.METHODS The n-BuOH fraction of 80％ ethanol extract from I.brevistylum was isolated and purified by silica,ODS and Sephadex LH-20,the structures of obtained compounds were identified by spectral data.Then their anti-inflammatory activities were screened.RESULTS Nine compounds were isolated and identified as (7S,8R)-3,3',5-trimethoxy-4',7-epoxy-8,5'-neolignan-4,9,9'-triol (1),methylabieta-8,11,13,15-tetraen-18-oate (2),majusanin A (3),pubeside C (4),(+)-lyoniresinol-3a-O-α-L-rhamnopyranoside (5),junipercomnoside D (6),lyoniside (7),nudiposide (8),rhyncoside A (9).The inhibition rates of compounds 2,6,9 on NF-κB were 47.81％,37.33％,33.37％,respectively.CONCLUSION Compounds 1,5,7-9 are isolated from genus Illicium for the first time,and compounds 2,6,9 exhibit good anti-inflammatory activities.

Chemical constituents from Illicium brevistylum and their anti-inflammatory activities / 中成药

AIM To study the chemical constituents from Illicium brevistylum A.C.Smith and to evaluate their anti-inflammatory activities.METHODS The n-BuOH fraction of 80％ ethanol extract from I.brevistylum was isolated and purified by silica,ODS and Sephadex LH-20,the structures of obtained compounds were identified by spectral data.Then their anti-inflammatory activities were screened.RESULTS Nine compounds were isolated and identified as (7S,8R)-3,3',5-trimethoxy-4',7-epoxy-8,5'-neolignan-4,9,9'-triol (1),methylabieta-8,11,13,15-tetraen-18-oate (2),majusanin A (3),pubeside C (4),(+)-lyoniresinol-3a-O-α-L-rhamnopyranoside (5),junipercomnoside D (6),lyoniside (7),nudiposide (8),rhyncoside A (9).The inhibition rates of compounds 2,6,9 on NF-κB were 47.81％,37.33％,33.37％,respectively.CONCLUSION Compounds 1,5,7-9 are isolated from genus Illicium for the first time,and compounds 2,6,9 exhibit good anti-inflammatory activities.

Correlation between Intravoxel Incoherent Motion Magnetic Resonance Imaging Derived Metrics and Serum Soluble CD40 Ligand Level in an Embolic Canine Stroke Model

OBJECTIVE: To determine the relationship between intravoxel incoherent motion (IVIM) imaging derived quantitative metrics and serum soluble CD40 ligand (sCD40L) level in an embolic canine stroke model. MATERIALS AND METHODS: A middle cerebral artery occlusion model was established in 24 beagle dogs. Experimental dogs were divided into low- and high-sCD40L group according to serum sCD40L level at 4.5 hours after establishing the model. IVIM imaging was scanned at 4.5 hours after model establishment using 10 b values ranging from 0 to 900 s/mm². Quantitative metrics diffusion coefficient (D), pseudodiffusion coefficient (D*), and perfusion fraction (f) of ischemic lesions were calculated. Quantitative metrics of ischemic lesions were normalized by contralateral hemisphere using the following formula: normalized D = D(stroke) / D(contralateral). Differences in IVIM metrics between the low- and high-sCD40L groups were compared using t test. Pearson's correlation analyses were performed to determine the relationship between IVIM metrics and serum sCD40L level. RESULTS: The high-sCD40L group showed significantly lower f and normalized f values than the low-sCD40L group (f, p 0.05). Both f and normalized f values were negatively correlated with serum sCD40L level (f, r = −0.789, p 0.05). CONCLUSION: The f value derived from IVIM imaging was negatively correlated with serum sCD40L level. f value might serve as a potential imaging biomarker to assess the formation of microvascular thrombosis in hyperacute period of ischemic stroke.

Implications of newly-added N-glycosylation mutation of hepatitis B virus s-gene in patients with coexistence of HBsAg and antiHBs / 解放军医学杂志

Objective To analyze the characteristics of newly added N-glycosylation mutation in major hydrophilic region (MHR) of HBV S gene in patients with coexistence of HBsAg and antiHBs, and reveal the generation mechanism and clinical implications of the coexistence. Methods HBV S genes from 284 patients with HBsAg+antiHBs and 314 patients with single HBsAg were amplified respectively for sequence analysis. A chronic hepatitis B (CHB) patient with HBsAg+antiHBs in MHR was found to harbor a novel double N-glycosylation mutation and selected for further study. Recombinant vectors harboring the novel mutant or control PreS/S genes were constructed and transfected in HepG2 cells respectively for phenotypic analysis, and the effects of the mutations on HBV duplication and antigenicity were investigated. Results The detection rate of MHR N-glycosylation mutation was significantly higher in HBsAg+antiHBs group than in single HBsAg group (11.3% vs. 2.9%, P<0.01, respectively). In HBsAg+antiHBs cohort, the proportion of hepatocellular carcinoma (HCC) patients accounted for 46.9%(15/32) in patients with N-glycosylation mutation at the time of testing; by contrast, the number was 22.6%(57/252) in patients with non-N-glycosylation mutation (P<0.01). N-glycosylation mutational pattern of the novel strain was s116-118TST → NST+s131-133TSM → NST concomitant with sP120 deletion+G145D mutation. The novel mutants accounted for 98.0%, 2.0% and 2.5%, respectively, of viral clones in three sequential serum samples. Mutants with single N-glycosylation mutation s130-132GTS→NSS without sP120 deletion+G145D were detected in sample 2, accounting for 17.6% of viral clones. Compared to the wild-type, the novel mutant had an increase of 31% in replication capacity, but a decrease of 99% in HBsAg level. Immunofluorescence showed that elimination of the two additional N-glycosylation mutations only partly restored HBsAg detection by antiHBs, suggesting that sP120 deletion+G145D mutation also attenuated HBsAg antigenicity. Conclusions Additional N-glycosylation mutation in MHR of HBV S gene is associated with coexisting HBsAg+antiHBs, and the two parameters together might be a better risk factor for HCC occurrence. Combination of two additional N-glycosylation mutation, sP120 deletion and sG145D mutation may co-play a role in silence of HBsAg antigenicity.

Morphological evidence of telocytes in mice aorta / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Telocytes (TCs) are a novel type of interstitial cells, which have been recently described in a large variety of cavitary and noncavitary organs. TCs have small cell bodies, and remarkably thin, long, and moniliform prolongations called telopodes (Tps). Until now, TCs have been found in various loose connective tissues surrounding the arterioles, venules, and capillaries, but as a histological cellular component, whether TCs exist in large arteries remains unexplored.</p><p><b>METHODS</b>TCs were identified by transmission electron microscope in the aortic arch of male C57BL/6 mice.</p><p><b>RESULTS</b>TCs in aortic arch had small cell bodies (length: 6.06-13.02 μm; width: 1.05-4.25 μm) with characteristics of specific long (7.74-39.05 μm), thin, and moniliform Tps; TCs distributed in the whole connective tissue layer of tunica adventitia: TCs in the innermost layer of tunica adventitia, located at the juncture between media and adventitia, with their long axes oriented parallel to the outer elastic membrane; and TCs in outer layers of tunica adventitia, were embedded among transverse and longitudinal oriented collagen fibers, forming a highly complex three-dimensional meshwork. Moreover, desmosomes were observed, serving as pathways connecting neighboring Tps. In addition, vesicles shed from the surface of TCs into the extracellular matrix, participating in some biological processes.</p><p><b>CONCLUSIONS</b>TCs in aorta arch are a newly recognized complement distinct from other interstitial cells in large arteries, such as fibroblasts. And further biologically functional correlations need to be elucidated.</p>

Comparative Analysis of Telomerase Activity in CD117⁺ CD34⁺ Cardiac Telocytes with Bone Mesenchymal Stem Cells, Cardiac Fibroblasts and Cardiomyocytes / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>This study characterized the cardiac telocyte (TC) population both in vivo and in vitro, and investigated its telomerase activity related to mitosis.</p><p><b>METHODS</b>Using transmission electron microscopy and a phase contrast microscope, the typical morphological features of cardiac TCs were observed; by targeting the cell surface proteins CD117 and CD34, CD117 + CD34 + cardiac TCs were sorted via flow cytometry and validated by immunofluorescence based on the primary cell culture. Then the optimized basal nutrient medium for selected population was examined with the cell counting kit 8. Under this conditioned medium, the process of cell division was captured, and the telomerase activity of CD117 + CD34 + cardiac TCs was detected in comparison with bone mesenchymal stem cells (BMSCs), cardiac fibroblasts (CFBs), cardiomyocytes (CMs).</p><p><b>RESULTS</b>Cardiac TCs projected characteristic telopodes with thin segments (podomers) in alternation with dilation (podoms). In addition, 64% of the primary cultured cardiac TCs were composed of CD117 + CD34 + cardiac TCs; which was verified by immunofluorescence. In a live cell imaging system, CD117 + CD34 + cardiac TCs were observed to enter into cell division in a short time, followed by an significant invagination forming across the middle of the cell body. Using a real-time quantitative telomeric-repeat amplification assay, the telomerase concentration in CD117 + CD34 + cardiac TCs was obviously lower than in BMSCs and CFBs, and significantly higher than in CMs.</p><p><b>CONCLUSIONS</b>Cardiac TCs represent a unique cell population and CD117 + CD34 + cardiac TCs have relative low telomerase activity that differs from BMSCs, CFBs and CMs and thus they might play an important role in maintaining cardiac homeostasis.</p>

Effect of Morinda officinalis capsule on osteoporosis in ovariectomized rats / 中国天然药物

AIM@#To explore the therapeutic effects of Morinda officinalis capsules (MOP) on osteoporosis in ovariectomized rats.@*METHODS@#Six-month-old female Sprague-Dawley rats were induced for postmenopausal osteoporosis (PMOP) by bilateral ovariectomy and divided into seven groups as follows: sham-operated group, ovariectomized (OVX) control group, OVX treated with xianlinggubao (XLGB) (270 mg·kg⁻¹·d⁻¹), OVX treated with alendronate sodium (ALN) (3 mg·kg⁻¹·d⁻¹), and OVX treated with Morinda officinalis capsule (MOP) of graded doses (90, 270 and 810 mg·kg⁻¹·d⁻¹) groups. Oral treatments were administered daily on the 4(th) week after ovariectomy and lasted for 12 weeks. The bone mineral density was evaluated by dual-energy X-ray absorptiometry. The tartrate-resistant acid phosphatase (TRAP), alkaline phosphatase (AKP), and osteocalcin (OC) levels in the serum and plasma were determined by standard colorimetric and enzyme immunoassays methods. Bone biomechanical properties and morphological parameters were analyzed by three-point bending test and histomorphometry respectively.@*RESULTS@#Morinda officinalis capsules at all doses were able to significantly prevent the OVX-induced loss of bone mass due to diminishing serum AKP and TRAP levels while elevating OC level in the plasma. Morinda officinalis capsules also enhanced the bone strength and prevented the deterioration of trabecular microarchitecture.@*CONCLUSION@#Morinda officinalis capsules possess potent anti-osteoporotic activity in OVX rats which could be an effective treatment for postmenopausal osteoporosis.

Multimodal magnetic resonance imaging for assessing lacunar infarction after proximal middle cerebral artery occlusion in a canine model / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>A new lacunar infarction model was recently established in beagle dogs through proximal middle cerebral artery (MCA) occlusion by thrombus. This study aimed to characterize the model by multimodal magnetic resonance imaging (MRI) and to investigate its potential role for the future stroke research.</p><p><b>METHODS</b>The left proximal MCA was embolized with an autologous thrombus in six beagles. Diffusion-weighted imaging (DWI) and T2-weighted imaging (T2WI) were performed every half hour during the first six hours after occlusion, followed by three time points at 12 hours, 24 hours, and one week. Perfusion-weighted imaging (PWI) and magnetic resonance angiography (MRA) were carried out at six hours, 24 hours and one week. The PWI-DWI mismatch ratio was defined as (PWI-DWI)/DWI ischemic volume.</p><p><b>RESULTS</b>Lacunar infarcts induced by MCA occlusion were located in the left caudate nucleus and internal capsule. All the lesions could be detected within two hours by DWI. Lesion volume on DWI increased in a time dependent manner, from (87.19 ± 67.16) mm(3) at one hour up to (368.98 ± 217.05) mm(3) at 24 hours (P = 0.009), while that on PWI gradually decreased from (7315.00 ± 2054.38) mm(3) at six hours to (4900.33 ± 1319.71) mm(3) at 24 hours and (3334.33 ± 1195.11) mm(3) at one week (P = 0.002). The mismatch ratio was 41.93 ± 22.75 at six hours after ischemia, showing "extensive mismatch", and decreased to 18.10 ± 13.74 at 24 hours (P = 0.002). No MCA recanalization was observed within 24 hours after MCA occlusion.</p><p><b>CONCLUSIONS</b>Lacunar infarction induced by proximal MCA occlusion could be detected early by DWI and was characterized by extensive PWI-DWI mismatch. Multimodal MRI is useful to demonstrate the natural evolution of PWI-DWI mismatch. This ischemic model could be further used for investigating early thrombolysis in lacunar stroke showing extensive mismatch.</p>

Study on preparation of quercetin nanostructured lipid carriers and their physicochemical properties / 中国中药杂志

<p><b>OBJECTIVE</b>To prepare quercetin nanostuctured lipid carriers (QT-NLC), and detect their physicochemical properties.</p><p><b>METHOD</b>QT-NLC was prepared by emulsification ultrasonic dispersion method, and the optimum prescription was screened out by orthogonal design. Transmission electron microscope was used to observe QT-NLC morphology. Granulometer was applied to determine zeta potential, particle size and distribution. DSC was adopted for phase analysis. Centrifugal ultra-filtration method was used to determine entrapment efficiency. Dialysis method was adopted to detect drug release in vitro of preparations.</p><p><b>RESULT</b>QT-NLC prepared under optimum conditions was mostly spherical grains, with the average particle size of (175 +/- 25) nm, which were distributed evenly, and zeta potential was (-23 +/- 0.3) mV. DSC results indicated that the drug was dispersed in nano-particles in a non-crystalline state, with an entrapment efficiency of (95.43 +/- 0.23)% and a drug-loading capacity of (2.38 +/- 0.24)%. The in vitro drug release was 32.2% in 2 hours, which was followed by a sustained release.</p><p><b>CONCLUSION</b>Emulsification ultrasonic dispersion method is applicable for preparing QT-NLC, as nano-particles are distributed evenly, with good reliability. This processing technology is safe, reliable and highly reproducible.</p>

Detection of EGFR and K-ras mutations in non-small cell lung cancer using cytological specimens / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To validate the feasibility for detecting EGFR and k-ras mutations using cytological specimens.</p><p><b>METHODS</b>Cytological specimens including fine-needle aspiration (FNA), pleural effusion (PLE) and fiberoptic bronchoscopic (FOB) brushing were collected from patients with non-small cell lung cancer (NSCLC ) from January 2011 to July 2011 at the Department of Cytology, Cancer Hospital, Chinese Academy of Medical Sciences (CHCAMS). Polymerase chain reaction (PCR) was carried out to amplify EGFR exons 18-21 and k-ras codons 12-13, and then the PCR products sequencing and analysis were performed.</p><p><b>RESULTS</b>Fifty cytological specimens were collected including 19 cases of FOB, 9 cases of FNA, 22 cases of PLE. Of them DNA was successfully extracted in 43 cases, and specific PCR amplification products sequencing were performed in 42 cases. EGFR mutations were detected in 14 of 42 specimens (33.3%), the frequencies of EGFR mutations in exons 19, 20 and 21 were 16.7% (7/42), 4.8% (2/42) and 11.9% (5/42), respectively, and no mutation was found in exon 18. Higher frequencies of EGFR mutations were detected in exons 19 and 21 (85.7%). Mutations were identified in 38.7% (12/31) cases of adenocarcinoma. K-ras mutations were found in 2 of 42 specimens (4.8%). EGFR and K-ras mutations were not found in the same case.</p><p><b>CONCLUSIONS</b>Cytological specimens are feasible for detecting EGFR and K-ras mutation. This is especially beneficial in patients in whom histological materials can not be obtained.</p>

Expression of EGFR, survivn and hnRNPA2/B1 in bronchial brushing cytology specimens and their significance in diagnosis of lung cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the expression of EGFR, survivn and hnRNPA2/B1 proteins in ThinPrep bronchial brushing cytology specimens and their significance in diagnosis of lung cancer.</p><p><b>METHODS</b>The protein expression of EGFR, survivn and hnRNPA2/B1 was detected by immunocytochemistry (ICC) in the specimens from 110 cases of non-small cell lung cancer (NSCLC), 32 cases of small cell lung cancer (SCLC) and 37 cases of non-neoplastic lung lesions. The relationship between EGFR, survivn and hnRNPA2/B1 protein expression and clinical characteristics of the patients was analyzed using SPSS 16.0 software.</p><p><b>RESULTS</b>(1) Positive expression was observed in 81.1% of NSCLC for EGFR, 66.2% for survivn, 90.9% for hnRNPA2/B1, significantly higher in NSCLC than in the control specimens (P = 0.000, P = 0.000, P = 0.010). The positive expression rate of hnRNPA2/B1 in SCLC was 92.3%, significantly higher than that in the control specimens (P = 0.021). (2) The expression of EGFR was associated with differentiation (P = 0.003), clinical stage (P = 0.023) and lymph node metastasis (P = 0.038), but was not associated with gender, age and histological types. The survivn expression was not related with the above mentioned clinicopathological features (P>0.05). Expression of hnRNPA2/B1 was associated with clinical stage (P = 0.017), but not associated with gender, age, histological type, differentiation and lymph node metastasis. (3) There was a significant difference between the co-expression of EGFR and survivn in NSCLC (98.0%) and benign conditions (2.0%, P = 0.000), also a significant difference between the negative expression of both EGFR and survivn in NSCLC (38.2%) and nonneoplastic lesions (61.8%, P = 0.000).</p><p><b>CONCLUSIONS</b>Analysis of EGFR, survivn and hnRNPA2/B1 expression may be an useful adjunct method to stratify controversial cases. The positive expression of EGFR might be associated with invasion, progression and poor prognosis of NSCLC.</p>

Relationship between prognosis of nasal inverted papilloma and human papillomavirus types / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To detected HPV human papillomavirus DNA and HPV types in NIP nasal inverted papilloma, to inquire into the infection of HPV in the pathogenesis of NIP, and the relationship of HPV with the prognosis of NIP.</p><p><b>METHODS</b>Twenty-eight cases of NIP were divided into 3 groups: no recurrence group (group 1), recurrence group (group 2), squamous cell carcinoma (SCC) arising in NIP group (malignancy group, group 3). Ten cases of benign nasal polyps were used as control group. HPV-DNA types of 6, 11, 16, 18 and general type were detected by polymerase chain reaction (PCR).</p><p><b>RESULTS</b>Total positive rate of HPV in NIP was 75% (21/28). The positive rate of group 1 was 42% (5/12), all with single and low risk HPV type infection (4 with HPV6 and 1 with HPV11). Thirteen cases of recurrence and 3 cases of malignancy were discovered to have HPV-DNA, in group 2, the majority were HPV6 and HPV11, 4 cases with double infection;in group 3, the majority were HPV16 and HPV18, and 2 cases with double infection.</p><p><b>CONCLUSIONS</b>The occurrence of NIP was related with HPV infection. To detected HPV and its subtypes can show the cases easily to have recurrence or malignant change. For the cases with HPV positive, double infection and infected with high risk types should be closely followed-up.</p>